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Sample records for amperometric immunosensor based

  1. An amperometric chloramphenicol immunosensor based on cadmium sulfide nanoparticles modified-dendrimer bonded conducting polymer.

    Science.gov (United States)

    Kim, Dong-Min; Rahman, Md Aminur; Do, Minh Hien; Ban, Changill; Shim, Yoon-Bo

    2010-03-15

    An amperometric chloramphenicol (CAP) immunosensor was fabricated by covalently immobilizing anti-chloramphenicol acetyl transferase (anti-CAT) antibody on cadmium sulfide nanoparticles (CdS) modified-dendrimer that was bonded to the conducting polymer (poly 5, 2': 5', 2''-terthiophene-3'-carboxyl acid (poly-TTCA)) layer. The AuNPs, dendrimers, and CdS nanoparticles were deposited onto the polymer layer in order to enhance the sensitivity of the sensor probes. The particle sizes were determined using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The immobilization of dendrimers, CdS, and anti-CAT were confirmed using energy disruptive spectroscopy (EDS), X-ray photoelectron spectroscopy (XPS), and quartz crystal microbalance (QCM) techniques. The detection of CAP was based on the competitive immuno-interaction between the free- and labeled-CAP for active sites of the anti-CAT. Hydrazine was used as the label for CAP, and it electrochemically catalyzed the reduction of H(2)O(2) at -0.35 V vs. Ag/AgCl. Under optimized conditions, the proposed immunosensor exhibited a linear range of CAP detection between 50 pg/mL and 950 pg/mL, and the detection limit was 45 pg/mL. The immunosensor was examined in real meat samples for the analysis of CAP.

  2. Double electrochemical covalent coupling method based on click chemistry and diazonium chemistry for the fabrication of sensitive amperometric immunosensor.

    Science.gov (United States)

    Qi, Honglan; Li, Min; Zhang, Rui; Dong, Manman; Ling, Chen

    2013-08-20

    A double electrochemical covalent coupling method based on click chemistry and diazonium chemistry for the fabrication of sensitive amperometric immunosensor was developed. As a proof-of-concept, a designed alkyne functionalized human IgG was used as a capture antibody and a HRP-labeled rabbit anti-goat IgG was used as signal antibody for the determination of the anti-human IgG using the sandwich model. The immunosensor was fabricated by electrochemically grafting a phenylazide on the surface of a glassy carbon electrode, and then, by coupling the alkyne functionalized human IgG with the phenylazide group through an electro-click chemistry in the presence of Cu(II). The amperometric measurement for the determination of the anti-human IgG was performed after the fabricated immunosensor was incubated with the target anti-human IgG and then with the HRP-labeled anti-goat IgG at -0.25V in 0.10M PBS (pH 7.0) containing 0.1mM hydroquinone and 2.0mM H2O2. The results showed that the increased current was linear with the logarithm of the concentration of the anti-human IgG in the range from 1.0×10(-10)g mL(-1) to 1.0×10(-8)g mL(-1) with a detection limit of 3×10(-11)g mL(-1). Furthermore, the feasibility of the double electrochemical covalent coupling method proposed in this work for fabricating the amperometric immunosensor array was explored. This work demonstrates that the double electrochemical covalent coupling method is a promising approach for the fabrication of the immunosensor and immunosensor array.

  3. Modeling of Amperometric Immunosensor for CMOS Integration

    Institute of Scientific and Technical Information of China (English)

    Ce Li; Haigang Yang; Shanhong Xia; Chao Bian

    2006-01-01

    A circuit model of the Amperometric immunosensor for use in the biosensor system-on-chip simulation is proposed in this paper. The model parameters are extracted with several methods and verified by MATLAB and SPICE simulation. A CMOS potentiostat circuit required for conditioning the Amperometric immunosensor is also included in the circuit model. The mean square error norm of the simulated curve against the measured one is 8.65 × 10-17. The whole circuit has been fabricated in a 0.35am CMOS process.

  4. Amperometric Immunosensor Based on a Protein A/Deposited Gold Nanocrystals Modified Electrode for Carbofuran Detection

    Directory of Open Access Journals (Sweden)

    Xia Sun

    2011-12-01

    Full Text Available In this paper, an amperometric immunosensor modified with protein A/deposited gold nanocrystals (DpAu was developed for the ultrasensitive detection of carbofuran residues. First, DpAu were electrodeposited onto the Au electrode surface to absorb protein A (PA and improve the electrode conductivity. Then PA was dropped onto the surface of DpAu film, used for binding antibody Fc fragments. Next, anti-carbofuran monoclonal antibody was immobilized on the PA modified electrode. Finally, bovine serum albumin (BSA was employed to block the possible remaining active sites avoiding any nonspecific adsorption. The fabrication procedure of the immunosensor was characterized by electrochemical impedance spectroscopy (EIS and cyclic voltammetry (CV, respectively. With the excellent electroconductivity of DpAu and the PA’s oriented immobilization of antibodies, a highly efficient immuno-reaction and detection sensitivity could be achieved. The influences of the electrodeposition time of DpAu, pH of the detection solution and incubation time on the current response of the fabricated immunosensor were investigated. Under optimized conditions, the current response was proportional to the concentration of carbofuran which ranged from 1 to 100 ng/mL and 100 ng/mL to 100 μg/mL. The detection limit was 0.1924 ng/mL. The proposed carbofuran immnuosensor exhibited high specificity, reproducibility, stability and regeneration performance, which may open a new door for ultrasensitive detection of carbofuran residues in vegetables and fruits.

  5. Amperometric immunosensor based on a protein A/deposited gold nanocrystals modified electrode for carbofuran detection.

    Science.gov (United States)

    Sun, Xia; Zhu, Ying; Wang, Xiangyou

    2011-01-01

    In this paper, an amperometric immunosensor modified with protein A/deposited gold nanocrystals (DpAu) was developed for the ultrasensitive detection of carbofuran residues. First, DpAu were electrodeposited onto the Au electrode surface to absorb protein A (PA) and improve the electrode conductivity. Then PA was dropped onto the surface of DpAu film, used for binding antibody Fc fragments. Next, anti-carbofuran monoclonal antibody was immobilized on the PA modified electrode. Finally, bovine serum albumin (BSA) was employed to block the possible remaining active sites avoiding any nonspecific adsorption. The fabrication procedure of the immunosensor was characterized by electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV), respectively. With the excellent electroconductivity of DpAu and the PA's oriented immobilization of antibodies, a highly efficient immuno-reaction and detection sensitivity could be achieved. The influences of the electrodeposition time of DpAu, pH of the detection solution and incubation time on the current response of the fabricated immunosensor were investigated. Under optimized conditions, the current response was proportional to the concentration of carbofuran which ranged from 1 to 100 ng/mL and 100 ng/mL to 100 μg/mL. The detection limit was 0.1924 ng/mL. The proposed carbofuran immnuosensor exhibited high specificity, reproducibility, stability and regeneration performance, which may open a new door for ultrasensitive detection of carbofuran residues in vegetables and fruits.

  6. Amperometric Immunosensor for Carbofuran Detection Based on MWCNTs/GS-PEI-Au and AuNPs-Antibody Conjugate

    Directory of Open Access Journals (Sweden)

    Xiangyou Wang

    2013-04-01

    Full Text Available In this paper, an amperometric immunosensor for the detection of carbofuran was developed. Firstly, multiwall carbon nanotubes (MWCNTs and graphene sheets-ethyleneimine polymer-Au (GS-PEI-Au nanocomposites were modified onto the surface of a glass carbon electrode (GCE via self-assembly. The nanocomposites can increase the surface area of the GCE to capture a large amount of antibody, as well as produce a synergistic effect in the electrochemical performance. Then the modified electrode was coated with gold nanoparticles-antibody conjugate (AuNPs-Ab and blocked with BSA. The monoclonal antibody against carbofuran was covalently immobilized on the AuNPs with glutathione as a spacer arm. The morphologies of the GS-PEI-Au nanocomposites and the fabrication process of the immunosensor were characterized by X-ray diffraction (XRD, ultraviolet and visible absorption spectroscopy (UV-vis and scanning electron microscopy (SEM, respectively. Under optimal conditions, the immunosensor showed a wide linear range, from 0.5 to 500 ng/mL, with a detection limit of 0.03 ng/mL (S/N = 3. The as-constructed immunosensor exhibited notable performance features such as high specificity, good reproducibility, acceptable stability and regeneration performance. The results are mainly due to the excellent properties of MWCNTs, GS-PEI-Au nanocomposites and the covalent immobilization of Ab with free hapten binding sites for further immunoreaction. It provides a new avenue for amperometric immunosensor fabrication.

  7. Amperometric immunosensor for carbofuran detection based on MWCNTs/GS-PEI-Au and AuNPs-antibody conjugate.

    Science.gov (United States)

    Zhu, Ying; Cao, Yaoyao; Sun, Xia; Wang, Xiangyou

    2013-04-19

    In this paper, an amperometric immunosensor for the detection of carbofuran was developed. Firstly, multiwall carbon nanotubes (MWCNTs) and graphene sheets-ethyleneimine polymer-Au (GS-PEI-Au) nanocomposites were modified onto the surface of a glass carbon electrode (GCE) via self-assembly. The nanocomposites can increase the surface area of the GCE to capture a large amount of antibody, as well as produce a synergistic effect in the electrochemical performance. Then the modified electrode was coated with gold nanoparticles-antibody conjugate (AuNPs-Ab) and blocked with BSA. The monoclonal antibody against carbofuran was covalently immobilized on the AuNPs with glutathione as a spacer arm. The morphologies of the GS-PEI-Au nanocomposites and the fabrication process of the immunosensor were characterized by X-ray diffraction (XRD), ultraviolet and visible absorption spectroscopy (UV-vis) and scanning electron microscopy (SEM), respectively. Under optimal conditions, the immunosensor showed a wide linear range, from 0.5 to 500 ng/mL, with a detection limit of 0.03 ng/mL (S/N = 3). The as-constructed immunosensor exhibited notable performance features such as high specificity, good reproducibility, acceptable stability and regeneration performance. The results are mainly due to the excellent properties of MWCNTs, GS-PEI-Au nanocomposites and the covalent immobilization of Ab with free hapten binding sites for further immunoreaction. It provides a new avenue for amperometric immunosensor fabrication.

  8. A micro amperometric immunosensor for detection of human immunoglobulin

    Institute of Scientific and Technical Information of China (English)

    XU Yuanyuan; XIA Shanhong; BIAN Chao; CHEN Shaofeng

    2006-01-01

    A novel amperometric immunosensor based on the micro electromechanical systems (MEMS) technology, using protein A and self-assembled monolayers (SAMs) for the orientation-controlled immobilization of antibodies, has been developed. Using MEMS technology, an "Au, Pt, Pt" three-microelectrode system enclosed in a SU-8 micro pool was fabricated. Employing SAMs, a monolayer of protein A was immobilized on the cysteamine modified Au electrode to achieve the orientation-controlled immobilization of the human immunoglobulin (HIgG) antibody. The immunosensor aimed at low unit cost, small dimension, high level of integration and the prospect of a biosensor system-on-a-chip. Cyclic voltammetry and chronoamperometry were conducted to characterize the immunosensor. Compared with the traditional immunosensor using bulky gold electrode or screen-printed electrode and the procedure directly binding protein A to electrode for immobilization of antibodies, it had attractive advantages, such as miniaturization, compatibility with CMOS technology, fast response (30 s), broad linear range (50-400μg/L) and low detection limit (10μg/L) for HIgG. In addition, this immunosensor was easy to be designed into micro array and to realize the simultaneously multi-parameter detection.

  9. Micro amperometric immunosensor for the detection of salmonella typhimurium

    Science.gov (United States)

    Sun, Jizhou; Xia, Shanhong; Bian, Chao; Qu, Lan

    2008-12-01

    In this paper, a micro amperometric immunosensor based on Micro-Electro-Mechanical Systems technology for the detection of Salmonella typhimurium (S. typhimurium) was constructed by immobilizing a polyclonal antibody (the bio-molecular recognition element) onto the surface of polypyrrole(PPy) /staphylococcal protein A(SPA) modified Pt electrode. Pyrrole doped with SPA was co-electropolymerized onto the working electrode surface by cyclic voltammetry in 10 minutes for orientation-controlled immobilization of salmonella capture antibodies. S. typhimurium with the concentration of 102cfu/ml could be detected by this immunosensor with a controllable and convenient manipulation to effectively modify the sensing surface more rapidly with less consumption of reagent (10µL), which showed the good property of the sensor. It is potential to develop a micro biosensor that can be used for convenient, accurate, cost-effective and real-time sensing of pathogens in food products.

  10. A sensitive label-free amperometric CEA immunosensor based on graphene-nafion nanocomposite film as an enhanced sensing platform.

    Science.gov (United States)

    Li, Yan; Yang, Wei-Kang; Fan, Man-Qi; Liu, Ao

    2011-01-01

    A novel approach to fabricate a label-free amperometric immunosensor for the detection of carcinoembryonic antigen (CEA) was described. Herein, methylene blue (MB), gold nanoparticles (AuNPs) and carcinoembryonic antibody (anti-CEA) were layer-by-layer assembled on the graphene-Nafion nanocomposite film-modified electrode by means of a self-assembling technique and the opposite-charged adsorption. Subsequently, the stepwise self-assembling procedure of the immunosensor was further characterized by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The factors influencing the performance of the resulting immunosensor were studied in detail. The developed procedure showed improved features, including larger amount and higher immunoactivity of the immobilized antibody and repeatable regeneration of the sensor, as well as direct, rapid and simple determination for the antigen without multiple separation and labeling steps. The immunosensor could detect the target protein in a range of 0.5 to 120 ng/mL with a limit of 0.17 ng/mL (at 3σ). Finally, the immunosensing system was evaluated on several clinical samples. Analytical results were found to be in satisfactory agreement with those detected by the enzyme-linked immunosorbent assay (ELISA) method, indicating that this new method was a promising alternative tool for clinical diagnosis.

  11. Amperometric immunosensor for {alpha}-fetoprotein antigen in human serum based on co-immobilizing dinuclear copper complex and gold nanoparticle doped chitosan film

    Energy Technology Data Exchange (ETDEWEB)

    Gan Ning; Meng Linghua; Wang Feng [State Key Laboratory Base of Novel Functional Materials and Preparation science, Faculty of Material Science and Chemical Engineering of Ninbo University, Ningbo, 315211 (China)], E-mail: ganning@nbu.edu.cn

    2009-09-01

    A sensitive amperometric immunosensor for {alpha}-fetoprotein (AFP), a tumor marker for the diagnosis of hepatocellular carcinoma (HCC), was constructed, The immunosensor is prepared by co-immobilizing [Cu{sub 2}(phen){sub 2}Cl{sub 2}] ({mu}-Cl){sub 2} (CuL), nano-Au/Chitosan(Chit) composite, horseradish peroxidase (HRP) and AFP antibody(anti-AFP) on a glassy carbon electrode (GCE). Firstly, CuL was irreversibly absorb on GCE electrode through {pi}-{pi} stacking interaction; then nano-Au/Chit composite was immobilized onto the electrode because of its excellent membrane-forming ability, finally HRP and anti-AFP was adsorbed onto the surface of the gold nanoparticles to construct GCE | CuL/nanoAu-chit/HRP/anti-AFP immunosensor. The preparation procedure of the electrode was characterized by electrochemical and spectroscopy method. The results showed that this immunosensor exhibited an excellent electrocatalytic response to the reduction of hydrogen peroxide (H{sub 2}O{sub 2}) without the aid of an electron mediator, offers a high-sensitivity (1710 nA {center_dot} ng{sup -1} {center_dot} ml{sup -1}) for the detection of AFP and has good correlation for detection of AFP in the range of 0.2 to 120.0 ng/ml with a detection limit of 0.05 ng/ml. The biosensor showed high selectivity as well as good stability and reproductivity.

  12. Disposable amperometric immunosensor based on layer-by-layer electro-depositing of the nanogold particles, prussian blue-modified indium tin oxide for determination of -fetoprotein

    Indian Academy of Sciences (India)

    Yan Li; Wen-Bin Liang; Li-Chao Fang; Hui Huang; Jun Deng; Jun-Song Zheng

    2009-11-01

    In this paper, a novel disposable immunosensor for the detection of -fetoprotein (AFP) based on the Indium tin oxide (ITO) modified by the sequential electro-deposition of the nanogold particles (nano-Au) and prussian blue (PB) is described. The ITO is employed to reduce the cost, instead of expensive gold electrode, glassy carbon electrode or platinum electrode. The layer-by-layer (LBL) electro-deposition of the nano-Au, PB is used for blocking the possible leakage from the substrate electrode surface and to prevent shedding of composite membrane. Under optimal conditions, the proposed immunosensor displays a broad linear response to AFP, the working range being 0.25 to 300.0 ng mL-1 with a detection limit of 0.04 ng mL-1. The studied immunosensor exhibits high sensitivity, fast analytical time and good stability. The proposed methodology is potentially attractive for clinical immunoassays.

  13. Study of the Amperometric Immunosensor Based on Gold Nanoparticles and Magnetic Nanocomposite%纳米金与磁性纳米复合物构建电流型免疫传感器的研究

    Institute of Scientific and Technical Information of China (English)

    朱宇萍; 袁若; 柴雅琴; 覃松; 苏会岚

    2012-01-01

    A new reagentless amperometric immunosensor for the detection of α-fetoprotein antibody (anti -AFP) molecule based on Nafion, Fe3O4@ thionine and gold nanoparticles was fabricated. Firstly, Fe3O4 @ thionine nanoparticles were prepared by magnetic separation method. Nafion., Fe3O4@ thionine nanoparticles were modified on glassy carbon electrode layer by layer through electrostatic adsorption, then the NH2 of thionine molecules was used to adsorbed the gold nanoparticles. Finally, with function of gold nanoparticles adsorbing AFP antibody, a reagentless amperometric AFP immunosensor with high stability and sensitivity was prepared successfully. Transmission electron microscopy and cyclic voltammetry were used to characterize the composite nanoparticles and the property of the immunosensor. Under the optimal conditions, the immunosensor had a good current response to AFP with a detection range of 0. 05 - 20 μg/L and a detection limit of 0. 03 μg/L.%制备了易于磁性分离、硫堇(Thi)包覆的四氧化三铁(Fe3O4)纳米复合物.通过静电吸附作用,将萘酚(Nafion)、Thi包覆的Fe3O4复合纳米粒子层层修饰到玻碳电极表面,再利用Thi分子中的氨基吸附纳米金,最后固载甲胎蛋白抗体,从而制得灵敏度高、稳定性好的无试剂电流型甲胎蛋白免疫传感器.实验通过透射电子显微镜(TEM)对该复合纳米粒子进行表征,并用循环伏安法考察了电极的电化学特性.结果表明,Fe3O4/Thi复合纳米粒子修饰的电极在实验过程中呈现出良好的氧化还原活性,其检测范围为0.05~20 μg/L,检出限为0.03 μg/L.

  14. Amperometric carbohydrate antigen 19-9 immunosensor based on three dimensional ordered macroporous magnetic Au film coupling direct electrochemistry of horseradish peroxidase

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Qi [College of Sciences, Nanjing Tech University, Nanjing 211816 (China); Chen, Xiaojun, E-mail: chenxj_njut@126.com [College of Sciences, Nanjing Tech University, Nanjing 211816 (China); State Key Laboratory of Materials-Oriented Chemical Engineering, Nanjing Tech University, Nanjing 210009 (China); Tang, Yin [Zhangjiagang Hospital of Traditional Chinese Medicine, Zhangjiagang 215600 (China); Ge, Lingna; Guo, Buhua [College of Sciences, Nanjing Tech University, Nanjing 211816 (China); Yao, Cheng, E-mail: yaochengnjut@163.com [College of Sciences, Nanjing Tech University, Nanjing 211816 (China)

    2014-03-01

    Highlights: • Three dimensional ordered macroporous magnetic electrode was newly used in electrochemical immunosensor. • The large surface area of macroporous magnetic electrode could improve the immobilized amount of antibody. • Au nanoparticles functionalized SBA-15 was used to immobilize enzyme labeled Ab₂ and enzyme. • Macroporous magnetic electrode and Au nanoparticles composite facilitated the direct electron transfer of enzyme. • The immunoassay avoided adding electron transfer mediator, simplifying the procedure. Abstract: A sandwich-type electrochemical immunosensor for the detection of carbohydrate antigen 19-9 (CA 19-9) antigen based on the immobilization of primary antibody (Ab₁) on three dimensional ordered macroporous magnetic (3DOMM) electrode, and the direct electrochemistry of horseradish peroxidase (HRP) that was used as both the label of secondary antibody (Ab₂) and the blocking reagent. The 3DOMM electrode was fabricated by introducing core–shell Au–SiO₂@Fe₃O₄ nanospheres onto the surface of three dimensional ordered macroporous (3DOM) Au electrode via the application of an external magnet. Au nanoparticles functionalized SBA-15 (Au@SBA-15) was conjugated to the HRP labeled secondary antibody (HRP-Ab₂) through the Au–SH or Au–NH₃⁺ interaction, and HRP was also used as the block reagent. The formation of antigen–antibody complex made the combination of Au@SBA-15 and 3DOMM exhibit remarkable synergistic effects for accelerating direct electron transfer (DET) between HRP and the electrode. Under the optimal conditions, the DET current signal increased proportionally to CA 19-9 concentration in the range of 0.05 to 15.65 U mL⁻¹ with a detection limit of 0.01 U mL⁻¹. Moreover, the immunosensor showed high selectivity, good stability, satisfactory reproducibility and regeneration. Importantly, the developed method was used to assay clinical serum specimens, achieving a good relation with those obtained from

  15. Ultrasensitive amperometric immunosensor for PSA detection based on Cu2O@CeO2-Au nanocomposites as integrated triple signal amplification strategy.

    Science.gov (United States)

    Li, Faying; Li, Yueyun; Feng, Jinhui; Dong, Yunhui; Wang, Ping; Chen, Lei; Chen, Zhiwei; Liu, Hui; Wei, Qin

    2017-01-15

    In this work, a novel label-free electrochemical immunosensor was developed for the quantitative detection of prostate specific antigen (PSA). To this end, the amino functionalized cuprous oxide @ ceric dioxide (Cu2O@CeO2-NH2) core-shell nanocomposites were prepared to bond gold nanoparticles (Au NPs) by constructing stable Au-N bond between Au NPs and -NH2. Because the synergetic effect presents in Cu2O@CeO2 core-shell loaded with Au NPs (Cu2O@CeO2-Au), it shows better electrocatalytic activity towards the reduction of hydrogen peroxide (H2O2) than single Cu2O, Au NPs and Cu2O@CeO2. Featured by large specific surface area, good biocompatibility and good electrochemical properties which can greatly improve the electronic transmission rate, Cu2O@CeO2-Au was used as transducing materials to achieve efficiently capture antibodies and triple signal amplification of the proposed immunosensor. Under the optimal conditions, the proposed immunosensor exhibited a wide linear range from 0.1pg/mL to 100ng/mL with a low detection limit of 0.03pg/mL (S/N=3). Furthermore, the proposed label-free immunosensor has been used to determine PSA in human serum with satisfactory results. Meanwhile, it displayed good reproducibility, acceptable selectivity, and long-term stability, which had promising application in bioassay analysis.

  16. Amperometric immunosensor for rapid detection of Mycobacterium tuberculosis

    Science.gov (United States)

    Hiraiwa, Morgan; Kim, Jong-Hoon; Lee, Hyun-Boo; Inoue, Shinnosuke; Becker, Annie L.; Weigel, Kris M.; Cangelosi, Gerard A.; Lee, Kyong-Hoon; Chung, Jae-Hyun

    2015-05-01

    Tuberculosis (TB) has been a major public health problem, which can be better controlled by using accurate and rapid diagnosis in low-resource settings. A simple, portable, and sensitive detection method is required for point-of-care (POC) settings. This paper studies an amperometric biosensor using a microtip immunoassay for a rapid and low-cost detection of Mycobacterium tuberculosis (MTB) in sputum. MTB in sputum is specifically captured on the functionalized microtip surface and detected by electric current. According to the numerical study, the current signal on the microtip surface is linearly changed with increasing immersion depth. Using a reference microtip, the immersion depth is compensated for a sensing microtip. On the microtip surface, target bacteria are concentrated and organized by a coffee-ring effect, which amplifies the electric current. To enhance the signal-to-noise ratio, both the sample processing and rinsing steps are presented with the use of deionized water as a medium for the amperometric measurement. When applied to cultured MTB cells spiked into human sputum, the detection limit was 100 CFU mL-1, comparable to a more labor-intensive fluorescence detection method reported previously.

  17. 基于一维DNA组装磁性纳米探针的夹心型安培免疫传感器研究%A Sandwich Amperometric Immunosensor Based on One-dimensional Assembly of Magnetic DNA Nano-probe

    Institute of Scientific and Technical Information of China (English)

    巫远招; 干宁; 胡富陶; 李天华; 曹玉廷; 郑磊

    2011-01-01

    采用Fe3O4(核)/ZrO2(壳)纳米磁珠(ZMPs)标记待测物识别抗体,并用HRP酶封闭和DNA链接,建立了一类新型的“珠链状”一维磁性纳米探针制备方法.将甲胎蛋白(AFP)一抗固定于纳米金修饰的玻碳电极表面,构建了免疫电极(GCE | AFP Ab1).基于该电极和上述合成探针,通过双抗体夹心法测定免疫产物上HRP酶对过氧化脲(CP)氧化对苯二酚反应的催化电流,研制了一类基于一维纳米结构组装的夹心型安培免疫传感器.研究表明:此一维纳米结构探针不仅大大增加了酶在电极表面的富集量,成倍扩增了催化电流,显著提高了传感器的灵敏度,而且易于通过外磁场与背景液可控分离,简化了分析步骤,并提高了结果的重复性.此传感器对AFP检测的线性范围为0.01~25tg/L;检出限达4 ng/L(3σ),并被用于人血清中痕量AFP的测定,结果满意.%A new type of one-dimensional magnetic nanoprobes was fabricated based on a DNA chain fixed with a number of Fe3O4 (core)/ZrO2 (shell) nano-magnetic beads (ZMPs) labeled with secondary antibody. The a-fetoprotein (AFP) was analyzed by using probes DNA/(ZMPs-HRP-AFP Ab2)n and immobilizing a-fetoprotein antibody (AFP Ab,) onto the glassy carbon electrode modified by nano-gold. The sandwich-type immunocomplex could be formed between the immobilized AFP Abi on the GCE and the probes DNA/( ZMPs-HRP-AFP Ab2) n and the carried HRP could catalyze the electrochemical reduction of carbamide peroxide (CP) with the help of hydroquinone (HQ). The novel one-dimensional nano structure assemble and sandwich-type amperometric immunosensor was developed. Enhanced sensitivity was achieved by introducing more HRP-AFP Ab2 onto the electrode surface through the pearl chain structure. Under optimal conditions, the proposed method could respond to 4 ng/L AFP with a linear calibration range from 0.01 to 25 ug/L. The immunosensor was employed to determine AFP in serum samples and the results

  18. Scalable fabrication of immunosensors based on carbon nanotube polymer composites

    Energy Technology Data Exchange (ETDEWEB)

    Mendoza, Ernest; Gonzalez-Guerrero, Ana B [Institut Catala de Nanotecnologia, Campus Universitat Autonoma de Barcelona, 08193 Bellaterra (Spain); Orozco, Jahir; Jimenez-Jorquera, Cecilia; Fernandez-Sanchez, Cesar [Instituto de Microelectronica de Barcelona, CNM-IMB (CSIC), Campus Universitat Autonoma de Barcelona, 08193 Bellaterra (Spain); Calle, Ana; Lechuga, Laura M [Instituto de Microelectronica de Madrid, CNM-IMM (CSIC), Isaac Newton 8, 28760 Tres Cantos, Madrid (Spain)], E-mail: Ernest.Mendoza.icn@uab.es

    2008-02-20

    In this work we present the fabrication and characterization of immunosensors based on polystyrene (PS)-multiwalled carbon nanotube (MWCNT) composites. The electrochemical properties of the sensors have been investigated and show that the surface area is increased upon addition of the MWCNT-PS layer. Furthermore, a plasma activation process is used to partially remove the PS and expose the MWCNTs. This results in a huge increase in the electrochemical area and opens up the possibility of binding biomolecules to the MWCNT wall. The MWCNTs have been functionalized covalently with a model antibody (rabbit IgG). The biosensors have been tested using amperometric techniques and show detection limits comparable to standard techniques such as ELISA.

  19. Disposable Amperometric Immunosensor for the Determination of Human P53 Protein in Cell Lysates Using Magnetic Micro-Carriers

    Science.gov (United States)

    Pedrero, María; Manuel de Villena, F. Javier; Muñoz-San Martín, Cristina; Campuzano, Susana; Garranzo-Asensio, María; Barderas, Rodrigo; Pingarrón, José M.

    2016-01-01

    An amperometric magnetoimmunosensor for the determination of human p53 protein is described in this work using a sandwich configuration involving the covalent immobilization of a specific capture antibody onto activated carboxylic-modified magnetic beads (HOOC-MBs) and incubation of the modified MBs with a mixture of the target protein and horseradish peroxidase-labeled antibody (HRP-anti-p53). The resulting modified MBs are captured by a magnet placed under the surface of a disposable carbon screen-printed electrode (SPCE) and the amperometric responses are measured at −0.20 V (vs. an Ag pseudo-reference electrode), upon addition of hydroquinone (HQ) as a redox mediator and H2O2 as the enzyme substrate. The magnetoimmunosensing platform was successfully applied for the detection of p53 protein in different cell lysates without any matrix effect after a simple sample dilution. The results correlated accurately with those provided by a commercial ELISA kit, thus confirming the immunosensor as an attractive alternative for rapid and simple determination of this protein using portable and affordable instrumentation. PMID:27879639

  20. Disposable Amperometric Immunosensor for the Determination of Human P53 Protein in Cell Lysates Using Magnetic Micro-Carriers

    Directory of Open Access Journals (Sweden)

    María Pedrero

    2016-11-01

    Full Text Available An amperometric magnetoimmunosensor for the determination of human p53 protein is described in this work using a sandwich configuration involving the covalent immobilization of a specific capture antibody onto activated carboxylic-modified magnetic beads (HOOC-MBs and incubation of the modified MBs with a mixture of the target protein and horseradish peroxidase-labeled antibody (HRP-anti-p53. The resulting modified MBs are captured by a magnet placed under the surface of a disposable carbon screen-printed electrode (SPCE and the amperometric responses are measured at −0.20 V (vs. an Ag pseudo-reference electrode, upon addition of hydroquinone (HQ as a redox mediator and H2O2 as the enzyme substrate. The magnetoimmunosensing platform was successfully applied for the detection of p53 protein in different cell lysates without any matrix effect after a simple sample dilution. The results correlated accurately with those provided by a commercial ELISA kit, thus confirming the immunosensor as an attractive alternative for rapid and simple determination of this protein using portable and affordable instrumentation.

  1. Sensitive electrochemical immunosensor based on three-dimensional nanostructure gold electrode

    Directory of Open Access Journals (Sweden)

    Zhong G

    2015-03-01

    Full Text Available Guangxian Zhong,1,2,* Ruilong Lan,3,* Wenxin Zhang,1,4 Feihuan Fu,5 Yiming Sun,1,4 Huaping Peng,1,4 Tianbin Chen,3 Yishan Cai,6 Ailin Liu,1,4 Jianhua Lin,2 Xinhua Lin1,4 1Department of Pharmaceutical Analysis, Faculty of Pharmacy, Fujian Medical University, 2Department of Orthopaedics, 3The Centralab, First Affiliated Hospital of Fujian Medical University, 4Nano Medical Technology Research Institute, Fujian Medical University, Fuzhou, 5Department of Endocrinology, The County Hospital of Anxi, Anxi, 6Fujian International Travel Healthcare Center, Fujian Entry-Exit Inspection and Quarantine Bureau, Fuzhou, People’s Republic of China *These authors contributed equally to this work Abstract: A sensitive electrochemical immunosensor was developed for detection of alpha-fetoprotein (AFP based on a three-dimensional nanostructure gold electrode using a facile, rapid, “green” square-wave oxidation-reduction cycle technique. The resulting three-dimensional gold nanocomposites were characterized by scanning electron microscopy and cyclic voltammetry. A “sandwich-type” detection strategy using an electrochemical immunosensor was employed. Under optimal conditions, a good linear relationship between the current response signal and the AFP concentrations was observed in the range of 10–50 ng/mL with a detection limit of 3 pg/mL. This new immunosensor showed a fast amperometric response and high sensitivity and selectivity. It was successfully used to determine AFP in a human serum sample with a relative standard deviation of <5% (n=5. The proposed immunosensor represents a significant step toward practical application in clinical diagnosis and monitoring of prognosis. Keywords: electrochemical immunosensors, three-dimensional nanostructure gold electrode, square-wave oxidation-reduction cycle, alpha-fetoprotein 

  2. Nanostructured conducting polymer based reagentless capacitive immunosensor.

    Science.gov (United States)

    Bandodkar, Amay Jairaj; Dhand, Chetna; Arya, Sunil K; Pandey, M K; Malhotra, Bansi D

    2010-02-01

    Nanostructured polyaniline (PANI) film electrophoretically fabricated onto indium-tin-oxide (ITO) coated glass plate has been utilized for development of an immunosensor based on capacitance change of a parallel plate capacitor (PPC) by covalently immobilizing anti-human IgG (Anti-HIgG) using N-ethyl-N'-(3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide chemistry. These fabricated PANI/ITO and Anti-HIgG/PANI/ITO plates have been characterized using scanning electron microscopy, cyclic voltammetry, differential pulse voltammetry and Fourier transform infra-red studies. The capacitance measurements indicate that dielectric medium of this biologically modified PPC (Anti-HIgG/PANI/ITO) is sensitive to HIgG in 5 - 5 x 10(5) ng mL(-1) range and has lower detection limit of 1.87 ng mL(-1). The observed results reveal that this Anti-HIgG modified PPC can be used as a robust, easy-to-use, reagentless, sensitive and selective immunosensor for estimation of human IgG.

  3. Recent Trends in Field-Effect Transistors-Based Immunosensors

    Directory of Open Access Journals (Sweden)

    Ana Carolina Mazarin de Moraes

    2016-10-01

    Full Text Available Immunosensors are analytical platforms that detect specific antigen-antibody interactions and play an important role in a wide range of applications in biomedical clinical diagnosis, food safety, and monitoring contaminants in the environment. Field-effect transistors (FET immunosensors have been developed as promising alternatives to conventional immunoassays, which require complicated processes and long-time data acquisition. The electrical signal of FET-based immunosensors is generated as a result of the antigen-antibody conjugation. FET biosensors present real-time and rapid response, require small sample volume, and exhibit higher sensitivity and selectivity. This review brings an overview on the recent literature of FET-based immunosensors, highlighting a diversity of nanomaterials modified with specific receptors as immunosensing platforms for the ultrasensitive detection of various biomolecules.

  4. Current Trends in Nanomaterial-Based Amperometric Biosensors

    Directory of Open Access Journals (Sweden)

    Akhtar Hayat

    2014-12-01

    Full Text Available The last decade has witnessed an intensive research effort in the field of electrochemical sensors, with a particular focus on the design of amperometric biosensors for diverse analytical applications. In this context, nanomaterial integration in the construction of amperometric biosensors may constitute one of the most exciting approaches. The attractive properties of nanomaterials have paved the way for the design of a wide variety of biosensors based on various electrochemical detection methods to enhance the analytical characteristics. However, most of these nanostructured materials are not explored in the design of amperometric biosensors. This review aims to provide insight into the diverse properties of nanomaterials that can be possibly explored in the construction of amperometric biosensors.

  5. A carbon nanotube-based high-sensitivity electrochemical immunosensor for rapid and portable detection of clenbuterol.

    Science.gov (United States)

    Liu, Gang; Chen, Haode; Peng, Hongzhen; Song, Shiping; Gao, Jimin; Lu, Jianxin; Ding, Min; Li, Lanying; Ren, Shuzhen; Zou, Ziying; Fan, Chunhai

    2011-10-15

    Carbon nanotubes have shown their unique advantages of mechanical, chemical and electronic properties in bioanalysis. We herein report a new method to efficiently and reproducibly prepare multi-walled carbon nanotubes (MWNTs)-protein sensing layers for electrochemical immunosensors. This method employs centrifugation to prepare a conjugate of MWNTs and goat anti mouse-immunoglobulin G (IgG) (secondary antibody). The conjugates were then deposited on screen-printed electrodes to form a nanostructured layer (MWNT-I layer). CLB monoclonal antibody was assembled through its binding to the secondary antibody. The MWNT-I layer-based electrodes were used for rapid and sensitive amperometric immunosensing detection of clenbuterol (CLB) in swine urine samples. Horseradish peroxidase-coupled CLB (CLB-HRP) competed with free CLB in the samples to bind the monoclonal antibody. It has shown significantly higher sensitivity and better reproducibility than the chemical conjugation method. This MWNT-based immunosensor is highly sensitive, leading to a limit of detection of 0.1 ng/mL within a rapid assay time of 16 min. Its sensitivity is at least 1 order of magnitude higher than that of a normal immunosensor (without MWNTs). The sensing device is portable with disposable screen-printed electrode, satisfactorily meeting the requirements for field detection of food security-related species.

  6. Numerical model based on amperometric measurements

    OpenAIRE

    Daungruthai Jarukanont; Imelda Bonifas Arredondo; Ricardo Femat; Garcia, Martin E.

    2015-01-01

    Chromaffin cells release catecholamines by exocytosis, a process that includes vesicle docking, priming and fusion. Although all these steps have been intensively studied, some aspects of their mechanisms, particularly those regarding vesicle transport to the active sites situated at the membrane, are still unclear. In this work, we show that it is possible to extract information on vesicle motion in Chromaffin cells from the combination of Langevin simulations and amperometric measurements. ...

  7. Nanomaterial-Based Electrochemical Immunosensors for Clinically Significant Biomarkers

    Directory of Open Access Journals (Sweden)

    Niina J. Ronkainen

    2014-06-01

    Full Text Available Nanotechnology has played a crucial role in the development of biosensors over the past decade. The development, testing, optimization, and validation of new biosensors has become a highly interdisciplinary effort involving experts in chemistry, biology, physics, engineering, and medicine. The sensitivity, the specificity and the reproducibility of biosensors have improved tremendously as a result of incorporating nanomaterials in their design. In general, nanomaterials-based electrochemical immunosensors amplify the sensitivity by facilitating greater loading of the larger sensing surface with biorecognition molecules as well as improving the electrochemical properties of the transducer. The most common types of nanomaterials and their properties will be described. In addition, the utilization of nanomaterials in immunosensors for biomarker detection will be discussed since these biosensors have enormous potential for a myriad of clinical uses. Electrochemical immunosensors provide a specific and simple analytical alternative as evidenced by their brief analysis times, inexpensive instrumentation, lower assay cost as well as good portability and amenability to miniaturization. The role nanomaterials play in biosensors, their ability to improve detection capabilities in low concentration analytes yielding clinically useful data and their impact on other biosensor performance properties will be discussed. Finally, the most common types of electroanalytical detection methods will be briefly touched upon.

  8. Construcción de un Inmunosensor Amperométrico utilizando Apirasa de Solanum tuberosum para la Detección de Esquistosomiasis Construction of an Amperometric Immunosensor using Solanum tuberosum potato Apyrase for the Detection of Schistosomiasis

    Directory of Open Access Journals (Sweden)

    Ninoska I Bojorge Ramírez

    2009-01-01

    Full Text Available Este trabajo presenta el desarrollo de un inmunosensor amperométrico composito a base de una proteína de origen vegetal para la determinación de antí-anticuerpo de S. mansoni. El inmunosensor consiste en una matriz electródica rígida construida con grafito y resina epoxi en la que se inmoviliza covalentemente la Apirasa. La estrategia de inmovilización se basa en silanización de la matriz compósita con 3-aminopropilsilano (3-APTES y su activación mediante glutaraldehído. La inmovilización fue monitoreada mediante voltaamperometría cíclica en Fe(CN6(4-/Fe(CN6³, obteniéndose una respuesta cuasi-reversible del inmunosensor y muy estable en el tiempo. La inmovilización también fue caracterizada mediante ángulos de contactos y microscopía electrónica de barrido. Estas cualidades son aprovechables para la determinación clínica de esquistosomiasis dada la rapidez y sensibilidad de respuesta.This paper presents the development of an amperometric composites inmunosensor based on a vegetable protein for the determination of anti-Ac S. mansoni. The inmunosensor consists of a rigid matrix built with graphite and epoxy resin in which immobilizes covalently the Apyrase. The strategy of immobilization is based in silanization of the matrix composites with 3-Aminopropyltriethoxysilane (3-APTES and its activation by glutaraldehyde. The detection was monitored by cyclic voltammetry Fe(CN6(4-/ Fe(CN6³, obtaining a response quasi-reversible of the immunosensor and very stable over time. The immobilization was also characterized by angles contacts and scanning electron microscopy. These qualities are useful for determination clinical of schistosomiasis given the speed of response and sensitivity.

  9. Amplification strategy based on gold nanoparticle-decorated carbon nanotubes for neomycin immunosensors.

    Science.gov (United States)

    Zhu, Ye; Son, Jung Ik; Shim, Yoon-Bo

    2010-11-15

    A novel amperometric immunosensor with an enhanced sensitivity for the detection of neomycin (Neo) was prepared by covalently immobilizing a monoclonal Neo antibody onto a new conducting polymer, poly-[2,5-di-(2-thienyl)-1H-pyrrole-1-(p-benzoic acid)] (pDPB), as a sensor probe. The probe was used to detect Neo in a sandwich-type approach, where the secondary antibody was attached to gold nanoparticle-decorated multi-wall carbon nanotubes labeled with hydrazine (Hyd-MWCNT(AuNP)-Ab(2)). Hydrazine on the conjugate served as a catalyst for the reduction of hydrogen peroxide, and the catalytic current was monitored at -0.45 V vs. Ag/AgCl. The performance of the immunosensor with and without AuNPs on the probe and the conjugate was compared. The parameters affecting the immunosensor response in terms of antibody dilution ratio, incubation time, pH, applied potential, and temperature were optimized. A linear dynamic range for Neo analysis was obtained between 10 ng/mL and 250 ng/mL with a detection limit of 6.76 ± 0.17 ng/mL. The proposed immunosensor was successfully applied to detect Neo content in real meat samples.

  10. An Effective Amperometric Biosensor Based on Gold Nanoelectrode Arrays

    Directory of Open Access Journals (Sweden)

    Zhu Yingchun

    2008-01-01

    Full Text Available Abstract A sensitive amperometric biosensor based on gold nanoelectrode array (NEA was investigated. The gold nanoelectrode array was fabricated by template-assisted electrodeposition on general electrodes, which shows an ordered well-defined 3D structure of nanowires. The sensitivity of the gold NEA to hydrogen peroxide is 37 times higher than that of the conventional electrode. The linear range of the platinum NEA toward H2O2is from 1 × 10−6to 1 × 10−2 M, covering four orders of magnitudes with detection limit of 1 × 10−7 M and a single noise ratio (S/N of four. The enzyme electrode exhibits an excellent response performance to glucose with linear range from 1 × 10−5to 1 × 10−2 M and a fast response time within 8 s. The Michaelis–Menten constantkm and the maximum current densityi maxof the enzyme electrode were 4.97 mM and 84.60 μA cm−2, respectively. This special nanoelectrode may find potential application in other biosensors based on amperometric signals.

  11. Novel amperometric glucose biosensor based on MXene nanocomposite

    KAUST Repository

    Rakhi, R. B.

    2016-11-10

    A biosensor platform based on Au/MXene nanocomposite for sensitive enzymatic glucose detection is reported. The biosensor leverages the unique electrocatalytic properties and synergistic effects between Au nanoparticles and MXene sheets. An amperometric glucose biosensor is fabricated by the immobilization of glucose oxidase (GOx) enzyme on Nafion solubilized Au/ MXene nanocomposite over glassy carbon electrode (GCE). The biomediated Au nanoparticles play a significant role in facilitating the electron exchange between the electroactive center of GOx and the electrode. The GOx/Au/MXene/Nafion/GCE biosensor electrode displayed a linear amperometric response in the glucose concentration range from 0.1 to 18 mM with a relatively high sensitivity of 4.2 μAmM−1 cm−2 and a detection limit of 5.9 μM (S/N = 3). Furthermore, the biosensor exhibited excellent stability, reproducibility and repeatability. Therefore, the Au/MXene nanocomposite reported in this work is a potential candidate as an electrochemical transducer in electrochemical biosensors.

  12. Label-free electrochemical immunosensor based on cerium oxide nanowires for Vibrio cholerae O1 detection

    Energy Technology Data Exchange (ETDEWEB)

    Tam, Phuong Dinh, E-mail: phuongdinhtam@gmail.com; Thang, Cao Xuan, E-mail: thang.caoxuan@hust.edu.vn

    2016-01-01

    This paper developed a label-free immunosensor based on cerium oxide nanowire for Vibrio cholerae O1 detection application. The CeO{sub 2} nanowires were synthesized by hydrothermal reaction. The immobilization of Anti-V. cholerae O1 onto CeO{sub 2} nanowire-deposited sensor was performed via an amino ester, which was created by using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, and sulfo-N-hydroxysuccinimide. The electrochemical responses of the immunosensor were studied by electrochemical impedance spectroscopy with [Fe (CN) {sub 6}] {sup 3−/4−} as redox probe. A linear response in electron transfer resistance for cell of V. cholerae O1 concentration was found in the range of 1.0 × 10{sup 2} CFU/mL to 1.0 × 10{sup 4} CFU/mL. The detection limit of the immunosensor was 1.0 × 10{sup 2} CFU/mL. The immunosensor sensitivity was 56.82 Ω/CFU·mL{sup −1}. Furthermore, the parameters affecting immunosensor response were also investigated, as follows: pH value, immunoreaction time, incubation temperature, and anti-V. cholerae O1 concentration. - Highlights: • A label-free immunosensor based on cerium oxide nanowire for Vibrio cholerae O1 detection application was developed. • A linear response was found in the range of 1.0 × 10{sup 2} CFU/mL to 1.0 × 10{sup 4} CFU/mL. • The detection limit of the immunosensor was 1.0 × 10{sup 2} CFU/mL. • The immunosensor sensitivity was 56.82 Ω/CFU.mL{sup −1}.

  13. Single-walled carbon nanotube based transparent immunosensor for detection of a prostate cancer biomarker osteopontin

    Energy Technology Data Exchange (ETDEWEB)

    Sharma, Abhinav; Hong, Seongkyeol; Singh, Renu [School of Mechanical and Nuclear Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan 689-798 (Korea, Republic of); Jang, Jaesung, E-mail: jjang@unist.ac.kr [School of Mechanical and Nuclear Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan 689-798 (Korea, Republic of); Department of Biomedical Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan 689-798 (Korea, Republic of); School of Materials Science and Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan 689-798 (Korea, Republic of)

    2015-04-15

    Highlights: • A transparent CNT immunosensor is presented for detection of a prostate cancer biomarker osteopontin. • This immunosensor showed a highly linear and reproducible behavior from 1 pg mL{sup −1} to 1 μg mL{sup −1}. • The limit of detection of the immunosensor was 0.3 pg mL{sup −1}. • This immunosensor demonstrated high selectivity against bovine serum albumin and human serum. - Abstract: Osteopontin (OPN) is involved in almost all steps of cancer development, and it is being investigated as a potential biomarker for a diagnosis and prognosis of prostate cancer. Here, we report a label-free, highly sensitive and transparent immunosensor based on single-walled carbon nanotubes (SWCNTs) for detection of OPN. A high density of −COOH functionalized SWCNTs was deposited between two gold/indium tin oxide electrodes on a glass substrate by dielectrophoresis. Monoclonal antibodies specific to OPN were covalently immobilized on the SWCNTs. Relative resistance change of the immunosensors was measured as the concentration of OPN in phosphate buffer saline (PBS) and human serum was varied from 1 pg mL{sup −1} to 1 μg mL{sup −1} for different channel lengths of 2, 5, and 10 μm, showing a highly linear and reproducible behavior (R{sup 2} > 97%). These immunosensors were also specific to OPN against another test protein, bovine serum albumin, PBS and human serum, showing that a limit of detection for OPN was 0.3 pg mL{sup −1}. This highly sensitive and transparent immunosensor has a great potential as a simple point-of-care test kit for various protein biomarkers.

  14. Graphene-based immunosensor for electrochemical quantification of phosphorylated p53 (S15)

    Energy Technology Data Exchange (ETDEWEB)

    Xie Yunying; Chen Aiqiong [Key Laboratory of Pesticide and Chemical Biology of Ministry of Education, College of Chemistry, Central China Normal University, Wuhan 430079 (China); Du Dan, E-mail: dudan@mail.ccnu.edu.cn [Key Laboratory of Pesticide and Chemical Biology of Ministry of Education, College of Chemistry, Central China Normal University, Wuhan 430079 (China); Lin Yuehe, E-mail: Yuehe.lin@pnnl.gov [Pacific Northwest National Laboratory, Richland, WA 99352 (United States)

    2011-08-05

    Graphical abstract: A new graphene-based immunosensor for electrochemical quantification of phosphorylated p53 on serine 15 (phospho-p53{sup 15}) was proposed. This easily fabricated immunosensor provides a potential tool for analysis of other biomarkers. Highlights: {yields} A new graphene-based immunosensor for electrochemical quantification of phosphorylated p53 on serine 15 (phospho-p53{sup 15}) was proposed. {yields} Graphene served as sensor platform greatly increased detection sensitivity. {yields} This easily fabricated immunosensor provides a potential tool for analysis of other biomarkers. - Abstract: We reported a graphene-based immunosensor for electrochemical quantification of phosphorylated p53 on serine 15 (phospho-p53{sup 15}), a potential biomarker of gamma-radiation exposure. The principle is based on sandwich immunoassay and the resulting immunocomplex is formed among phospho-p53 capture antibody, phospho-p53{sup 15} antigen, biotinylated phospho-p53{sup 15} detection antibody and horseradish peroxidase (HRP)-labeled streptavidin. The introduced HRP results in an electrocatalytic response to reduction of hydrogen peroxide in the presence of thionine. Graphene served as sensor platform not only promotes electron transfer, but also increases the surface area to introduce a large amount of capture antibody, thus increasing the detection sensitivity. The experimental conditions including blocking agent, immunoreaction time and substrate concentration have been optimized. Under the optimum conditions, the increase of response current is proportional to the phospho-p53{sup 15} concentration in the range of 0.2-10 ng mL{sup -1}, with the detection limit of 0.1 ng mL{sup -1}. The developed immunosensor exhibits acceptable stability and reproducibility and the assay results for phospho-p53{sup 15} are in good correlation with the known values. This easily fabricated immunosensor provides a new promising tool for analysis of phospho-p53{sup 15} and other

  15. Measurement system for nitrous oxide based on amperometric gas sensor

    Science.gov (United States)

    Siswoyo, S.; Persaud, K. C.; Phillips, V. R.; Sneath, R.

    2017-03-01

    It has been well known that nitrous oxide is an important greenhouse gas, so monitoring and control of its concentration and emission is very important. In this work a nitrous oxide measurement system has been developed consisting of an amperometric sensor and an appropriate lab-made potentiostat that capable measuring picoampere current ranges. The sensor was constructed using a gold microelectrode as working electrode surrounded by a silver wire as quasi reference electrode, with tetraethyl ammonium perchlorate and dimethylsulphoxide as supporting electrolyte and solvent respectively. The lab-made potentiostat was built incorporating a transimpedance amplifier capable of picoampere measurements. This also incorporated a microcontroller based data acquisition system, controlled by a host personal computer using a dedicated computer program. The system was capable of detecting N2O concentrations down to 0.07 % v/v.

  16. Immunosensor Based on Surface Plasmon Resonance for Antigen Recognition

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    A novel immunosensor based on surface plasmon resonance(SPR)has been developed for the recognition of antigen.The sensor was designed on the basis of the fixed angle of incidence and measuring the reflected intensities in a wavelength range of 430-750 nm in real-time. An ultra-bright white light-emitting diode(LED)was used as the light source. Molecular self-assembling in solution was used to form the sensing membrane on gold substrate. It has been seen that the sensitivity of the SPR sensor with 3-mercaptopropionic acid(MPA)/protein A(SPA) sensing membrane is considerably higher than that with MPA or SPA modified Sensing membrane. The kinetic processes on the sensing membrane were studied. The human B factor(Bf), an activator of complement 3(C3), was recognized among the other antigens. This sensor can also be used for other antigen/antibody or adaptor/receptor recognition. Under optimized experimental conditions, the sensor has good selectivity, repeatability, and reversibility.

  17. Label-free bead-based metallothionein electrochemical immunosensor.

    Science.gov (United States)

    Nejdl, Lukas; Nguyen, Hoai Viet; Richtera, Lukas; Krizkova, Sona; Guran, Roman; Masarik, Michal; Hynek, David; Heger, Zbynek; Lundberg, Karin; Erikson, Kristofer; Adam, Vojtech; Kizek, Rene

    2015-08-01

    A novel microfluidic label-free bead-based metallothionein immunosensors was designed. To the surface of superparamagnetic agarose beads coated with protein A, polyclonal chicken IgY specifically recognizing metallothionein (MT) were immobilized via rabbit IgG. The Brdicka reaction was used for metallothionein detection in a microfluidic printed 3D chip. The assembled chip consisted of a single copper wire coated with a thin layer of amalgam as working electrode. Optimization of MT detection using designed microfluidic chip was performed in stationary system as well as in the flow arrangement at various flow rates (0-1800 μL/min). In stationary arrangement it is possible to detect MT concentrations up to 30 ng/mL level, flow arrangement allows reliable detection of even lower concentration (12.5 ng/mL). The assembled miniature flow chip was subsequently tested for the detection of MT elevated levels (at approx. level 100 μg/mL) in samples of patients with cancer. The stability of constructed device for metallothionein detection in flow arrangement was found to be several days without any maintenance needed.

  18. Electrochemical Immunosensor Based on Polythionine/Gold Nanoparticles for the Determination of Aflatoxin B1

    Directory of Open Access Journals (Sweden)

    Joseph H.O. Owino

    2008-12-01

    Full Text Available An aflatoxin B1 (AFB1 electrochemical immunosensor was developed by the immobilisation of aflatoxin B1-bovine serum albumin (AFB1-BSA conjugate on a polythionine (PTH/gold nanoparticles (AuNP-modified glassy carbon electrode (GCE. The surface of the AFB1-BSA conjugate was covered with horseradish peroxidase (HRP, in order to prevent non-specific binding of the immunosensors with ions in the test solution. The AFB1 immunosensor exhibited a quasi-reversible electrochemistry as indicated by a cyclic voltammetric (CV peak separation (ΔEp value of 62 mV. The experimental procedure for the detection of AFB1 involved the setting up of a competition between free AFB1 and the immobilised AFB1-BSA conjugate for the binding sites of free anti-aflatoxin B1 (anti-AFB1 antibody. The immunosensor’s differential pulse voltammetry (DPV responses (peak currents decreased as the concentration of free AFB1 increased within a dynamic linear range (DLR of 0.6 - 2.4 ng/mL AFB1 and a limit of detection (LOD of 0.07 ng/mL AFB1. This immunosensing procedure eliminates the need for enzyme-labeled secondary antibodies normally used in conventional ELISA–based immunosensors.

  19. Flow-injection amperometric glucose biosensors based on graphene/Nafion hybrid electrodes

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Bong Gill, E-mail: k1811@kaist.ac.kr [Department of Chemical and Biomolecular Engineering (BK21 Program), KAIST, Daejeon 305-701 (Korea, Republic of); Im, Jinkyu, E-mail: JINQ@paran.com [Department of Chemistry and Research Institute of Basic Sciences, Kyung Hee University, 1 Hoegidong, Dongdamoongu, Seoul (Korea, Republic of); Kim, Hoon Sik, E-mail: khs2004@khu.ac.kr [Department of Chemistry and Research Institute of Basic Sciences, Kyung Hee University, 1 Hoegidong, Dongdamoongu, Seoul (Korea, Republic of); Park, HoSeok, E-mail: phs0727@khu.ac.kr [Department of Chemical Engineering, College of Engineering, Kyung Hee University, 1 Seochon-dong, Giheung-gu, Youngin-si, Gyeonggi-do 446-701 (Korea, Republic of)

    2011-11-30

    Graphical abstract: Amperometric biosensors based on graphene hybrids showed the fast, sensitive, and stable amperometric responses in the flow injection system for automatically monitoring glucose. Display Omitted Highlights: > Flow-injection amperometric glucose biosensors were fabricated using reduced graphene oxide/Nafion hybrids. > The electrochemical kinetics of biosensors were comprehensively investigated by analysing electron transfer rate, charge transfer resistance, and ion diffusion coefficient, respectively. > The biosensors exhibited the fast, sensitive, and stable amperometric responses in the flow injection system for detecting glucose. - Abstract: In this research, we demonstrated the fabrication of flow-injection amperometric glucose biosensors based on RGO/Nafion hybrids. The nanohybridization of the reduced graphene oxide (RGO) by Nafion provided the fast electron transfer (ET) for the sensitive amperometric biosensor platforms. The ET rate (k{sub s}) and the charge transfer resistance (R{sub CT}) of GOx-RGO/Nafion hybrids were evaluated to verify the accelerated ET. Moreover, hybrid biosensors revealed a quasi-reversible and surface controlled process, as confirmed by the low peak-to-peak ({Delta}E{sub p}) and linear relations between I{sub p} and scan rate ({nu}). Hybrid biosensors showed the fast response time of {approx}3 s, the sensitivity of 3.8 {mu}A mM{sup -1} cm{sup -2}, the limit of detection of 170 {mu}M, and the linear detection range of 2-20 mM for the flow-injection amperometric detection of glucose. Furthermore, interference effect of oxidizable species such as ascorbic acid (AA) and uric acid (UA) on the performance of hybrid biosensors was prevented at the operating potential of -0.20 V even under the flow injection mode. Therefore, the fast, sensitive, and stable amperometric responses of hybrid biosensors in the flow injection system make it highly suitable for automatically monitoring glucose.

  20. Investigate electrochemical immunosensor of cortisol based on gold nanoparticles/magnetic functionalized reduced graphene oxide.

    Science.gov (United States)

    Sun, Bolu; Gou, Yuqiang; Ma, Yuling; Zheng, Xiaoping; Bai, Ruibin; Ahmed Abdelmoaty, Ahmed Attia; Hu, Fangdi

    2017-02-15

    A sensitively competitive electrochemical immunosensor for the detection of cortisol was successfully developed based on gold nanoparticles and magnetic functionalized reduced graphene oxide (AuNPs/MrGO). In order to construct the base of the immunosensor, the MrGO was initially fabricated by chemical cross-linking and used to modify the nafion pretreated glassy carbon electrode. Subsequently, the surface of electrode was modified by AuNPs via electrochemical deposition. A variety of cortisol (Cor) can be firmly loaded in the AuNPs/MrGO with large specific surface area and good bioactivity to construct the basic electrode (Cor/AuNPs/MrGO/Nafion@GCE), which was characterized by the cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS), respectively. Due to the cortisol on the surface of basic electrode and samples can competitively combine with the cortisol antibody labelled by horseradish peroxidase (HRP-Strept-Biotin-Ab). Finally, the detection signal of electrochemical immunosensor (HRP-Strept-Biotin-Ab-Cor/AuNPs/MrGO/Nafion@GCE) in the test liquid had negative correlations with the concentration of cortisol in samples. The AuNPs/MrGO with excellent electrical conductivity being applied, the electrochemical response of the immunosensor was immensely amplified. The immunosensor displayed excellent analytical performance for the detection of cortisol range from 0.1 to 1000ng/mL with a detection limit of 0.05ng/mL at 3σ. Moreover, compared the developed immunoassay with commercially available enzyme linked immunosorbent assay, the proposed method showed good precision, acceptable stability and reproducibility, indicating the immunosensor could be used for the sensitive, efficient and real-time detection of cortisol in real samples. Therefore, the present strategy provides a novel and convenient method for clinical determination of cortisol.

  1. A novel electrochemical immunosensor based on magnetosomes for detection of staphylococcal enterotoxin B in milk.

    Science.gov (United States)

    Wu, Longyun; Gao, Bo; Zhang, Fang; Sun, Xiulan; Zhang, Yinzhi; Li, Zaijun

    2013-03-15

    In this paper, a novel electrochemical immunosensor to detect staphylococcal enterotoxin B based on bio-magnetosomes, polyaniline nano-gold composite and 1,2-dimethyl-3-butylimidazolium hexafluorophosphate ionic liquid, was developed, and found to exhibit high sensitivity and stability. The specific antibody to staphylococcal enterotoxin B conjugated with the magnetosomes showed rapid immunoreactions and good dispersion, which contributed to the formation of a nanostructurally smooth and dense film on the surface of a gold electrode. Polyaniline nano-gold composite and 1,2-dimethyl-3-butylimidazolium hexafluorophosphate ionic liquid were used to modify the electrode as mediators to improve the electron transfer and offer an excellent biocompatible microenvironment for the antibody to retain its activity to enhance the response of the electrochemical sensor. Under optimal conditions, the developed immunosensor showed a good linear response in the range from 0.05 to 5 ng/mL (R(2)=0.9957) with a detection limit as low as 0.017 ng/mL, compared with the one without magnetosomes (0.05-5 ng/mL, 0.033 ng/mL), this developed immunosensor showed a wider response range and a reduced detection limit. And a good specificity with little adsorption to staphylococcal enterotoxin A, C and Na(+), K(+), Ca(2+) was obtained. Moreover, the immunosensor exhibited a good long-time stability at 4 °C reaching up to 60 days, which showed a relatively long working life. Meanwhile the immunosensor could be regenerated four times using NaOH elution. The sensor also displayed a good repeatability with a relative standard deviation of 5.02% for staphylococcal enterotoxin B detection (1 ng/mL, n=9). Furthermore, high recoveries in milk samples from 81% to 118% were achieved and successfully applied to milk sample detection. The obtained results demonstrate that the developed electrochemical immunosensor is a promising tool for the detection of staphylococcal enterotoxin B in food.

  2. Microfabricated, amperometric, enzyme-based biosensors for in vivo applications.

    Science.gov (United States)

    Weltin, Andreas; Kieninger, Jochen; Urban, Gerald A

    2016-07-01

    Miniaturized electrochemical in vivo biosensors allow the measurement of fast extracellular dynamics of neurotransmitter and energy metabolism directly in the tissue. Enzyme-based amperometric biosensing is characterized by high specificity and precision as well as high spatial and temporal resolution. Aside from glucose monitoring, many systems have been introduced mainly for application in the central nervous system in animal models. We compare the microsensor principle with other methods applied in biomedical research to show advantages and drawbacks. Electrochemical sensor systems are easily miniaturized and fabricated by microtechnology processes. We review different microfabrication approaches for in vivo sensor platforms, ranging from simple modified wires and fibres to fully microfabricated systems on silicon, ceramic or polymer substrates. The various immobilization methods for the enzyme such as chemical cross-linking and entrapment in polymer membranes are discussed. The resulting sensor performance is compared in detail. We also examine different concepts to reject interfering substances by additional membranes, aspects of instrumentation and biocompatibility. Practical considerations are elaborated, and conclusions for future developments are presented. Graphical Abstract ᅟ.

  3. Photonic crystal fiber-based immunosensor for high-performance detection of alpha fetoprotein.

    Science.gov (United States)

    Liu, Xiaoxia; Song, Xingda; Dong, Zhiyong; Meng, Xiaoting; Chen, Yiping; Yang, Li

    2017-05-15

    We have developed a sensitive photonic crystal fiber (PCF)-based immunosensor for detection of alpha fetoprotein (AFP). The unique PCF possesses a morphology characterized by numerous pore structures and a large surface area-to-volume ratio, which can be used as an immune-reaction carrier to improve the sensitivity and reaction speed of AFP detection. The PCF-based immunosensor possesses a low limit of detection of 0.1ng/mL, which is five times lower than that of the capillary-based sensor and 35 times lower than that of the traditional enzyme-linked immunosorbent assay. The wide linear dynamic range of 0.1-150ng/mL makes the developed immunosensor suitable for clinical practice. The proposed method was successfully applied to AFP detection in a clinical serum sample with acceptable precision. It is indicated that the present PCF-based immunosensor could be used as an attractive analytical platform for sensitive and specific detection of cancer biomarkers.

  4. Design and Development of a Microfluidic Amperometric Immunosensor for the Quantitative Detection of 2,6-dichlorobenzamide (BAM) Herbicide Residue in Ground Water

    DEFF Research Database (Denmark)

    Uthuppu, Basil

    and it is a novel characteristic of the microfluidic device prototype. The microfluidic device was automated using Lego® Mindstorms® servomotors to control its micro pumps and valves. By confirming (amperometrically) the regeneration capability of the optimized immunosurface and generating a standard curve for BAM...... the optimized BAM immunoassay and the electrochemical detection method. A modular approach was adopted for the fabrication of the microfluidic platform in order to make the device simple to integrate, automate and maintain. The microfluidic platform has an in-built micro flow-injection analysis (µFIA) system...

  5. A review of enzymatic uric acid biosensors based on amperometric detection.

    Science.gov (United States)

    Erden, Pınar Esra; Kılıç, Esma

    2013-03-30

    This review summarizes the studies carried on the development of amperometric uric acid biosensors over the past twenty years. Sensing principles, enzyme immobilization techniques, the electrode types, different approaches and various matrices used for biosensor fabrication are presented along with their benefits and limitations. Uric acid biosensors based on different modes of transducing devices such as optical, potentiometric, conductometric are also referred.

  6. An electrochemical immunosensor based on interdigitated array microelectrode for the detection of chlorpyrifos.

    Science.gov (United States)

    Cao, Yaoyao; Sun, Xia; Guo, Yemin; Zhao, Wenping; Wang, Xiangyou

    2015-02-01

    An electrochemical immunosensor based on interdigitated array microelectrodes (IDAMs) was developed for sensitive, specific and rapid detection of chlorpyrifos. Anti-chlorpyrifos monoclonal antibodies were orientedly immobilized onto the gold microelectrode surface through protein A. Chlorpyrifos were then captured by the immobilized antibody, resulting in an impedance change in the IDAMs surface. Electrochemical impedance spectroscopy was used in conjunction with the fabricated sensor to detect chlorpyrifos. Under optimum conditions, the impedance value change of chlorpyrifos was proportional to its concentrations in the range of 10(0)-10(5) ng/mL. The detection limit was found to be 0.014 ng/mL for chlorpyrifos. The proposed chlorpyrifos immunosensor could be used as a screening method in pesticide determination for the analysis of environmental, agricultural and pharmaceutical samples due to its rapidity, sensitivity and low cost.

  7. Graphene-based immunosensor for electrochemical quantification of phosphorylated p53 (S15)

    Energy Technology Data Exchange (ETDEWEB)

    Xie, Yunying; Chen, Aiqiong; Du, Dan; Lin, Yuehe

    2011-08-01

    We reported a graphene-based immunosensor for electrochemical quantification of phosphorylated p53 on serine 15 (phospho-p5315), a potential biomarker of gamma-radiation exposure. The principle is based on sandwich immunoassay and the resulting immunocomplex is formed among phospho-p53 capture antibody, phospho-p5315 antigen, biotinylated phospho-p5315 detection antibody and horseradish peroxidase (HRP)-labeled streptavidin. The introduced HRP results in an electrocatalytic response to reduction of hydrogen peroxide in the presence of thionine. Graphene served as sensor platform not only promotes electron transfer, but also increases the surface area to introduce a large amount of capture antibody, thus increasing the detection sensitivity. The experimental conditions including blocking agent, immunoreaction time and substrate concentration have been optimized. Under the optimum conditions, the increase of response current is proportional to the phospho-p5315 concentration in the range of 0.2–10 ng mL-1, with the detection limit of 0.1 ng mL-1. The developed immunosensor exhibits acceptable stability and reproducibility and the assay results for phospho-p5315 are in good correlation with the known values. This easily fabricated immunosensor provides a new promising tool for analysis of phospho-p5315 and other phosphorylated proteins.

  8. A Fumonisins Immunosensor Based on Polyanilino-Carbon Nanotubes Doped with Palladium Telluride Quantum Dots

    Directory of Open Access Journals (Sweden)

    Milua Masikini

    2014-12-01

    Full Text Available An impedimetric immunosensor for fumonisins was developed based on poly(2,5-dimethoxyaniline-multi-wall carbon nanotubes doped with palladium telluride quantum dots onto a glassy carbon surface. The composite was assembled by a layer-by-layer method to form a multilayer film of quantum dots (QDs and poly(2,5-dimethoxyaniline-multi-wall carbon nanotubes (PDMA-MWCNT. Preparation of the electrochemical immunosensor for fumonisins involved drop-coating of fumonisins antibody onto the composite modified glassy carbon electrode. The electrochemical impedance spectroscopy response of the FB1 immunosensor (GCE/PT-PDMA-MWCNT/anti-Fms-BSA gave a linear range of 7 to 49 ng L−1 and the corresponding sensitivity and detection limits were 0.0162 kΩ L ng−1 and 0.46 pg L−1, respectively, hence the limit of detection of the GCE/PT-PDMA-MWCNT immunosensor for fumonisins in corn certified material was calculated to be 0.014 and 0.011 ppm for FB1, and FB2 and FB3, respectively. These results are lower than those obtained by ELISA, a provisional maximum tolerable daily intake (PMTDI for fumonisins (the sum of FB1, FB2, and FB3 established by the Joint FAO/WHO expert committee on food additives and contaminants of 2 μg kg−1 and the maximum level recommended by the U.S. Food and Drug Administration (FDA for protection of human consumption (2–4 mg L−1.

  9. Side-polished fiber immunosensor based on surface plasmon resonance for detection of Legionella pneumophila

    Science.gov (United States)

    Tsao, Yu-Chia; Yang, Yi-Wen; Tsai, Woo-Hu; Yan, Tsong-Rong

    2008-02-01

    Side-polished fiber immunosensor based on surface plasmon resonance (SPR) onto self-assembled protein A layer was proposed for the detection of Legionella pneumophila. A self-assembled protein A layer on gold (Au) surface was fabricated by adsorbing a mixture of 11-mercaptoundecanoic acid (MUA) and activated by N-Ethyl-N'-(3-dimethylaminopropyl) carbodiimide/ N-Hydroxysuccinimide (EDC/NHS). The formation of self-assembled protein A and gold layer on side-polished surface and the binding of antibody and antigen in series were confirmed by SPR response on spectrum. The binding protein A layer can improve the sensitivity, which indirectly supports the configurations that antibody layer is immobilized on the binding protein A layer with a well-ordered orientation. The surface morphology analyses of self-assembled protein A layer on Au substrate and monoclonal antibody against L. pneumophila immobilized on protein A were demonstrated by SPR dip shifts on optical spectrum analyzer. The SPR fiber immunosensor for detection of L. pneumophila was developed and the detection limit was 10 CFU/ml with the SPR dip shift in wavelength from 1070 to 1105nm. The current fabrication technique of a SPR immunosensor using optical fiber for the detection of Legionella pneumophila could be applied to construct other biosensor.

  10. Two-dimensional paper chromatography-based fluorescent immunosensor for detecting acute myocardial infarction markers.

    Science.gov (United States)

    Cho, Jung-Hwan; Kim, Min-Ha; Mok, Rak-Sun; Jeon, Jin-Woo; Lim, Guei-Sam; Chai, Chan-Young; Paek, Se-Hwan

    2014-09-15

    A novel washing scheme following antigen-antibody reactions with analyte was used during construction of a fluorescent immunosensor to resolve the background problem in the lateral flow assay with human serum. An immuno-membrane strip was devised to simultaneously measure cardiac troponin I (cTnI), creatinine kinase-MB isoform (CK-MB), and myoglobin to diagnose acute myocardial infarction. This strip was then installed within a cartridge containing a built-in washing solution tank, which was used to supply the solution across the signal generation pad of the strip after the immune reactions. Such cross-flow washing was initiated by onset-signaling from the internal control and began to run automatically upon sample addition. Under optimal conditions, the immunosensor displayed a stably suppressed background baseline, enabling us to attain a low detection limit for cTnI (0.05 ng/mL) as well as favorable reproducibility for repetitive measurements (relative standard deviation 0.98. This result suggests that the new immunosensor system based on two-dimensional chromatography can be used for clinical testing.

  11. Quantum dots based electrochemiluminescent immunosensor by coupling enzymatic amplification for ultrasensitive detection of clenbuterol.

    Science.gov (United States)

    Yao, Xun; Yan, Panpan; Tang, Qinghui; Deng, Anping; Li, Jianguo

    2013-10-10

    An ultrasensitive electrochemiluminescence (ECL) immunosensor based on CdSe quantum dots (QDs) has been designed for the detection of clenbuterol. The immunosensor was fabricated by layer by layer and characterized with atomic force microscopic images (AFM) and electrochemical impedance spectra (EIS). In oxygen-saturated pH=9.0 Tris-HCl buffer, a strong ECL emission of QDs could be observed during the cathodic process due to the H2O2 product from electrochemical reduction of dissolved oxygen. Upon the formation of immunocomplex, the second antibody labeled with horseradish peroxidase was simply immobilized on the electrode surface. The ECL emission decreased since steric hindrance of the immunocomplex slowed down the electron-transfer speed of dissolved oxygen, and also could be greatly amplified by an enzymatic cycle to consume the self-produced coreactant. Using clenbuterol as model analyte, the ECL intensity was determined by the concentration of competitive immunoassay of clenbuterol with a wide calibration in the range of 0.05 ng mL(-1) to 1000 ng mL(-1), and a low detection limit was 0.02 ng mL(-1). The immunosensor shows good stability and fabrication reproducibility. It was applied to detecting practical samples with the satisfactory results. This immunosensing strategy opens a new avenue for detection of residue and application of QDs in ECL biosensing.

  12. Immunosensor based on carbon nanotube/manganese dioxide electrochemical tags.

    Science.gov (United States)

    Tu, Meng-Che; Chen, Han-Yi; Wang, Yuxi; Moochhala, Shabbir M; Alagappan, Palaniappan; Liedberg, Bo

    2015-01-01

    This article reports on carbon nanotube/manganese dioxide (CNT-MnO2) composites as electrochemical tags for non-enzymatic signal amplification in immunosensing. The synthesized CNT-MnO2 composites showed good electrochemical activity, electrical conductivity and stability. The electrochemical signal of CNT-MnO2 composites coated glassy carbon electrode (GCE) increased by nearly two orders of magnitude compared to bare GCE in hydrogen peroxide (H2O2) environment. CNT-MnO2 composite was subsequently validated as electrochemical tags for sensitive detection of α-fetoprotein (AFP), a tumor marker for diagnosing hepatocellular carcinoma. The electrochemical immunosensor demonstrated a linear response on a log-scale for AFP concentrations ranging from 0.2 to 100 ng mL(-1). The limit of detection (LOD) was estimated to be 40 pg mL(-1) (S/N=3) in PBS buffer. Further measurements using AFP spiked plasma samples revealed the applicability of fabricated CNT-MnO2 composites for clinical and diagnostic applications.

  13. [Determination of a Candida albicans antigen using an amperometric immunoenzyme sensor].

    Science.gov (United States)

    Kutyreva, M P; Mediantseva, E P; Khaldeeva, E V; Glushko, N I; Budnikov, G K

    1998-01-01

    Determination new variant enzyme immunoassay with amperometric enzyme immunosensor, including the immobilizing enzyme-choline esterase and antibodies against Candida albicans (CA) in biosensitivity part of sensor, for diagnose disease of CA. The method for determination of CA based on combination immunochemical reactions and voltammetric indication of analytical signal was developed. Amperometric enzyme immunosensor developed has been used as detector. Differences dilutions of antibody (Ab) against antigen (Ag) of CA immobilizing in common with choline esterase (CE). The method of immobilization developed allows to receive the sensor with including the immobilized CE and Ab in common. The method of determination of CA based on combination the reaction of forming immune complex tAb-AgI with enzyme immunosensor for its detection. The dynamic range of concentrations determined of Ag depends on degree of dilution of Ab used for manufactory biosensitivity part of sensor. The data indicate that the [Ab-Ag] immune complexes are stable. This is also confirmed by the values of [Ab-Ag] binding constants, obtained in Scatchard coordinates. This method of determination doesn't require special preparation of a sample. Selectivity, sensitivity, simplicity and quickness are characterize of this method which could be used for manufacturing test-sistem for determination CA in blood.

  14. A Label-Free Electrochemical Immunosensor for Carbofuran Detection Based on a Sol-Gel Entrapped Antibody

    Directory of Open Access Journals (Sweden)

    Qingqing Li

    2011-10-01

    Full Text Available In this study, an anti-carbofuran monoclonal antibody (Ab was immobilized on the surface of a glassy carbon electrode (GCE using silica sol-gel (SiSG technology. Thus, a sensitive, label-free electrochemical immunosensor for the direct determination of carbofuran was developed. The electrochemical performance of immunoreaction of antigen with the anti-carbofuran monoclonal antibody was investigated by cyclic voltammetry (CV and electrochemical impedance spectroscopy (EIS, in which phosphate buffer solution containing [Fe(CN6]3−/4− was used as the base solution for test. Because the complex formed by the immunoreaction hindered the diffusion of [Fe(CN6]3−/4− on the electrode surface, the redox peak current of the immunosensor in the CV obviously decreased with the increase of the carbofuran concentration. The pH of working solution, the concentration of Ab and the incubation time of carbofuran were studied to ensure the sensitivity and conductivity of the immunosensor. Under the optimal conditions, the linear range of the proposed immunosensor for the determination of carbofuran was from 1 ng/mL to 100 μg/mL and from 50 μg/mL to 200 μg/mL with a detection limit of 0.33 ng/mL (S/N = 3. The proposed immunosensor exhibited good high sensitivity and stability, and it was thus suitable for trace detection of carbofuran pesticide residues.

  15. A label-free electrochemical immunosensor for carbofuran detection based on a sol-gel entrapped antibody.

    Science.gov (United States)

    Sun, Xia; Du, Shuyuan; Wang, Xiangyou; Zhao, Wenping; Li, Qingqing

    2011-01-01

    In this study, an anti-carbofuran monoclonal antibody (Ab) was immobilized on the surface of a glassy carbon electrode (GCE) using silica sol-gel (SiSG) technology. Thus, a sensitive, label-free electrochemical immunosensor for the direct determination of carbofuran was developed. The electrochemical performance of immunoreaction of antigen with the anti-carbofuran monoclonal antibody was investigated by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS), in which phosphate buffer solution containing [Fe(CN)(6)](3-/4-) was used as the base solution for test. Because the complex formed by the immunoreaction hindered the diffusion of [Fe(CN)(6)](3-/4-) on the electrode surface, the redox peak current of the immunosensor in the CV obviously decreased with the increase of the carbofuran concentration. The pH of working solution, the concentration of Ab and the incubation time of carbofuran were studied to ensure the sensitivity and conductivity of the immunosensor. Under the optimal conditions, the linear range of the proposed immunosensor for the determination of carbofuran was from 1 ng/mL to 100 μg/mL and from 50 μg/mL to 200 μg/mL with a detection limit of 0.33 ng/mL (S/N = 3). The proposed immunosensor exhibited good high sensitivity and stability, and it was thus suitable for trace detection of carbofuran pesticide residues.

  16. Solid-state voltammetry-based electrochemical immunosensor for Escherichia coli using graphene oxide-Ag nanoparticle composites as labels.

    Science.gov (United States)

    Jiang, Xiaochun; Chen, Kun; Wang, Jing; Shao, Kang; Fu, Tao; Shao, Feng; Lu, Donglian; Liang, Jiangong; Foda, M Frahat; Han, Heyou

    2013-06-21

    A new electrochemical immunosensor based on solid-state voltammetry was fabricated for the detection of Escherichia coli (E. coli) by using graphene oxide-Ag nanoparticle composites (P-GO-Ag) as labels. To construct the platform, Au nanoparticles (AuNPs) were first self-assembled on an Au electrode surface through cysteamine and served as an effective matrix for antibody (Ab) attachment. Under a sandwich-type immunoassay format, the analyte and the probe (P-GO-Ag-Ab) were successively captured onto the immunosensor. Finally, the bonded AgNPs were detected through a solid-state redox process in 0.2 M of KCl solution. Combining the advantages of the high-loading capability of graphene oxide with promoted electron-transfer rate of AuNPs, this immunosensor produced a 26.92-fold signal enhancement compared with the unamplified protocol. Under the optimal conditions, the immunosensor exhibited a wide linear dependence on the logarithm of the concentration of E. coli ranging from 50 to 1.0 × 10(6) cfu mL(-1) with a detection limit of 10 cfu mL(-1). Moreover, as a practical application, the proposed immunosensor was used to monitor E. coli in lake water with satisfactory results.

  17. Immunosensor based on the ZnO nanorod networks for the detection of H1N1 swine influenza virus.

    Science.gov (United States)

    Jang, Yunseok; Park, Jungil; Pak, Youngmi Kim; Pak, James Jungho

    2012-07-01

    This paper presents an immunosensor fabricated on patterned zinc oxide nanorod networks (ZNNs) for detecting the H1N1 swine influenza virus (H1N1 SIV). Nanostructured ZnO with a high isoelectric point (IEP, approximately 9.5) possesses good absorbability for proteins with low IEPs. Hydrothermally grown ZNNs were fabricated on a patterned Au electrode (0.02 cm2) through a lift-off process. To detect the H1N1 SIV, the sandwich enzyme-linked immunosorbent assay (ELISA) method was employed in the immunosensor. The immunosensor was evaluated in an acetate buffer solution containing 3,3',5,5'-tetramethylbenzidine (TMB) via cyclic voltammetry at various H1N1 SIV concentrations (1 pg/mL-5 ng/mL). The measurement results of the fabricated immunosensor showed that the reduction currents of TMB at 0.25 V logarithmically increased from 259.37 to 577.98 nA as the H1N1 SIV concentration changed from 1 pg/mL to 5 ng/mL. An H1N1 SIV immunosensor, based on the patterned ZNNs, was successfully realized for detecting 1 pg/mL-5 ng/mL H1N1 SIV concentrations, with a detection limit of 1 pg/mL for H1N1 SIV.

  18. Automated microfluidic platform of bead-based electrochemical immunosensor integrated with bioreactor for continual monitoring of cell secreted biomarkers

    Science.gov (United States)

    Riahi, Reza; Shaegh, Seyed Ali Mousavi; Ghaderi, Masoumeh; Zhang, Yu Shrike; Shin, Su Ryon; Aleman, Julio; Massa, Solange; Kim, Duckjin; Dokmeci, Mehmet Remzi; Khademhosseini, Ali

    2016-04-01

    There is an increasing interest in developing microfluidic bioreactors and organs-on-a-chip platforms combined with sensing capabilities for continual monitoring of cell-secreted biomarkers. Conventional approaches such as ELISA and mass spectroscopy cannot satisfy the needs of continual monitoring as they are labor-intensive and not easily integrable with low-volume bioreactors. This paper reports on the development of an automated microfluidic bead-based electrochemical immunosensor for in-line measurement of cell-secreted biomarkers. For the operation of the multi-use immunosensor, disposable magnetic microbeads were used to immobilize biomarker-recognition molecules. Microvalves were further integrated in the microfluidic immunosensor chip to achieve programmable operations of the immunoassay including bead loading and unloading, binding, washing, and electrochemical sensing. The platform allowed convenient integration of the immunosensor with liver-on-chips to carry out continual quantification of biomarkers secreted from hepatocytes. Transferrin and albumin productions were monitored during a 5-day hepatotoxicity assessment in which human primary hepatocytes cultured in the bioreactor were treated with acetaminophen. Taken together, our unique microfluidic immunosensor provides a new platform for in-line detection of biomarkers in low volumes and long-term in vitro assessments of cellular functions in microfluidic bioreactors and organs-on-chips.

  19. Automated microfluidic platform of bead-based electrochemical immunosensor integrated with bioreactor for continual monitoring of cell secreted biomarkers.

    Science.gov (United States)

    Riahi, Reza; Shaegh, Seyed Ali Mousavi; Ghaderi, Masoumeh; Zhang, Yu Shrike; Shin, Su Ryon; Aleman, Julio; Massa, Solange; Kim, Duckjin; Dokmeci, Mehmet Remzi; Khademhosseini, Ali

    2016-04-21

    There is an increasing interest in developing microfluidic bioreactors and organs-on-a-chip platforms combined with sensing capabilities for continual monitoring of cell-secreted biomarkers. Conventional approaches such as ELISA and mass spectroscopy cannot satisfy the needs of continual monitoring as they are labor-intensive and not easily integrable with low-volume bioreactors. This paper reports on the development of an automated microfluidic bead-based electrochemical immunosensor for in-line measurement of cell-secreted biomarkers. For the operation of the multi-use immunosensor, disposable magnetic microbeads were used to immobilize biomarker-recognition molecules. Microvalves were further integrated in the microfluidic immunosensor chip to achieve programmable operations of the immunoassay including bead loading and unloading, binding, washing, and electrochemical sensing. The platform allowed convenient integration of the immunosensor with liver-on-chips to carry out continual quantification of biomarkers secreted from hepatocytes. Transferrin and albumin productions were monitored during a 5-day hepatotoxicity assessment in which human primary hepatocytes cultured in the bioreactor were treated with acetaminophen. Taken together, our unique microfluidic immunosensor provides a new platform for in-line detection of biomarkers in low volumes and long-term in vitro assessments of cellular functions in microfluidic bioreactors and organs-on-chips.

  20. Visible-light driven photoelectrochemical immunosensor for insulin detection based on MWCNTs@SnS2@CdS nanocomposites.

    Science.gov (United States)

    Liu, Yixin; Zhang, Yifeng; Wu, Dan; Fan, Dawei; Pang, Xuehui; Zhang, Yong; Ma, Hongmin; Sun, Xu; Wei, Qin

    2016-12-15

    In this work, a label-free photoelectrochemical (PEC) immunosensor was developed for ultrasensitive detection of insulin based on MWCNTs@SnS2@CdS nanocomposites. As graphene-like 2D nanomaterial, SnS2 nanosheets loaded on the conducting framework of multi-walled carbon nanotubes (MWCNTs) were adopted for the construction of immunosensor for the first time, providing a favorable substrate for in-situ growth of CdS nanocrystal that had suitable band structure matching well with SnS2. The well-matched band structure of these two metal sulfides effectively inhibited the recombination of photogenerated electron-hole pairs, thus improving the photo-to-current conversion efficiency. Besides, the introduction of MWCNTs facilitated electron transfer across the surface of electrodes, leading to a further increment of photocurrent. The as constructed label-free PEC immunosensor based on MWCNTs@SnS2@CdS nanocomposites exhibited excellent PEC performance for the detection of insulin. The concentrations of insulin could be directly detected based on the decrement of photocurrent that was brought by the increased steric hindrances due to the formation of antigen-antibody immunocomplexes. Under the optimal conditions, the PEC immunosensor had a sensitive response to insulin in a linear range of 0.1pgmL(-1) to 5ngmL(-1) with a detection limit of 0.03pgmL(-1). Meanwhile, good stability and selectivity were achieved as well. The design and fabrication of this PEC immunosensor based on MWCNTs@SnS2@CdS nanocomposites not only provided an ideal platform for the detection of insulin, but also opened up a new avenue for the development of immunosensor for some other biomarkers analysis.

  1. A Nonenzymatic Electrochemical Immunosensor for Ultrasensitive Detection of Tumor Biomarkers Based on Palladium Nanoparticles Conjugated Reduced Graphene Nanosheets.

    Science.gov (United States)

    Li, WenTing; Li, YiSong; Wu, YongKang; Liao, JinFeng; Qi, TingTing; Li, He; Li, Jun; Zhang, XiaoNing; Qian, ZhiYong

    2015-11-01

    A nonenzymatic electrochemical immunosensor based on palladium nanoparticles conjugated reduced graphene nanosheets (Pd-GS) for sensitive detection of cancer biomarker a-fetoprotein (AFP) is described. Primary antibody-AFP (Ab1) was immobilized onto the surface of reduced graphene nanosheets (rGO) through an amidation reaction between the carboxylic acid group of the rGO and the available amine groups of Ab1. Pd-GS which was prepared by one-spot synthesis is employed to immobilize secondary antibody (Ab2). The resulting Pd-GS-Ab2 conjugate was used as a label for the immunosensor to detect AFP. The amplified immunoassay exhibits high sensitivity, wide linear rang (0.01-10 ng/mL), low detection limit (3.0 pg/mL), acceptable stability and reproducibility. And such immunosensor also shows good recovery in the assay results for AFP in human serum samples.

  2. Critical evaluation of acetylthiocholine iodide and acetylthiocholine chloride as substrates for amperometric biosensors based on acetylcholinesterase.

    Science.gov (United States)

    Bucur, Madalina-Petruta; Bucur, Bogdan; Radu, Gabriel-Lucian

    2013-01-25

    Numerous amperometric biosensors have been developed for the fast analysis of neurotoxic insecticides based on inhibition of cholinesterase (AChE). The analytical signal is quantified by the oxidation of the thiocholine that is produced enzymatically by the hydrolysis of the acetylthiocholine pseudosubstrate. The pseudosubstrate is a cation and it is associated with chloride or iodide as corresponding anion to form a salt. The iodide salt is cheaper, but it is electrochemically active and consequently more difficult to use in electrochemical analytical devices. We investigate the possibility of using acetylthiocholine iodide as pseudosubstrate for amperometric detection. Our investigation demonstrates that operational conditions for any amperometric biosensor that use acetylthiocholine iodide must be thoroughly optimized to avoid false analytical signals or a reduced sensitivity. The working overpotential determined for different screen-printed electrodes was: carbon-nanotubes (360 mV), platinum (560 mV), gold (370 mV, based on a catalytic effect of iodide) or cobalt phthalocyanine (110 mV, but with a significant reduced sensitivity in the presence of iodide anions).

  3. Quantum dots based electrochemiluminescent immunosensor by coupling enzymatic amplification for ultrasensitive detection of clenbuterol

    Energy Technology Data Exchange (ETDEWEB)

    Yao, Xun; Yan, Panpan; Tang, Qinghui [College of Chemistry, Chemical Engineering and Materials Science, Soochow University, Suzhou 215123 (China); The Key Lab of Health Chemistry and Molecular Diagnosis of Suzhou, Suzhou 215123 (China); Deng, Anping, E-mail: denganping@suda.edu.cn [College of Chemistry, Chemical Engineering and Materials Science, Soochow University, Suzhou 215123 (China); The Key Lab of Health Chemistry and Molecular Diagnosis of Suzhou, Suzhou 215123 (China); Li, Jianguo, E-mail: lijgsd@suda.edu.cn [College of Chemistry, Chemical Engineering and Materials Science, Soochow University, Suzhou 215123 (China); The Key Lab of Health Chemistry and Molecular Diagnosis of Suzhou, Suzhou 215123 (China)

    2013-10-10

    Graphical abstract: -- Highlights: •An ultrasensitive ECL immunosensor of CdSe QDs for clenbuterol determination is developed. •The CdSe QDs showed great biocompatibility and could be easier to make direct use of such QDs in the region of biological system. •Enzymatic amplification strategy was proposed by combining the coreactant and pAb/GaRIgG-HRP. •Enzymatic amplification increased ECL emission and extended the analyte in presence of substrate. •It provided a method for detecting clenbuterol and enlarged the usage of QDs in ECL biosensing. -- Abstract: An ultrasensitive electrochemiluminescence (ECL) immunosensor based on CdSe quantum dots (QDs) has been designed for the detection of clenbuterol. The immunosensor was fabricated by layer by layer and characterized with atomic force microscopic images (AFM) and electrochemical impedance spectra (EIS). In oxygen-saturated pH = 9.0 Tris-HCl buffer, a strong ECL emission of QDs could be observed during the cathodic process due to the H{sub 2}O{sub 2} product from electrochemical reduction of dissolved oxygen. Upon the formation of immunocomplex, the second antibody labeled with horseradish peroxidase was simply immobilized on the electrode surface. The ECL emission decreased since steric hindrance of the immunocomplex slowed down the electron-transfer speed of dissolved oxygen, and also could be greatly amplified by an enzymatic cycle to consume the self-produced coreactant. Using clenbuterol as model analyte, the ECL intensity was determined by the concentration of competitive immunoassay of clenbuterol with a wide calibration in the range of 0.05 ng mL{sup −1} to 1000 ng mL{sup −1}, and a low detection limit was 0.02 ng mL{sup −1}. The immunosensor shows good stability and fabrication reproducibility. It was applied to detecting practical samples with the satisfactory results. This immunosensing strategy opens a new avenue for detection of residue and application of QDs in ECL biosensing.

  4. Electrochemical signal amplification for immunosensor based on 3D interdigitated array electrodes.

    Science.gov (United States)

    Han, Donghoon; Kim, Yang-Rae; Kang, Chung Mu; Chung, Taek Dong

    2014-06-17

    We devised an electrochemical redox cycling based on three-dimensional interdigitated array (3D IDA) electrodes for signal amplification to enhance the sensitivity of chip-based immunosensors. The 3D IDA consists of two closely spaced parallel indium tin oxide (ITO) electrodes that are positioned not only on the bottom but also the ceiling, facing each other along a microfluidic channel. We investigated the signal intensities from various geometric configurations: Open-2D IDA, Closed-2D IDA, and 3D IDA through electrochemical experiments and finite-element simulations. The 3D IDA among the four different systems exhibited the greatest signal amplification resulting from efficient redox cycling of electroactive species confined in the microchannel so that the faradaic current was augmented by a factor of ∼100. We exploited the enhanced sensitivity of the 3D IDA to build up a chronocoulometric immunosensing platform based on the sandwich enzyme-linked immunosorbent assay (ELISA) protocol. The mouse IgGs on the 3D IDA showed much lower detection limits than on the Closed-2D IDA. The detection limit for mouse IgG measured using the 3D IDA was ∼10 fg/mL, while it was ∼100 fg/mL for the Closed-2D IDA. Moreover, the proposed immunosensor system with the 3D IDA successfully worked for clinical analysis as shown by the sensitive detection of cardiac troponin I in human serum down to 100 fg/mL.

  5. Amplified impedimetric immunosensor based on instant catalyst for sensitive determination of ochratoxin A.

    Science.gov (United States)

    Tang, Juan; Huang, Yapei; Zhang, Cengceng; Liu, Huiqiong; Tang, Dianping

    2016-12-15

    A new impedimetric immunosensor for the fast determination of ochratoxin A (OTA) in food samples was developed based on the instant catalyst as enhancer. Initially, the signal tags were prepared via co-immobilization of anti-OTA antibody and amine-terminated dendrimer (PAMAM) on the graphene oxide nanosheets through the covalent interaction, which were utilized as a good platform for combining manganese ion (anti-OTA-GO-PAMAM-Mn(2+)). Upon target OTA introduction, a competitive-type immunoreaction was implemented between the analyte and the immobilized OTA-BSA on the electrode for the anti-OTA antibody on the graphene oxide nanosheets labels. After a competitive immunoassay format, the anti-OTA-GO-PAMAM-Mn(2+) were captured onto the electrode surface, which could induce the in situ formation of MnO2via classical redox reaction between Mn(2+) and KMnO4 on the immunesensing platform. Moreover, the generated MnO2 nanoparticles act as efficient catalyst could catalyze the 4-chloro-1-naphthol (4-CN) oxidation without H2O2 to generate an insoluble precipitation on the platform. Under the optimal conditions, the instant catalyst based impedimetric immunosensor displayed a wide dynamic working range between 0.1pgmL(-1) and 30ngmL(-1). The detection limit (LOD) of the assay was 0.055pgmL(-1). The developed method exhibited high selectivity and can be used for the determination of OTA in real red wine samples.

  6. Novel multilayered porous silicon-based immunosensor for determining Hydroxysafflor yellow A

    Science.gov (United States)

    Lv, Xiaoyi; Mo, Jiaqing; Jiang, Tao; Zhong, Furu; Jia, Zhenhong; Li, Jiangwei; Zhang, Fuchun

    2011-01-01

    External random factors have a great influence on the fabrication of accurate photonic crystal, especially porous silicon-based photonic crystals. Compared with the binary photonic crystal, polybasic structure photonic crystal shows more stability and smaller effect of the random fluctuation. In this paper, we have fabricated a novel simple porous silicon polybasic Bragg's mirror combined with excellent specific antigen-antibody inmunoreaction as an immunosensor for determining Hydroxysafflor yellow A (HSYA), which is the main chemical component of Carthamus tinctorius L. The binding of HSYA and the polyclonal anti-HSYA antibodies causes red shifts in the reflection spectrum of the sensor, and the red shift was proportional to the HSYA concentration with linear relationship ranging from 1 to 3 μg mL -1 with a detection limit of 0.78 ng mL -1. Importantly, this research offers hope for development of a commercial porous silicon-based immunosensor for component determination of C. tinctorius L. or other antigens.

  7. Novel multilayered porous silicon-based immunosensor for determining Hydroxysafflor yellow A

    Energy Technology Data Exchange (ETDEWEB)

    Lv Xiaoyi, E-mail: xiaoz813@sohu.com [School of Electronic and Information Engineering, Xi' an Jiaotong University, Xi' an 710049 (China); Mo Jiaqing [School of Electronic and Information Engineering, Xi' an Jiaotong University, Xi' an 710049 (China); Jiang Tao; Zhong Furu [College of Information Science and Engineering, Xinjiang University, Urumqi 830046 (China); Jia Zhenhong, E-mail: jzhh9009@sohu.com [College of Information Science and Engineering, Xinjiang University, Urumqi 830046 (China); Li Jiangwei; Zhang Fuchun [Key Laboratory of Xinjiang Biological Resources and Gene Engineering, College of Life Sciences and Technology, Xinjiang University, Urumqi 830046 (China)

    2011-01-01

    External random factors have a great influence on the fabrication of accurate photonic crystal, especially porous silicon-based photonic crystals. Compared with the binary photonic crystal, polybasic structure photonic crystal shows more stability and smaller effect of the random fluctuation. In this paper, we have fabricated a novel simple porous silicon polybasic Bragg's mirror combined with excellent specific antigen-antibody inmunoreaction as an immunosensor for determining Hydroxysafflor yellow A (HSYA), which is the main chemical component of Carthamus tinctorius L. The binding of HSYA and the polyclonal anti-HSYA antibodies causes red shifts in the reflection spectrum of the sensor, and the red shift was proportional to the HSYA concentration with linear relationship ranging from 1 to 3 {mu}g mL{sup -1} with a detection limit of 0.78 ng mL{sup -1}. Importantly, this research offers hope for development of a commercial porous silicon-based immunosensor for component determination of C. tinctorius L. or other antigens.

  8. Vesicle Motion during Sustained Exocytosis in Chromaffin Cells: Numerical Model Based on Amperometric Measurements.

    Science.gov (United States)

    Jarukanont, Daungruthai; Bonifas Arredondo, Imelda; Femat, Ricardo; Garcia, Martin E

    2015-01-01

    Chromaffin cells release catecholamines by exocytosis, a process that includes vesicle docking, priming and fusion. Although all these steps have been intensively studied, some aspects of their mechanisms, particularly those regarding vesicle transport to the active sites situated at the membrane, are still unclear. In this work, we show that it is possible to extract information on vesicle motion in Chromaffin cells from the combination of Langevin simulations and amperometric measurements. We developed a numerical model based on Langevin simulations of vesicle motion towards the cell membrane and on the statistical analysis of vesicle arrival times. We also performed amperometric experiments in bovine-adrenal Chromaffin cells under Ba2+ stimulation to capture neurotransmitter releases during sustained exocytosis. In the sustained phase, each amperometric peak can be related to a single release from a new vesicle arriving at the active site. The amperometric signal can then be mapped into a spike-series of release events. We normalized the spike-series resulting from the current peaks using a time-rescaling transformation, thus making signals coming from different cells comparable. We discuss why the obtained spike-series may contain information about the motion of all vesicles leading to release of catecholamines. We show that the release statistics in our experiments considerably deviate from Poisson processes. Moreover, the interspike-time probability is reasonably well described by two-parameter gamma distributions. In order to interpret this result we computed the vesicles' arrival statistics from our Langevin simulations. As expected, assuming purely diffusive vesicle motion we obtain Poisson statistics. However, if we assume that all vesicles are guided toward the membrane by an attractive harmonic potential, simulations also lead to gamma distributions of the interspike-time probability, in remarkably good agreement with experiment. We also show that

  9. Vesicle Motion during Sustained Exocytosis in Chromaffin Cells: Numerical Model Based on Amperometric Measurements.

    Directory of Open Access Journals (Sweden)

    Daungruthai Jarukanont

    Full Text Available Chromaffin cells release catecholamines by exocytosis, a process that includes vesicle docking, priming and fusion. Although all these steps have been intensively studied, some aspects of their mechanisms, particularly those regarding vesicle transport to the active sites situated at the membrane, are still unclear. In this work, we show that it is possible to extract information on vesicle motion in Chromaffin cells from the combination of Langevin simulations and amperometric measurements. We developed a numerical model based on Langevin simulations of vesicle motion towards the cell membrane and on the statistical analysis of vesicle arrival times. We also performed amperometric experiments in bovine-adrenal Chromaffin cells under Ba2+ stimulation to capture neurotransmitter releases during sustained exocytosis. In the sustained phase, each amperometric peak can be related to a single release from a new vesicle arriving at the active site. The amperometric signal can then be mapped into a spike-series of release events. We normalized the spike-series resulting from the current peaks using a time-rescaling transformation, thus making signals coming from different cells comparable. We discuss why the obtained spike-series may contain information about the motion of all vesicles leading to release of catecholamines. We show that the release statistics in our experiments considerably deviate from Poisson processes. Moreover, the interspike-time probability is reasonably well described by two-parameter gamma distributions. In order to interpret this result we computed the vesicles' arrival statistics from our Langevin simulations. As expected, assuming purely diffusive vesicle motion we obtain Poisson statistics. However, if we assume that all vesicles are guided toward the membrane by an attractive harmonic potential, simulations also lead to gamma distributions of the interspike-time probability, in remarkably good agreement with experiment. We

  10. A novel label-free microfluidic paper-based immunosensor for highly sensitive electrochemical detection of carcinoembryonic antigen.

    Science.gov (United States)

    Wang, Yang; Xu, Huiren; Luo, Jinping; Liu, Juntao; Wang, Li; Fan, Yan; Yan, Shi; Yang, Yue; Cai, Xinxia

    2016-09-15

    In this work, a highly sensitive label-free paper-based electrochemical immunosensor employing screen-printed working electrode (SPWE) for detection of carcinoembryonic antigen (CEA) was fabricated. In order to raise the detection sensitivity and immobilize anti-CEA, amino functional graphene (NH2-G)/thionine (Thi)/gold nanoparticles (AuNPs) nanocomposites were synthesized and coated on SPWE. The principle of the immunosensor determination was based on the fact that the decreased response currents of Thi were proportional to the concentrations of corresponding antigens due to the formation of antibody-antigen immunocomplex. Experimental results revealed that the immunoassay enabled the determination of standard CEA solutions with linear working ranges of 50pgmL(-1) to 500ngmL(-1), the limit of detections for CEA is 10pgmL(-1) (S/N=3) and its corresponding correlation coefficients were 0.996. Furthermore, the proposed immunosensor could be used for the determination of clinical serum samples. A large number of clinical serum samples were detected and the relative errors between measured values and reference concentrations were calculated. Results showed that this novel paper-based electrochemical immunosensor could provide a new platform for low cost, sensitive, specific, and point-of-care diagnosis in cancer detection.

  11. An ultrasensitive electrochemical immunosensor for apolipoprotein E4 based on fractal nanostructures and enzyme amplification.

    Science.gov (United States)

    Liu, Yibiao; Xu, Li-Ping; Wang, Shuqi; Yang, Weizhao; Wen, Yongqiang; Zhang, Xueji

    2015-09-15

    Human apolipoprotein E4 (APOE4) is a major risk factor for Alzheimer's disease (AD) and can greatly increase the morbidity. In this work, an ultrasensitive sandwich-type electrochemical immunosensor for the quantitative detection of APOE4 was designed based on fractal gold (FracAu) nanostructures and enzyme amplification. The FracAu nanostructures were directly electrodeposited by hydrogen tetrachloroaurate (HAuCl4) on polyelectrolytes modified indium tin oxide (ITO) electrode. The sensing performances of the modified interface were investigated by cyclic voltammetry (CV). After functionalization with HRP-labeled APOE4 antibody, the human APOE4 could be detected quantitatively by current response. The current response has a linear relationship with the logarithm of human APOE4 concentrations in a range from 1.0 to 10,000.0 ng/mL, with a detection limit of 0.3 ng/mL. The fabricated APOE4 electrochemical immunosensor exhibits strong specificity, high sensitivity, low detection limit and wide linear range. The detection of human APOE4 provides a strong support for the prevention of AD and early-stage warning for those susceptible populations.

  12. Simultaneous electrochemical immunosensor based on water-soluble polythiophene derivative and functionalized magnetic material.

    Science.gov (United States)

    Zhang, Xiaoyue; Ren, Xiang; Cao, Wei; Li, Yueyun; Du, Bin; Wei, Qin

    2014-10-03

    A novel, sensitive electrochemical immunosensor for simultaneous determination of squamous cell carcinoma associated antigen (SCC-Ag) and carcinoembryonic antigen (CEA) for the combined diagnosis of cervical cancer was designed. The amplification strategy for electrochemical immunoassay was based on poly[3-(1,1'-dimethyl-4-piperidine-methylene) thiophene-2,5-diylchloride] (PDPMT-Cl) and functionalized mesoporous ferroferric oxide nanoparticles (Fe3O4 NPs). PDPMT-Cl dispersed in chitosan solution with enhanced electrical conductivity and solubility was used as matrices to immobilize the first antibodies. Different redox probes (thionine (Th) and ferrocenecarboxylic acid (Fca)) functionalized Fe3O4 NPs incubated with two kinds of secondary antibodies to fabricate the labels. Using an electrochemical analysis technique, two well-separated peaks were generated by Th and Fca, making the simultaneous detection of two analytes on the electrode possible. Under optimized conditions, this method showed wide linear ranges of three orders of magnitude with the detection limits of 4 pg mL(-1) and 5 pg mL(-1), respectively. The disposable immunosensor possessed excellent clinical value in cervical cancer screening as well as convenient point-of-care diagnostics.

  13. Signal Amplification by Enzymatic Reaction in an Immunosensor Based on Localized Surface Plasmon Resonance (LSPR

    Directory of Open Access Journals (Sweden)

    Yong-Beom Shin

    2010-03-01

    Full Text Available An enzymatic reaction was employed as a means to enhance the sensitivity of an immunosensor based on localized surface plasmon resonance (LSPR. The reaction occurs after intermolecular binding between an antigen and an antibody on gold nano-island (NI surfaces. For LSPR sensing, the gold NI surface was fabricated on glass substrates using vacuum evaporation and heat treatment. The interferon-g (IFN-g capture antibody was immobilized on the gold NIs, followed by binding of IFN-g to the antibody. Subsequently, a biotinylated antibody and a horseradish peroxidase (HRP conjugated with avidin were simultaneously introduced. A solution of 4-chloro-1-naphthol (4-CN was then used for precipitation; precipitation was the result of the enzymatic reaction catalyzed the HRP on gold NIs. The LSPR spectra were obtained after each binding process. Using this method, the enzyme-catalyzed precipitation reaction on the gold NI surface was found to effectively amplify the change in the signal of the LSPR immunosensor after intermolecular binding.

  14. A droplet-based microfluidic immunosensor for high efficiency melamine analysis.

    Science.gov (United States)

    Choi, Jae-Won; Min, Kyong-Mi; Hengoju, Sundar; Kim, Gil-Jung; Chang, Soo-Ik; deMello, Andrew J; Choo, Jaebum; Kim, Hak Yong

    2016-06-15

    We report a droplet-based microfluidic immunosensor for the rapid and accurate detection of melamine, an organic base that has been implicated in widescale adulteration of food products such as milk. Our melamine assay is based on the competitive reaction between native melamine and a melamine-fluorescein isothiocyanate (FITC) conjugate against an anti-hapten antibody. The adoption of fluorescence polarization, allows the quantification of melamine in a more direct and rapid manner than established heterogeneous methods based on liquid chromatography, mass spectrometry, and enzyme-linked immunosorbent assay (ELISA). The detection protocol provides a limit of detection of 300 ppb, which is below the maximum allowable melamine levels (2.5 ppm) defined by the U.S. Food and Drug Administration and the European Commission to a significant extent.

  15. Impedimetric Biosensors and Immunosensors

    Directory of Open Access Journals (Sweden)

    Mamas I. Prodromidis

    2007-12-01

    Full Text Available The development of methods targeting the direct monitoring of antibody-antigen interactions is particularly attractive. The design of label-free affinity-based probing concepts is the objective of much current research, at both academic and industrial levels, towards establishing alternative methods to the already existing ELISA-based immunoassays. Among these, Electrochemical Impedance Spectroscopy (EIS represents one of the most powerful methods, due to the ability of EIS-based sensors to be more easily integrated into multi-array or microprocessor- controlled diagnostic tools. During the last decade, EIS and the concept of biochemical capacitors have been widely used for probing various types of biomolecular interactions (immunosensors, DNA hybridization, protein-protein interactions. So far, impedimetric or capacitive immunosensors have been successfully applied at the academic level. However, no prototypes have been released into the market, since major fundamental issues still exist. Even though this fact has brought the reliability of impedimetric immunosensors into question, features associated with electrochemical approaches, namely the ability to be miniaturized, remote control of implanted sensors, low cost of electrode mass production, and cost effective instrumentation (without need of high-energy sources keep impedimetric sensors particularly attractive as compared to other approaches based on microbalances, surface plasmon resonance or ellipsometry. This lecture outlines the theoretical background of impedimetric immunosensors and presents different types of impedimetric biosensors as well as the instrumental approaches that have been so far proposed in the literature.

  16. Towards the development of a single-step immunosensor based on an electrochemical screen-printed electrode strip coupled with immunomagnetic beads.

    Science.gov (United States)

    Volpe, G; Sozzo, U; Piermarini, S; Delibato, E; Palleschi, G; Moscone, D

    2013-01-01

    This work investigates the behaviour of two alternative systems that model the crucial event involved in any ELISA test, i.e. the molecular recognition between an antigen and its specific antibody on a solid phase, and its measurement. Each approach is devised with the goal of making possible a single-step, separation and wash-free amperometric magneto-immunosensor. Magnetic particles (MBs) are used as support for the immobilization of rabbit IgGs that are recognized by the specific anti-rabbit IgG-HRP. The assay protocol is based on the use of a series of small "reservoirs" containing phosphate buffer, hydroquinone, anti-rabbit IgG-HRP and an appropriate amount of MB-rabbit IgG. After a brief incubation, the content of each "reservoir" is transferred to one of the wells of a 8-well magnetized-screen-printed electrode strip. The resulting MB-IgG-anti-IgG-HRP chain, is then concentrated on the working electrode surface for electrochemical measurement. Two different approaches to monitor this immunological reaction are investigated. The first one is based on the enzyme-channeling principle (ECP) and involves the use of a second enzyme, glucose oxidase (GOD), immobilized on the working electrode previously modified with Prussian Blue. Since the H(2)O(2) produced by GOD is the co-substrate of the HRP enzyme, glucose is added into the well and the current, generated by the residual H(2)O(2), is measured. The second, more direct, approach is performed without exploiting ECP (no GOD enzyme), by adding H(2)O(2) into the well and measuring the current generated by the HRP product on a pristine screen-printed electrode. Both approaches yielded a typical sigmoidal binding curve, illustrating the discrimination between the signal produced by the immuno-bound HRP concentrated on the electrode surface, and the background signal due to HRP in the bulk solution.

  17. SPR based immunosensor for detection of Legionella pneumophila in water samples

    Science.gov (United States)

    Enrico, De Lorenzis; Manera, Maria G.; Montagna, Giovanni; Cimaglia, Fabio; Chiesa, Maurizio; Poltronieri, Palmiro; Santino, Angelo; Rella, Roberto

    2013-05-01

    Detection of legionellae by water sampling is an important factor in epidemiological investigations of Legionnaires' disease and its prevention. To avoid labor-intensive problems with conventional methods, an alternative, highly sensitive and simple method is proposed for detecting L. pneumophila in aqueous samples. A compact Surface Plasmon Resonance (SPR) instrumentation prototype, provided with proper microfluidics tools, is built. The developed immunosensor is capable of dynamically following the binding between antigens and the corresponding antibody molecules immobilized on the SPR sensor surface. A proper immobilization strategy is used in this work that makes use of an important efficient step aimed at the orientation of antibodies onto the sensor surface. The feasibility of the integration of SPR-based biosensing setups with microfluidic technologies, resulting in a low-cost and portable biosensor is demonstrated.

  18. Rapid and Highly Sensitive Detection of Lead Ions in Drinking Water Based on a Strip Immunosensor

    Directory of Open Access Journals (Sweden)

    Chuanlai Xu

    2013-03-01

    Full Text Available In this study, we have first developed a rapid and sensitive strip immunosensor based on two heterogeneously-sized gold nanoparticles (Au NPs probes for the detection of trace lead ions in drinking water. The sensitivity was 4-fold higher than that of the conventional LFA under the optimized conditions. The visual limit of detection (LOD of the amplified method for qualitative detection lead ions was 2 ng/mL and the LOD for semi-quantitative detection could go down to 0.19 ng/mL using a scanning reader. The method suffered from no interference from other metal ions and could be used to detect trace lead ions in drinking water without sample enrichment. The recovery of the test samples ranged from 96% to 103%. As the detection method could be accomplished within 15 min, this method could be used as a potential tool for preliminary monitoring of lead contamination in drinking water.

  19. Rapid and highly sensitive detection of lead ions in drinking water based on a strip immunosensor.

    Science.gov (United States)

    Kuang, Hua; Xing, Changrui; Hao, Changlong; Liu, Liqiang; Wang, Libing; Xu, Chuanlai

    2013-03-28

    In this study, we have first developed a rapid and sensitive strip immunosensor based on two heterogeneously-sized gold nanoparticles (Au NPs) probes for the detection of trace lead ions in drinking water. The sensitivity was 4-fold higher than that of the conventional LFA under the optimized conditions. The visual limit of detection (LOD) of the amplified method for qualitative detection lead ions was 2 ng/mL and the LOD for semi-quantitative detection could go down to 0.19 ng/mL using a scanning reader. The method suffered from no interference from other metal ions and could be used to detect trace lead ions in drinking water without sample enrichment. The recovery of the test samples ranged from 96% to 103%. As the detection method could be accomplished within 15 min, this method could be used as a potential tool for preliminary monitoring of lead contamination in drinking water.

  20. Nanoparticle-based immunosensors and immunoassays for aflatoxins.

    Science.gov (United States)

    Wang, Xu; Niessner, Reinhard; Tang, Dianping; Knopp, Dietmar

    2016-03-17

    Aflatoxins are naturally existing mycotoxins produced mainly by Aspergillus flavus and Aspergillus parasiticus, present in a wide range of food and feed products. Because of their extremely high toxicity and carcinogenicity, strict control of maximum residue levels of aflatoxins in foodstuff is set by many countries. In daily routine, different chromatographic methods are used almost exclusively. As supplement, in several companies enzyme immunoassay-based sample testing as primary screening is performed. Recently, nanomaterials such as noble metal nanoparticles, magnetic particles, carbon nanomaterials, quantum dots, and silica nanomaterials are increasingly utilized for aflatoxin determination to improve the sensitivity and simplify the detection. They are employed either as supports for the immobilization of biomolecules or as electroactive or optical labels for signal transduction and amplification. Several nanoparticle-based electrochemical, piezoelectric, optical, and immunodipstick assays for aflatoxins have been developed. In this review, we summarize these recent advances and illustrate novel concepts and promising applications in the field of food safety.

  1. Nanoparticle-based immunosensors and immunoassays for aflatoxins

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Xu; Niessner, Reinhard [Institute of Hydrochemistry and Chair of Analytical Chemistry, Technische Universität München, Marchioninistrasse 17, D-81377 München (Germany); Tang, Dianping [Key Laboratory of Analysis and Detection for Food Safety, MOE & Fujian Province, Department of Chemistry, Fuzhou University, Fuzhou 350108 (China); Knopp, Dietmar, E-mail: dietmar.knopp@ch.tum.de [Institute of Hydrochemistry and Chair of Analytical Chemistry, Technische Universität München, Marchioninistrasse 17, D-81377 München (Germany)

    2016-03-17

    Aflatoxins are naturally existing mycotoxins produced mainly by Aspergillus flavus and Aspergillus parasiticus, present in a wide range of food and feed products. Because of their extremely high toxicity and carcinogenicity, strict control of maximum residue levels of aflatoxins in foodstuff is set by many countries. In daily routine, different chromatographic methods are used almost exclusively. As supplement, in several companies enzyme immunoassay-based sample testing as primary screening is performed. Recently, nanomaterials such as noble metal nanoparticles, magnetic particles, carbon nanomaterials, quantum dots, and silica nanomaterials are increasingly utilized for aflatoxin determination to improve the sensitivity and simplify the detection. They are employed either as supports for the immobilization of biomolecules or as electroactive or optical labels for signal transduction and amplification. Several nanoparticle-based electrochemical, piezoelectric, optical, and immunodipstick assays for aflatoxins have been developed. In this review, we summarize these recent advances and illustrate novel concepts and promising applications in the field of food safety. - Highlights: • Novel concepts and promising applications of nanoparticle-based immunological methods for the determination of aflatoxins. • Inclusion of most important nanomaterials and hybrid nanostructures. • Inclusion of electrochemical, optical and mass-sensitive biosensors as well as optical and immunochromatographic assays.

  2. Amperometric biosensors for glucose, lactate, and glycolate based on oxidases and redox-modified siloxane polymers

    Science.gov (United States)

    Hale, Paul D.; Inagaki, Toru; Lee, Hung Sui; Skotheim, Terje A.; Karan, Hiroko I.; Okamoto, Yoshi

    1989-06-01

    Amperometric biosensors based on flavin-containing oxidases undergo several steps which produce a measurable current that is related to the concentration of substrate. In the initial step, the substrate converts the oxidized flavin adenine dinucleotide (FAD) or flavin mononucleotide (FMN) into the reduced form FADH sub 2 or FMNH sub 2. Because these cofactors are located well within the enzyme molecule, direct electron transfer to the surface of a conventional electrode does not occur to a measurable degree. A common method of facilitating this electron transfer is to introduce oxygen into the system because it is the natural acceptor for the oxidases; the oxygen is reduced by the FADH sub 2 or FMNH sub 2 to hydrogen peroxide, which can then be detected electrochemically. The major drawback to this approach is the fact that oxidation of hydrogen peroxide requires a large overpotential, thus making these sensors susceptible to interference from electroactive species. To lower the necessary applied potential, several non-physiological redox couples have been employed to shuttle electrons between the flavin moieties and the electrode. The present paper describes the development of amperometric biosensors based on flavin-containing enzymes and a family of polymeric mediators.

  3. Electrochemical cortisol immunosensors based on sonochemically synthesized zinc oxide 1D nanorods and 2D nanoflakes.

    Science.gov (United States)

    Vabbina, Phani Kiran; Kaushik, Ajeet; Pokhrel, Nimesh; Bhansali, Shekhar; Pala, Nezih

    2015-01-15

    We report on label free, highly sensitive and selective electrochemical immunosensors based on one-dimensional 1D ZnO nanorods (ZnO-NRs) and two-dimensional 2D ZnO nanoflakes (ZnO-NFs) which were synthesized on Au-coated substrates using simple one step sonochemical approach. Selective detection of cortisol using cyclic voltammetry (CV) is achieved by immobilizing anti-cortisol antibody (Anti-C(ab)) on the ZnO nanostructures (NSs). 1D ZnO-NRs and 2D ZnO-NFs provide unique sensing advantages over bulk materials. While 1D-NSs boast a high surface area to volume ratio, 2D-NSs with large area in polarized (0001) plane and high surface charge density could promote higher Anti-C(ab) loading and thus better sensing performance. Beside large surface area, ZnO-NSs also exhibit higher chemical stability, high catalytic activity, and biocompatibility. TEM studies showed that both ZnO-NSs are single crystalline oriented in (0001) plane. The measured sensing parameters are in the physiological range with a sensitivity of 11.86 µA/M exhibited by ZnO-NRs and 7.74 µA/M by ZnO-NFs with the lowest detection limit of 1 pM which is 100 times better than conventional enzyme-linked immunosorbant immunoassay (ELISA). ZnO-NSs based cortisol immunosensors were tested on human saliva samples and the performance were validated with conventional (ELISA) method which exhibits a remarkable correlation. The developed sensors can be integrated with microfluidic system and miniaturized potentiostat for point-of-care cortisol detection and such developed protocol can be used in personalized health monitoring/diagnostic.

  4. Disposable immunosensors for C-reactive protein based on carbon nanotubes field effect transistors.

    Science.gov (United States)

    Justino, Celine I L; Freitas, Ana C; Amaral, José P; Rocha-Santos, Teresa A P; Cardoso, Susana; Duarte, Armando C

    2013-04-15

    Label-free immunosensors based on single-walled carbon nanotubes field effect transistor (NTFET) devices were developed for the detection of C-reactive protein (CRP) which is currently the best validated inflammatory biomarker associated with cardiovascular diseases. The immunoreaction principle consists in the direct adsorption of CRP specific antibodies (anti-CRP) to single-walled carbon nanotubes (SWCNTs) networks. Such anti-CRP are the molecular receptors of CRP antigens which, in turn, can be detected by the developed NTFET devices in a linear dynamic range of 10(-4)-10(2) μg/mL. Thus, typical values of CRP (in blood serum) for healthy persons (5 μg/mL) corresponding to pathological states, can be both detected with the NTFET immunosensors, becoming an advantageous alternative as the basis for the development of analytical instrumentation for assessment of risk of occurrence of cardiovascular diseases. A log-log linear regression was applied to the experimental data with a correlation coefficient of r=0.9962 (pdevices (p=0.9582), demonstrating acceptable reproducibility. According to the experimental results, the estimate of detection limit (LOD, 10(-4)μg/mL) is 3-fold lower than that of some conventional immunoassay techniques for blood serum (e.g., LOD of 0.2 μg/mL for high-sensitivity enzyme-linked immunosorbent assay), and the dynamic range (10(-4)-10(2)μg/mL) is about 6-fold higher. Furthermore, this simple and low-cost methodology allows the use of sample volumes as low as 1 μL for the label-free detection of CRP.

  5. Amperometric Biosensor for Hydrogen Peroxide Based on Electrodeposited Sub-micrometer Gold Modified Glassy Carbon Electrode

    Institute of Scientific and Technical Information of China (English)

    WANG,Shu-Qing(王树青); CHEN,Jun(陈峻); LIN,Xiang-Qin(林祥钦)

    2004-01-01

    A new type of hydrogen peroxide amperometric biosensor was fabricated based on electrochemically deposited sub-micrometer Au particles(sm-Au)on a glassy carbon electrode(GCE).Electrochemical deposition condition was optimized for obtaining uniformly distributed sub-micrometer sized Au array on the electrode surface.The hydrogen peroxide sensor was fabricated by adsorbing phenothiazine methylene blue(MB)molecules on the surface of sm-Au and covering a cross-linked horseradish peroxidase(HRP)layer,labeled as HRP/MB/sm-Au/GCE.The characteristics of this biosensor were evaluated with respect to applied potential and pH.The amperometric response of the sensor was linear to the H2O2 concentration over a wide range of 9.9×10-6-1.11×10-2 mol/L.A detection limit(s/n=3)of 3.0×10-6 mol/L H2O2 was estimated for a sampled chronoamperometric detection at 1.5 min after potential step of 200 to-400 mV vs.SCE.The immobilized MB molecules shuttled electrons at(=0.77 and an apparent electron transfer rate constant of =0.053 s-1.Interference of ascorbic acid,dopamine and uric acid was investigated.This sensor has very good stability and reproducibility for long-term use.

  6. Development of urease based amperometric biosensors for the inhibitive determination of Hg (II).

    Science.gov (United States)

    Domínguez-Renedo, O; Alonso-Lomillo, M A; Ferreira-Gonçalves, L; Arcos-Martínez, M J

    2009-10-15

    Enzymatic amperometric procedures for measurement of Hg (II), based on the inhibitive action of this metal on urease enzyme activity, were developed. Screen-printed carbon electrodes (SPCEs) and gold nanoparticles modified screen-printed carbon electrodes (AuNPs/SPCEs) were used as supports for the cross-linking inmobilization of the enzyme urease. The amperometric response of urea was affected by the presence of Hg (II) ions which caused a decreasing in the current intensity. The optimum working conditions were found using experimental design methodology. Under these conditions, repeatability and reproducibility for both types of biosensors were determined, reaching values below 6% in terms of residual standard deviation. The detection limit obtained for Hg (II) was 4.2x10(-6)M for urease/SPCE biosensor and 5.6x10(-8)M for urease/AuNPs/SPCE biosensor. Analysis of the possible effect of the presence of foreign ions in the solution was performed. The method was applied to determine levels of Hg (II) in spiked human plasma samples.

  7. Fabrication of amperometric xanthine biosensors based on direct chemistry of xanthine oxidase

    Energy Technology Data Exchange (ETDEWEB)

    Gao Yansheng; Shen Chunping [Department of Chemistry, Soochow University, Suzhou, Jiangsu 215123 (China); Di Junwei, E-mail: djw@suda.edu.cn [Department of Chemistry, Soochow University, Suzhou, Jiangsu 215123 (China); Tu Yifeng [Department of Chemistry, Soochow University, Suzhou, Jiangsu 215123 (China)

    2009-08-31

    The construction of amperometric xanthine biosensor by immobilization of xanthine oxidase (XOD) on the multi-wall carbon nanotubes (CNTs) modified glassy carbon (GC) electrode surface was investigated. The direct chemistry of XOD was accomplished and the formal potential was about - 0.465 V (vs SCE). The heterogeneous electron transfer rate constant was evaluated to be 2.0 {+-} 0.3 s{sup -1}. The xanthine biosensor based on XOD entrapped in silica sol-gel (SG) thin film on CNTs-modified GC electrode surface was also investigated. The XOD still maintains its activity to xanthine. The amperometric response to xanthine showed a linear relation in the range from 0.2 {mu}M to 10 {mu}M and a detection limit of 0.1 {mu}M (S/N = 3). The enzyme electrode retained 95% of its initial activity after 90 days of storage. The sensor exhibited high sensitivity, rapid response and good long-term stability.

  8. Amperometric urea biosensors based on sulfonated graphene/polyaniline nanocomposite

    Directory of Open Access Journals (Sweden)

    Das G

    2015-08-01

    Full Text Available Gautam Das, Hyon Hee Yoon Department of Chemical and Biological Engineering, Gachon University, Seongnam, Gyeonggi-do, South Korea Abstract: An electrochemical biosensor based on sulfonated graphene/polyaniline nanocomposite was developed for urea analysis. Oxidative polymerization of aniline in the presence of sulfonated graphene oxide was carried out by electrochemical methods in an aqueous environment. The structural properties of the nanocomposite were characterized by Fourier-transform infrared, Raman spectroscopy, X-ray photoelectron spectroscopy, and scanning electron microscopy techniques. The urease enzyme-immobilized sulfonated graphene/polyaniline nanocomposite film showed impressive performance in the electroanalytical detection of urea with a detection limit of 0.050 mM and a sensitivity of 0.85 µA·cm-2·mM-1. The biosensor achieved a broad linear range of detection (0.12–12.3 mM with a notable response time of approximately 5 seconds. Moreover, the fabricated biosensor retained 81% of its initial activity (based on sensitivity after 15 days of storage at 4°C. The ease of fabrication coupled with the low cost and good electrochemical performance of this system holds potential for the development of solid-state biosensors for urea detection. Keywords: electrochemical deposition, sulfonated graphene oxide, urease

  9. Amperometric urea biosensors based on sulfonated graphene/polyaniline nanocomposite.

    Science.gov (United States)

    Das, Gautam; Yoon, Hyon Hee

    2015-01-01

    An electrochemical biosensor based on sulfonated graphene/polyaniline nanocomposite was developed for urea analysis. Oxidative polymerization of aniline in the presence of sulfonated graphene oxide was carried out by electrochemical methods in an aqueous environment. The structural properties of the nanocomposite were characterized by Fourier-transform infrared, Raman spectroscopy, X-ray photoelectron spectroscopy, and scanning electron microscopy techniques. The urease enzyme-immobilized sulfonated graphene/polyaniline nanocomposite film showed impressive performance in the electroanalytical detection of urea with a detection limit of 0.050 mM and a sensitivity of 0.85 (μA · cm(-2)·mM(-1). The biosensor achieved a broad linear range of detection (0.12-12.3 mM) with a notable response time of approximately 5 seconds. Moreover, the fabricated biosensor retained 81% of its initial activity (based on sensitivity) after 15 days of storage at 4°C. The ease of fabrication coupled with the low cost and good electrochemical performance of this system holds potential for the development of solid-state biosensors for urea detection.

  10. Amperometric urea biosensors based on sulfonated graphene/polyaniline nanocomposite

    Science.gov (United States)

    Das, Gautam; Yoon, Hyon Hee

    2015-01-01

    An electrochemical biosensor based on sulfonated graphene/polyaniline nanocomposite was developed for urea analysis. Oxidative polymerization of aniline in the presence of sulfonated graphene oxide was carried out by electrochemical methods in an aqueous environment. The structural properties of the nanocomposite were characterized by Fourier-transform infrared, Raman spectroscopy, X-ray photoelectron spectroscopy, and scanning electron microscopy techniques. The urease enzyme-immobilized sulfonated graphene/polyaniline nanocomposite film showed impressive performance in the electroanalytical detection of urea with a detection limit of 0.050 mM and a sensitivity of 0.85 (μA · cm−2·mM−1. The biosensor achieved a broad linear range of detection (0.12–12.3 mM) with a notable response time of approximately 5 seconds. Moreover, the fabricated biosensor retained 81% of its initial activity (based on sensitivity) after 15 days of storage at 4°C. The ease of fabrication coupled with the low cost and good electrochemical performance of this system holds potential for the development of solid-state biosensors for urea detection. PMID:26346240

  11. Disposable amperometric biosensor based on nanostructured bacteriophages for glucose detection

    Science.gov (United States)

    Kang, Yu Ri; Hwang, Kyung Hoon; Kim, Ju Hwan; Nam, Chang Hoon; Kim, Soo Won

    2010-10-01

    The selection of electrode material profoundly influences biosensor science and engineering, as it heavily influences biosensor sensitivity. Here we propose a novel electrochemical detection method using a working electrode consisting of bio-nanowires from genetically modified filamentous phages and nanoparticles. fd-tet p8MMM filamentous phages displaying a three-methionine (MMM) peptide on the major coat protein pVIII (designated p8MMM phages) were immobilized on the active area of an electrochemical sensor through physical adsorption and chemical bonding. Bio-nanowires composed of p8MMM phages and silver nanoparticles facilitated sensitive, rapid and selective detection of particular molecules. We explored whether the composite electrode with bio-nanowires was an effective platform to detect the glucose oxidase. The current response of the bio-nanowire sensor was high at various glucose concentrations (0.1 µm-0.1 mM). This method provides a considerable advantage to demonstrate analyte detection over low concentration ranges. Especially, phage-enabled bio-nanowires can serve as receptors with high affinity and specificity for the detection of particular biomolecules and provide a convenient platform for designing site-directed multifunctional scaffolds based on bacteriophages and may serve as a simple method for label-free detection.

  12. Micro-flow Immunosensor Based on Thin-film Interdigitated Gold Array Microelectrodes for Cancer Biomarker Detection.

    Science.gov (United States)

    Ravalli, Andrea; Lozzi, Luca; Marrazza, Giovanna

    2016-01-01

    In this paper, we reported the development of a micro-flow label-free impedimetric biosensor based on the use of thin-film interdigitated gold array microelectrodes (IDA) for the detection of carbohydrate antigen 125 (CA125). The immunosensor is developed through the electropolymerization of anthranilic acid (AA) on the surface of IDA electrodes followed by the covalent attachment of anti-CA125 monoclonal antibody. CA125 protein affinity reaction was then evaluated by means of electrochemical impedance spectroscopy (EIS). The sensor was characterized by electrochemical techniques and scanning electron microscopy (SEM). Using the optimized experimental conditions, the developed immunosensor showed a good analytical performance for CA125 detection from 0 to 100 U/mL with estimated limit of detection (LOD = 3Sblank/Slope) of 7 U/mL.

  13. Amperometric biosensors for glucose, lactate, and glycolate based on oxidases and redox-modified siloxane polymers

    Energy Technology Data Exchange (ETDEWEB)

    Hale, P.D.; Inagaki, Toru; Lee, Hung Sui; Skotheim, T.A.; Karan, Hiroko I.; Okamoto, Yoshi (Brookhaven National Lab., Upton, NY (USA); Medgar Evers Coll., Brooklyn, NY (USA). Div. of Natural Science and Mathematics; Polytechnic Univ., Brooklyn, NY (USA). Dept. of Chemistry)

    1989-01-01

    Amperometric biosensors based on flavin-containing oxidases undergo several steps which produce a measurable current that is related to the concentration of substrate. In the initial step, the substrate converts the oxidized flavin adenine dinucleotide (FAD) or flavin mononucleotide (FMN) into the reduced form (FADH{sub 2} or FMNH{sub 2}). Because these cofactors are located well within the enzyme molecule, direct electron transfer to the surface of a conventional electrode does not occur to a measurable degree. A common method of facilitating this electron transfer is to introduce oxygen into the system because it is the natural acceptor for the oxidases; the oxygen is reduced by the FADH{sub 2} or FMNH{sub 2} to hydrogen peroxide, which can then be detected electrochemically. The major drawback to this approach is the fact that oxidation of hydrogen peroxide requires a large overpotential, thus making these sensors susceptible to interference from electroactive species. To lower the necessary applied potential, several non-physiological redox couples have been employed to shuttle electrons between the flavin moieties and the electrode. For example, sensors based on the ferrocene/ferricinium redox couple and on electrodes consisting of conducting salts such as TTF-TCNQ (tetrathiafulvalene-tetracyanoquinodimethane) have previously been reported. Electron relays have also been attached directly to the enzyme molecule to facilitate electron transfer. More recently, these studies have been extended to include systems where the mediating redox species are covalently attached to polymers such as poly(pyrrole), poly(vinylpyridine), and poly(siloxane). The present paper describes the development of amperometric biosensors based on flavin-containing enzymes and this latter family of polymeric mediators. 9 refs., 5 figs.

  14. Detection of vibrio cholerae O1 by using cerium oxide nanowires - based immunosensor with different antibody immobilization methods

    Science.gov (United States)

    Tam, Phuong Dinh; Hoang, Nguyen Luong; Lan, Hoang; Vuong, Pham Hung; Anh, Ta Thi Nhat; Huy, Tran Quang; Thuy, Nguyen Thanh

    2016-05-01

    In this work, we evaluated the effects of different antibody immobilization strategies on the response of a CeO2-nanowires (NWs)-based immunosensor for Vibrio cholerae O1 detection. Accordingly, the changes in the electron-transfer resistance ( R et ) from before to after cells bind to an antibody-modified electrode prepared by using three different methods of antibody immobilization were determined. The values were 16.2%, 8.3%, and 6.65% for the method that utilized protein A, antibodies activated by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC)/N-hydroxysuccinimide (NHS), and absorption, respectively. Cyclic voltammetry confirmed that the change in the current was highest for the immunosensors prepared using protein A (11%), followed by those prepared with EDC/NHS-activated antibodies (9%), and finally, those prepared through absorption (7.5%). The order of the antibody immobilization strategies in terms of resulting immunosensor detection limit and sensitivity was as follows order: absorption (3.2 × 103 CFU/mL; 45.1 Ω/CFU·mL-1) < EDC/NHS-activated antibody (1.0 × 103 CFU/mL; 50.6 Ω/CFU·mL-1) < protein A (1.0 × 102 CFU/mL; 65.8 Ω/CFU·mL-1). Thus, we confirmed that the protein A - mediated method showed significantly high cell binding efficiencies compared to the random immobilization method.

  15. Facile preparation of disposable immunosensor for Shigella flexneri based on multi-wall carbon nanotubes/chitosan composite

    Energy Technology Data Exchange (ETDEWEB)

    Zhao Guangying, E-mail: zhaogy-user@163.co [Food Safety Key Lab of Zhejiang Province, Department of Food Quality and Safety, Zhejiang Gongshang University, 149, Jiaogong Road, Hangzhou 310035, Zhejiang Province (China); Zhan Xuejia [Food Safety Key Lab of Zhejiang Province, Department of Food Quality and Safety, Zhejiang Gongshang University, 149, Jiaogong Road, Hangzhou 310035, Zhejiang Province (China)

    2010-02-28

    Based on multi-wall carbon nanotubes (MWCNT)/chitosan/horseradish peroxidase labeled antibodies to Shigella flexneri (HRP-anti-S. flexneri) biocomposite film on a screen-printed electrode (SPE) surface, a disposable immunosensor has been developed for the rapid detection of S. flexneri. The HRP-anti-S. flexneri can be entrapped into MWCNT/chitosan composite matrix without other cross-linking agent. Thionine and H{sub 2}O{sub 2} were used as the mediator and substrate, respectively. The surface morphologies of modified films were characterized by atomic force microscope (AFM). Cyclic voltammery (CV) was carried out to characterize the electrochemical properties of the immobilization of materials on the electrode surface and quantified S. flexneri. Due to the strong electrocatalytic properties of MWCNT and HRP toward H{sub 2}O{sub 2}, the response signal was significantly amplified. S. flexneri could be detected by the decrease of the reduction peak current before and after immunoreaction. Under optimal conditions, S. flexneri could be detected in the range of 10{sup 4} to 10{sup 10} cfu mL{sup -1}, with a detection limit of 2.3 x 10{sup 3} cfu mL{sup -1} (S/N = 3). Furthermore, the proposed immunosensor exhibited a satisfactory specificity, reproducibility, stability and accuracy, indicating that the proposed immunosensor has potential application for a facile, rapid and harmless immunoassay.

  16. A photoelectrochemical immunosensor for detection of α-fetoprotein based on Au-ZnO flower-rod heterostructures

    Science.gov (United States)

    Han, Zhizhong; Luo, Min; Chen, Li; Chen, Jinghua; Li, Chunyan

    2017-04-01

    In this work, a novel label free photoelectrochemical (PEC) immunosensor has been developed for the detection of α-fetoprotein (AFP). The immunosensor was based on Au-ZnO flower-rods (FRs) heterostructure, where Au nanoparticles (NPs) were firstly electrodeposited by cyclic voltammetry methods. Scanning electron microscopy (SEM), X-ray diffraction (XRD), Mott-Schottky plot (MS), UV-vis diffuse reflectance spectrum and fluorescence emission spectrum were used for the characterizations of Au-ZnO FRs. The results demonstrated that Au NPs not only obviously enhanced the visible light absorption of ZnO FRs due to surface plasmon resonance (SPR) but also improved the separation of photo-generated electron-hole pairs. Therefore, the photocurrent of Au-ZnO FRs was increased under simulated sunlight. The photocurrent was reduced after the specific antibody-antigen immune reaction. And the photocurrent decrement was linear with the logarithm of AFP antigen concentration in the range from 0.005 ng mL-1 to 50 ng mL-1 with a low detection limit of 0.56 pg mL-1 (S/N = 3). The PEC immunosensor also exhibited high anti-interference property and acceptable stability. This work would provide a promising photoelectrochemical strategy for the detection of other proteins in clinical diagnosis.

  17. Faraday cage-type electrochemiluminescence immunosensor for ultrasensitive detection of Vibrio vulnificus based on multi-functionalized graphene oxide.

    Science.gov (United States)

    Guo, Zhiyong; Sha, Yuhong; Hu, Yufang; Yu, Zhongqing; Tao, Yingying; Wu, Yanjie; Zeng, Min; Wang, Sui; Li, Xing; Zhou, Jun; Su, Xiurong

    2016-10-01

    A novel Faraday cage-type electrochemiluminescence (ECL) immunosensor devoted to the detection of Vibrio vulnificus (VV) was fabricated. The sensing strategy was presented by a unique Faraday cage-type immunocomplex based on immunomagnetic beads (IMBs) and multi-functionalized graphene oxide (GO) labeled with (2,2'-bipyridine)(5-aminophenanthroline)ruthenium (Ru-NH2). The multi-functionalized GO could sit on the electrode surface directly due to the large surface area, abundant functional groups, and good electronic transport property. It ensures that more Ru-NH2 is entirely caged and become "effective," thus improving sensitivity significantly, which resembles extending the outer Helmholtz plane (OHP) of the electrode. Under optimal conditions, the developed immunosensor achieves a limit of detection as low as 1 CFU/mL. Additionally, the proposed immunosensor with high sensitivity and selectivity can be used for the detection of real samples. The novel Faraday cage-type method has shown potential application for the diagnosis of VV and opens up a new avenue in ECL immunoassay. Graphical abstract Faraday cage-type immunoassay mode for ultrasensitive detection by extending OHP.

  18. A polyaniline based ultrasensitive potentiometric immunosensor for cardiac troponin complex detection.

    Science.gov (United States)

    Zhang, Qi; Prabhu, Alok; San, Avdar; Al-Sharab, Jafar F; Levon, Kalle

    2015-10-15

    An ultrasensitive immunosensor based on potentiometric ELISA for the detection of a cardiac biomarker, troponin I-T-C (Tn I-T-C) complex, was developed. The sensor fabrication involves typical sandwich ELISA procedures, while the final signal readout was achieved using open circuit potentiometry (OCP). Glassy carbon (GC) working electrodes were first coated with emulsion-polymerized polyaniline/dinonylnaphthalenesulfonic acid (PANI/DNNSA) and the coated surface was utilized as a transducer layer on which sandwich ELISA incubation steps were performed. An enzymatic reaction between o-phenylenediamine (OPD) and hydrogen peroxide (H2O2) was catalyzed by horseradish peroxidase (HRP) labeled on the secondary antibodies. The polymer transducer charged state was mediated through electron (e(-)) and charge transfers between the transducer and charged species generated by the same enzymatic reaction. Such a change in the polymer transducer led to potential variations against an Ag/AgCl reference electrode as a function of Tn I-T-C complex concentration during incubations. The sequence of OPD and H2O2 additions, electrochemical properties of the PANI/DNNSA layer and non-specific binding prevention were all crucial factors for the assay performance. Under optimized conditions, the assay has a low limit of detection (LOD) ( 6 orders of magnitude), high repeatability (coefficient of variance < 8% for all concentrations higher than 5 pg/mL) and a short detection time (< 10 min).

  19. Electrochemical Immunosensor for a-Fetoprotein Based on Gold Nanoparticles/Graphene-Prussian Blue

    Institute of Scientific and Technical Information of China (English)

    张戈; 黄吴; 王伦; 王广凤

    2012-01-01

    The electrochemical immunosensor for a-fetoprotein (AFP) was fabricated based on the platform of gold nanoparticles (GNP)/graphene (Gr)-prussian blue (PB). By electrodeposition, GNP were modified on the surface of the prepared Gr-PB. The anti-AFP-1, l'-ferrocenedicarboxylic acid (FcDA) as label was directly immobilized on the platform of GNP/Gr-PB. And after the immunoreactions, the formed complex inhibited the electron transfer and decreased the catalytic current of FcDA toward the reduction of H2O2. And in the range of 10-3200 pgomL^-1, the decreased current is linear with the concentration of AFP, with a detection limit of 3 pg.mL-1. The developed im- munoassay method showed good precision, high sensitivity, acceptable stability and reproducibility, and could be used for the detection of real samples with consistent results in comparison with those obtained by the enzyme linked immunosorbent assay (ELISA) method.

  20. Flow-Injection Amperometric Determination of Tacrine based on Ion Transfer across a Water–Plasticized Polymeric Membrane Interface

    OpenAIRE

    Rueda, C.; Joaquin A. Ortuño

    2007-01-01

    A flow-injection pulse amperometric method for determining tacrine, based on ion transfer across a plasticized poly(vinyl chloride) (PVC) membrane, was developed. A four-electrode potentiostat with ohmic drop compensation was used, while a flow-through cell incorporated the four electrodes and the membrane, which contained tetrabutylammonium tetraphenylborate. The influence of the applied potential and of the flow-injection variables on the determination of tacrine was studied. In the selecte...

  1. Field-based detection and monitoring of uranium in contaminated groundwater using two immunosensors

    Energy Technology Data Exchange (ETDEWEB)

    Melton, S.J.; Yu, H.; Williams, K.H.; Morris, S.A.; Long, P.E.; Blake, D.A.

    2009-05-01

    Field-based monitoring of environmental contaminants has long been a need for environmental scientists. Described herein are two kinetic exclusion-based immunosensors, a field portable sensor (FPS) and an inline senor, that were deployed at the Integrated Field Research Challenge Site of the U.S. Department of Energy in Rifle, CO. Both sensors utilized a monoclonal antibody that binds to a U(VI)-dicarboxyphenanthroline complex (DCP) in a kinetic exclusion immunoassay format. These sensors were able to monitor changes of uranium in groundwater samples from {approx} 1 {micro}M to below the regulated drinking water limit of 126 nM (30 ppb). The FPS is a battery-operated sensor platform that can determine the uranium level in a single sample in 5-10 min, if the instrument has been previously calibrated with standards. The average minimum detection level (MDL) in this assay was 0.33 nM (79 ppt), and the MDL in the sample (based on a 1:200?1:400 dilution) was 66?132 nM (15.7?31.4 ppb). The inline sensor, while requiring a grounded power source, has the ability to autonomously analyze multiple samples in a single experiment. The average MDL in this assay was 0.12 nM (29 ppt), and the MDL in the samples (based on 1:200 or 1:400 dilutions) was 24?48 nM (5.7?11.4 ppb). Both sensor platforms showed an acceptable level of agreement (r{sup 2} = 0.94 and 0.76, for the inline and FPS, respectively) with conventional methods for uranium quantification.

  2. Electrochemical immunosensor based on nanoporpus gold loading thionine for carcinoembryonic antigen.

    Science.gov (United States)

    Sun, Xiaobin; Ma, Zhanfang

    2013-05-30

    Nanoporous gold (NPG) has recently received considerable attention in analytical electrochemistry because of its good conductivity and large specific surface area. A facile layer-by-layer assembly technique fabricated NPG was used to construct an electrochemical immunosensor for carcinoembryonic antigen (CEA). NPG was fabricated on glassy carbon (GC) electrode by alternatively assembling gold nanoparticles (AuNPs) and silver nanoparticles (AgNPs) using 1,4-benzenedimethanethiol as a cross-linker, and then AgNPs were dissolved with HNO3. The thionine was absorbed into the NPG and then gold nanostructure was electrodeposited on the surface through the electrochemical reduction of gold chloride tetrahydrate (HAuCl4). The anti-CEA was directly adsorbed on gold nanostructure fixed on the GC electrode. The linear range of the immunosensor was from 10 pg mL(-1) to 100 ng mL(-1) with a detection limit of 3 pg mL(-1) (S/N=3). The proposed immunosensor has high sensitivity, wide linear range, low detection limit, and good selectivity. The present method could be widely applied to construct other immunosensors.

  3. An Electrochemiluminescence Immunosensor Based on Gold-Magnetic Nanoparticles and Phage Displayed Antibodies.

    Science.gov (United States)

    Mu, Xihui; Tong, Zhaoyang; Huang, Qibin; Liu, Bing; Liu, Zhiwei; Hao, Lanqun; Dong, Hua; Zhang, Jinping; Gao, Chuan

    2016-02-27

    Using the multiple advantages of the ultra-highly sensitive electrochemiluminescence (ECL) technique, Staphylococcus protein A (SPA) functionalized gold-magnetic nanoparticles and phage displayed antibodies, and using gold-magnetic nanoparticles coated with SPA and coupled with a polyclonal antibody (pcAb) as magnetic capturing probes, and Ru(bpy)₃(2+)-labeled phage displayed antibody as a specific luminescence probe, this study reports a new way to detect ricin with a highly sensitive and specific ECL immunosensor and amplify specific detection signals. The linear detection range of the sensor was 0.0001~200 µg/L, and the limit of detection (LOD) was 0.0001 µg/L, which is 2500-fold lower than that of the conventional ELISA technique. The gold-magnetic nanoparticles, SPA and Ru(bpy)₃(2+)-labeled phage displayed antibody displayed different amplifying effects in the ECL immunosensor and can decrease LOD 3-fold, 3-fold and 20-fold, respectively, compared with the ECL immunosensors without one of the three effects. The integrated amplifying effect can decrease the LOD 180-fold. The immunosensor integrates the unique advantages of SPA-coated gold-magnetic nanoparticles that improve the activity of the functionalized capturing probe, and the amplifying effect of the Ru(bpy)₃(2+)-labeled phage displayed antibodies, so it increases specificity, interference-resistance and decreases LOD. It is proven to be well suited for the analysis of trace amounts of ricin in various environmental samples with high recovery ratios and reproducibility.

  4. An Electrochemiluminescence Immunosensor Based on Gold-Magnetic Nanoparticles and Phage Displayed Antibodies

    Directory of Open Access Journals (Sweden)

    Xihui Mu

    2016-02-01

    Full Text Available Using the multiple advantages of the ultra-highly sensitive electrochemiluminescence (ECL technique, Staphylococcus protein A (SPA functionalized gold-magnetic nanoparticles and phage displayed antibodies, and using gold-magnetic nanoparticles coated with SPA and coupled with a polyclonal antibody (pcAb as magnetic capturing probes, and Ru(bpy32+-labeled phage displayed antibody as a specific luminescence probe, this study reports a new way to detect ricin with a highly sensitive and specific ECL immunosensor and amplify specific detection signals. The linear detection range of the sensor was 0.0001~200 µg/L, and the limit of detection (LOD was 0.0001 µg/L, which is 2500-fold lower than that of the conventional ELISA technique. The gold-magnetic nanoparticles, SPA and Ru(bpy32+-labeled phage displayed antibody displayed different amplifying effects in the ECL immunosensor and can decrease LOD 3-fold, 3-fold and 20-fold, respectively, compared with the ECL immunosensors without one of the three effects. The integrated amplifying effect can decrease the LOD 180-fold. The immunosensor integrates the unique advantages of SPA-coated gold-magnetic nanoparticles that improve the activity of the functionalized capturing probe, and the amplifying effect of the Ru(bpy32+-labeled phage displayed antibodies, so it increases specificity, interference-resistance and decreases LOD. It is proven to be well suited for the analysis of trace amounts of ricin in various environmental samples with high recovery ratios and reproducibility.

  5. Amperometric hydrogen peroxide biosensor based on cobalt ferrite-chitosan nanocomposite

    Energy Technology Data Exchange (ETDEWEB)

    Yard Latin-Small-Letter-Dotless-I mc Latin-Small-Letter-Dotless-I , Feyza S.; Senel, Mehmet, E-mail: msenel@fatih.edu.tr; Baykal, Abduelhadi

    2012-02-01

    A novel H{sub 2}O{sub 2} biosensor based on horseradish peroxidase (HRP) immobilized into CoFe{sub 2}O{sub 4}-chitosan nanocomposite has been developed for the detection of hydrogen peroxide. The nanocomposite films were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). HRP has been entrapped into CoFe{sub 2}O{sub 4}-chitosan nanocomposite film and the immobilized enzyme could retain its bioactivity. This biosensor exhibited a fast amperometric response to hydrogen peroxide. The linear range for H{sub 2}O{sub 2} determination was from 3 Multiplication-Sign 10{sup -2} to 8 mM, with a detection limit of 2 Multiplication-Sign 10{sup -3} mM based on S/N = 3. The response time of the biosensor was 4 s. The effects of the pH and the temperature of the immobilized HRP electrode were also studied. - Highlights: Black-Right-Pointing-Pointer HRP biosensor based on CoFe{sub 2}O{sub 4}-chitosan nanocomposite has been developed for H{sub 2}O{sub 2} detection. Black-Right-Pointing-Pointer The biosensor seems to be simple to prepare, fast to respond, inexpensive and sensitive. Black-Right-Pointing-Pointer The biosensor had high sensitivity, good repeatability, reusability and long term stability.

  6. Quinone-Based Polymers for Label-Free and Reagentless Electrochemical Immunosensors: Application to Proteins, Antibodies and Pesticides Detection

    Directory of Open Access Journals (Sweden)

    Minh-Chau Pham

    2013-01-01

    Full Text Available Polyquinone derivatives are widely recognized in the literature for their remarkable properties, their biocompatibility, simple synthesis, and easy bio-functionalization. We have shown that polyquinones present very stable electroactivity in neutral aqueous medium within the cathodic potential domain avoiding side oxidation of interfering species. Besides, they can act as immobilized redox transducers for probing biomolecular interactions in sensors. Our group has been working on devices based on such modified electrodes with a view to applications for proteins, antibodies and organic pollutants using a reagentless label-free electrochemical immunosensor format. Herein, these developments are briefly reviewed and put into perspective.

  7. Amperometric inhibitive biosensor based on horseradish peroxidase-nanoporous gold for sulfide determination

    Science.gov (United States)

    Sun, Huihui; Liu, Zhuang; Wu, Chao; Xu, Ping; Wang, Xia

    2016-08-01

    As a well-known toxic pollutant, sulfide is harmful to human health. In this study, a simple and sensitive amperometric inhibitive biosensor was developed for the determination of sulfide in the environment. By immobilizing nanoporous gold (NPG) on glassy carbon electrode (GCE), and encapsulating horseradish peroxidase (HRP) onto NPG, a HRP/NPG/GCE bioelectrode for sulfide detection was successfully constructed based on the inhibition of sulfide on HRP activity with o-Phenylenediamine (OPD) as a substrate. The resulted HRP/NPG/GCE bioelectrode achieved a wide linear range of 0.1–40 μM in sulfide detection with a high sensitivity of 1720 μA mM‑1 cm‑2 and a low detection limit of 0.027 μM. Additionally, the inhibition of sulfide on HRP is competitive inhibition with OPD as a substrate by Michaelis-Menten analysis. Notably, the recovery of HRP activity was quickly achieved by washing the HRP/NPG/GCE bioelectrode using differential pulse voltammetry (DPV) technique in deaerated PBS (50 mM, pH 7.0) for only 60 s. Furthermore, the real sample analysis of sulfide by the HRP/NPG/GCE bioelectrode was achieved. Based on above results, the HRP/NPG/GCE bioelectrode could be a better choice for the real determination of sulfide compared to inhibitive biosensors previously reported.

  8. A novel sandwich electrochemiluminescence immunosensor for ultrasensitive detection of carbohydrate antigen 19-9 based on immobilizing luminol on Ag@BSA core/shell microspheres.

    Science.gov (United States)

    Zhang, Amin; Xiang, Hongkun; Zhang, Xin; Guo, Weiwei; Yuan, Enhui; Huang, Chusen; Jia, Nengqin

    2016-01-15

    A novel sandwich-type electrochemiluminescence immunosensor based on immobilizing luminol on Ag@BSA core/shell microspheres (Ag@BSA-luminol) for ultrasensitive detection of tumor marker carbohydrate antigen 19-9 (CA19-9) has been developed. Herein, magnetic carbon nanotubes (MAGCNTs) decorated with polyethylenimine (PEI) was used to construct the base of the immunosensor. MAGCNTs with prominent electrical conductivity and high surface area could be beneficial for promoting the electron transfer and loading plenty of primary antibodies (Ab1) via glutaraldehyde (GA). Meanwhile, the magnetic property of MAGCNTs makes it easy to be attached to the surface of magnetic glass carbon electrode (MGCE) through magnetism interaction, which provides an outstanding platform for this immunosensor. Moreover, Ag@BSA microspheres with large surface area, good stability, and excellent biocompatibility were desirable candidates for effective cross-link of CA19-9 detection antibodies (Ab2). A more interesting thing was that ELISA color reaction was used as an ultrasensitive strategy for identifying Ab2 was successfully coated on Ag@BSA with the naked eye. Additionally, we immobilized the luminol on the surface of Ag@BSA to prepare the target immunosensor. Immobilization of luminol on the surface of Ag@BSA could decrease the distance between luminophores and the electrode surface, leading to great enhancement of the ECL intensity of luminol in the present of hydrogen peroxide (H2O2). Under the optimal conditions, the intensity of the ECL immunosensor increased linearly with the logarithm of CA19-9 concentration in a wide linear range from 0.0005 to 150UmL(-1) with a detection limit of 0.0002UmL(-1) (S/N=3). All the results suggested the prepared CA19-9 immunosensor displayed high sensitivity, excellent stability and good specificity. The developed method opened a new avenue to clinical bioassay.

  9. Development of an amperometric biosensor based on peroxidases to quantify citrinin in rice samples.

    Science.gov (United States)

    Zachetti, Vanesa Gimena Lourdes; Granero, Adrian Marcelo; Robledo, Sebastián Noel; Zon, María Alicia; Fernández, Héctor

    2013-06-01

    An amperometric biosensor based on horseradish peroxidase (EC1.11.1.7,H2O2-oxide-reductases) to determine the content of citrinin mycotoxin in rice samples is proposed by the first time. The method uses carbon paste electrodes filled up with multi-walled carbon nanotubes embedded in a mineral oil, horseradish peroxidase, and ferrocene as a redox mediator. The biosensor is covered externally with a dialysis membrane, which is fixed to the body side of the electrode with a Teflon laboratory film, and an O-ring. The reproducibility and the repeatability were of 7.0% and 3.0%, respectively, showing a very good biosensor performance. The calibration curve was linear in a concentration range from 1 to 11.6nM. The limits of detection and quantification were 0.25nM and 0.75nM, respectively. For comparison, the citrinin content in rice samples was also determined by fluorimetric measurements. A very good correlation was obtained between the electrochemical and spectrophotometric methods.

  10. Amperometric Enzyme-based Gas Sensor for Formaldehyde: Impact of Possible Interferences

    Directory of Open Access Journals (Sweden)

    Ralf Moos

    2007-02-01

    Full Text Available In this work, cross-sensitivities and environmental influences on the sensitivityand the functionality of an enzyme-based amperometric sensor system for the directdetection of formaldehyde from the gas phase are studied. The sensor shows a linearresponse curve for formaldehyde in the tested range (0 - 15 vppm with a sensitivity of1.9 μA/ppm and a detection limit of about 130 ppb. Cross-sensitivities by environmentalgases like CO2, CO, NO, H2, and vapors of organic solvents like methanol and ethanol areevaluated as well as temperature and humidity influences on the sensor system. The sensorshowed neither significant signal to CO, H2, methanol or ethanol nor to variations in thehumidity of the test gas. As expected, temperature variations had the biggest influence onthe sensor sensitivity with variations in the sensor signal of up to 10 % of the signal for 5vppm CH2O in the range of 25 - 30 °C.

  11. Self-powered competitive immunosensor driven by biofuel cell based on hollow-channel paper analytical devices.

    Science.gov (United States)

    Li, Shuai; Wang, Yanhu; Ge, Shenguang; Yu, Jinghua; Yan, Mei

    2015-09-15

    A mediator-less and compartment-less glucose/O2 enzymatic biofuel cell (BFC) was introduced into microfluidic paper-based analytical devices (μ-PADs) that relies on flow in hollow channels with silver nanoparticles/graphene modified paper electrode as the anodic and cathodic substrate, to implement self-powered sensitive carcinoembryonic antigen (CEA) detection. Glucose dehydrogenase (GDH)-gold nanoparticles bioconjugate modified with CEA acted as a biocatalyst for enhancing glucose oxidation in the bioanode, as well as the transducing enzyme for signaling magnification. Similarly, nanoporous PtNi/bilirubin oxidase (BOD) acted as a biocatalyst for enhancing O2 reduction in the biocathode. With an increase in the concentration of CEA, the amount of CEA-Au-GDH bioconjugate on bioanode decreases, thus leading to the lower output of the as-prepared BFC. This proposed BFC-based self-powered immunosensor for CEA possessed largely increased linear detection range from 1 pg mL(-1) to 0.5 μg mL(-)(1) with a detection limit of 0.7 pg mL(-)(1). The proposed BFC-based self-powered immunosensor shows high sensitivity, stability, and reproducibility and can become a promising platform for other protein detection.

  12. Chemiluminescence multichannel immunosensor

    Energy Technology Data Exchange (ETDEWEB)

    Yacoub-George, E.; Scheithauer, W.; Koppi, A.; Meixner, L.; Drost, S. [Fraunhofer Inst. fuer Mikroelektronische Schaltungen und Systeme (IMS), Muenchen (Germany); Wolf, H. [Regensburg Univ. (Germany). Inst. of Medical Microbiology and Hygiene

    2001-07-01

    An automated chemiluminescence multichannel immunosensor for the parallel detection of three different substances is presented. The system is based on the chemiluminescence capillary ELISA technique in combination with a miniaturised fluidics system. Fused silica capillaries (FSC) are used as support for the immobilized antibodies. The sensing component consists of three stripe photodiodes. An integrated flow-through system ensures automation of the assay cycles. Data on the detection of the bacterial toxin staphylococcal enterotoxin B (SEB), the bacteriophage M13 and Escherichia coli O157:H7 are presented. (orig.)

  13. An Elegant Analysis of White Spot Syndrome Virus Using a Graphene Oxide/Methylene Blue based Electrochemical Immunosensor Platform

    Science.gov (United States)

    Natarajan, Anusha; Devi, K. S. Shalini; Raja, Sudhakaran; Senthil Kumar, Annamalai

    2017-01-01

    White spot syndrome virus (WSSV) is a major devastating virus in aquaculture industry. A sensitive and selective diagnostic method for WSSV is a pressing need for the early detection and protection of the aquaculture farms. Herein, we first report, a simple electrochemical immunosensor based on methylene blue dye (MB) immobilized graphene oxide modified glassy carbon electrode (GCE/GO@MB) for selective, quick (35 ± 5 mins) and raw sample analysis of WSSV. The immunosensor was prepared by sequential modification of primary antibody, blocking agent (bovine serum album), antigen (as vp28 protein), secondary antibody coupled with horseradish peroxidase (Ab2-HRP) on the GCE/GO@MB. The modified electrode showed a well-defined redox peak at an equilibrium potential (E1/2), −0.4 V vs Ag/AgCl and mediated H2O2 reduction reaction without any false positive result and dissolved oxygen interferences in pH 7 phosphate buffer solution. Under an optimal condition, constructed calibration plot was linear in a range of 1.36 × 10−3 to 1.36 × 107 copies μL−1 of vp28. It is about four orders higher sensitive than that of the values observed with polymerase chain reaction (PCR) and western blot based WSSV detection techniques. Direct electrochemical immunosensing of WSSV in raw tissue samples were successfully demonstrated as a real sample system. PMID:28393929

  14. Photoelectrochemical Immunosensor Array Based on Thioglycolic Acid Capped CdS Quantum Dots for Multiplexed Detection of Veterinary Drug Residues

    Institute of Scientific and Technical Information of China (English)

    肖飞; 赖彦君; 张苧丹; 白静; 鲜跃仲; 金利通

    2012-01-01

    A photoelectrochemical immunosensor based on multi-electrode array was developed for simultaneous and sen- sitive determination of veterinary drug residues. In this system, poly(dimethyldiallylammonium chloride) (PDDA), Au nanoparticles (Au NPs) and thioglycolic acid (TGA)-capped CdS quantum dots (QDs) were layer-by-layer as- sembled onto the home-made Au electrode array. The assembling process of the (CdS/PDDA/Au NPs/PDDA), mul- tilayer was characterized by electrochemical impedance spectroscopy. And then the antibodies for clenbuterol (CB), ractopamine (RAC) and chloramphenicol (CAP) were covalently immobilized onto the Au electrode array by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) coupling reaction, respectively. The concentrations of CB, RAC and CAP were measured based on the photoelectrochemical effects of CdS QDs. Under the optimal conditions the limits of detection (LOD) for CB, RAC and CAP were 25, 50 and 2.2 pg/mL (3a), respectively, with acceptable recovery over the range of 95.40%--105.5% in pig liver samples. All results indicate that the immunosensor array system has potential application for practical, effective and high throughput analysis of veterinary drugs residues.

  15. Experimental study of Love-wave immunosensors based on ZnO/LiTaO3 structures.

    Science.gov (United States)

    Zhou, Feng-mei; Li, Zhe; Fan, Li; Zhang, Shu-yi; Shui, Xiu-ji

    2010-03-01

    Experimental study of Love-mode immunosensors based on structures of ZnO/36 degrees YX-LiTaO3 is presented, in which the ZnO films with c-axis (002) orientation have been successfully grown on the 36 degrees YX-LiTaO3 substrates by RF magnetron sputtering technique. Then the Love-mode immunosensors based on the ZnO/36 degrees YX-LiTaO3 structures and monitoring antibody-antigen immunoreactions in aqueous solutions in real time are fabricated. The experimental results show that the optimal thickness of ZnO layers is about 1.20 microm in the structures deposited on 36 degrees YX-LiTaO3 substrates, which is much less than that of SiO2 overlayers about 6 microm. The antibody-antigen immunoreaction experiments also show that the frequency shifts of the sensors with 1.33 microm ZnO films are proportional to the concentration of antigen in solution as the concentration range less than 100 microg/ml.

  16. Screen Printed Carbon Electrode Based Electrochemical Immunosensor for the Detection of Dengue NS1 Antigen

    Directory of Open Access Journals (Sweden)

    Om Parkash

    2014-11-01

    Full Text Available An electrochemical immunosensor modified with the streptavidin/biotin system on screen printed carbon electrodes (SPCEs for the detection of the dengue NS1 antigen was developed in this study. Monoclonal anti-NS1 capture antibody was immobilized on streptavidin-modified SPCEs to increase the sensitivity of the assay. Subsequently, a direct sandwich enzyme linked immunosorbent assay (ELISA format was developed and optimized. An anti-NS1 detection antibody conjugated with horseradish peroxidase enzyme (HRP and 3,3,5,5'-tetramethybezidine dihydrochloride (TMB/H2O2 was used as an enzyme mediator. Electrochemical detection was conducted using the chronoamperometric technique, and electrochemical responses were generated at −200 mV reduction potential. The calibration curve of the immunosensor showed a linear response between 0.5 µg/mL and 2 µg/mL and a detection limit of 0.03 µg/mL. Incorporation of a streptavidin/biotin system resulted in a well-oriented antibody immobilization of the capture antibody and consequently enhanced the sensitivity of the assay. In conclusion, this immunosensor is a promising technology for the rapid and convenient detection of acute dengue infection in real serum samples.

  17. Preparation of Au-Pt nanostructures by combining top-down with bottom-up strategies and application in label-free electrochemical immunosensor for detection of NMP22.

    Science.gov (United States)

    Jia, Hongying; Gao, Picheng; Ma, Hongmin; Wu, Dan; Du, Bin; Wei, Qin

    2015-02-01

    A novel label-free amperometric immunosensor for sensitive detection of nuclear matrix protein 22 (NMP22) was developed based on Au-Pt bimetallic nanostructures, which were prepared by combining top-down with bottom-up strategies. Nanoporous gold (NPG) was prepared by "top-down" dealloying of commercial Au/Ag alloy film. After deposition of NPG on an electrode, Pt nanoparticles (PtNPs) were further decorated on NPG by "bottom-up" electrodeposition. The prepared bimetallic nanostructures combine the merits of both NPG and PtNPs, and show a high electrocatalytic activity towards the reduction of H2O2. The label-free immunosensor was constructed by directly immobilizing antibody of NMP22 (anti-NMP22) on the surface of bimetallic nanostructures. The immunoreaction induced amperometric response could be detected and negatively correlated to the concentration of NMP22. Bimetallic nanostructure morphologies and detection conditions were investigated to obtain the best sensing performance. Under the optimal conditions, a linear range from 0.01ng/mL to 10ng/mL and a detection limit of 3.33pg/mL were obtained. The proposed immunosensor showed high sensitivity, good selectivity, stability, reproducibility, and regeneration for the detection of NMP22, and it was evaluated in urine samples, receiving satisfactory results.

  18. A competitive photoelectrochemical immunosensor based on a CdS-induced signal amplification strategy for the ultrasensitive detection of dexamethasone.

    Science.gov (United States)

    Wang, Xueping; Yan, Tao; Li, Yan; Liu, Yixin; Du, Bin; Ma, Hongmin; Wei, Qin

    2015-12-09

    A novel photoelectrochemical immunosensor based on the competitive strategy is proposed for the specific detection of dexamethasone (DXM). Graphitic carbon nitride coupled with bismuth sulfide are used as the sensing matrix for the immobilization of BSA-DXM on the electrode surface, while cadmium sulfide functionalized titanium dioxide (TiO2@CdS) is used as the photoelectric active labels of anti-DXM. Due to the perfect matching of energy levels between TiO2 and CdS, the in situ prepared composite labels show excellent photocurrent response under visible lights. The competitive binding of DXM in sample solutions and BSA-DXM on the electrode surface reduces the specific attachment of labels to the electrode, resulting in a decrease of the photocurrent intensity. Greatly enhanced sensitivity is achieved after the optimization of the detection conditions. Under the optimal detection condition, the well-designed immunosensor for DXM exhibits a low detection limit of 2 pg ∙ mL(-1). Additionally, the proposed immunoassay system shows high specificity, good reproducibility and acceptable stability, which is also expected to become a promising platform for the detection of other small molecules.

  19. Dual immunosensor based on methylene blue-electroadsorbed graphene oxide for rapid detection of the influenza A virus antigen.

    Science.gov (United States)

    Veerapandian, Murugan; Hunter, Robert; Neethirajan, Suresh

    2016-08-01

    Rapid detection of influenza viral infections in poultry facilities is advantageous in several aspects such as environmental/personal safety, food-security, and socio-economy. Herein, we report the development of an electrochemical-based dual-sensor platform composed of methylene blue-electroadsorbed graphene oxide nanostructures modified with monoclonal antibodies against the HA proteins of H5N1 and H1N1. Bio-functional layers comprised of chitosan and protein-A molecules were implemented at the interface of the sensor element and antibodies, which synergistically enriched the bio-activity of immobilized antibodies for the immune complex formation. The differential pulse voltammetric signals resulted from the developed immunosensor platform exhibited a good correlation (R(2)=0.9978 for H1N1 and R(2)=0.9997 for H5N1) for the wide range of target concentrations 25-500pM). Chronoamperometric study also revealed the amplified current sensitivity of the immunoelectrodes even at the picomolar level. The proposed immunosensor design not only provides rapid analytical response time (<1min) but simplicity in fabrication and instrumentation, which paves an attractive platform for on-farm monitoring of viral infections.

  20. Amperometric Metronidazole Sensor Based on the Supermolecular Recognition by Metalloporphyrin Incorporated In Carbon Paste Electrode

    Directory of Open Access Journals (Sweden)

    Ru-Qin Yu

    2003-03-01

    Full Text Available An amperometric metronidazole (MTZ sensor using a glycosylated metalloporphyrin as a recognition element, which was incorporated in a carbon paste electrode, is reported. For the preparation of a MTZ-sensitive active material, 5, 10, 15, 20-tetrakis [2-(2, 3, 4, 6-tetraacetyl-β-D-glucopyranosyl-1-O-phenyl]porphyrin (T(oglu PPH2 and its Mn(III complex MnT(o-gluPPCl were synthesized from the reaction of pyrrole with ortho-acetylglycosylated benzaldehyde by Lindsay’s method. The MnT(oglu PPCl-modified electrode showed excellent selectivity toward MTZ with respect to a number of interferents and exhibited stable response. The calibration graph obtained with the proposed sensor was linear over the range of 2.9×10-3-5.8×10-8 M/L, with a detection limit of 5.8×10-8 M/L for MTZ. Cyclic voltammetric measurements indicated that MnT(oglu PPCl included in graphite-epoxy resin matrices could efficiently mediate electron transfer from the base electrode to MTZ causing a decrease of reduction potential for MTZ detection. The sensor could be regenerated by simply polishing with an alumina paper, with an excellent reproducibility (RSD=1.6%. The experimental conditions such as pH and applied working potential were optimized. The prepared sensor is applied for the determination of MTZ in pharmaceutical preparations and the results agreed with the values obtained by the pharmacopoeia method.

  1. Label-free Electrochemiluminescent Immunosensor for Detection of Prostate Specific Antigen based on Aminated Graphene Quantum Dots and Carboxyl Graphene Quantum Dots.

    Science.gov (United States)

    Wu, Dan; Liu, Yixin; Wang, Yaoguang; Hu, Lihua; Ma, Hongmin; Wang, Guoqin; Wei, Qin

    2016-02-04

    Prostate-specific antigen (PSA) was used as the model, an ultrasensitive label-free electrochemiluminescent immunosensor was developed based on graphene quantum dots. Au/Ag-rGO was sythsized and used as electrode material to load a great deal of graphene quantum dots due to the large surface area and excellent electron conductivity. After aminated graphene quantum dots and acarboxyl graphene quantum dots were modified onto the electrode, the ECL intensity was much high using K2S2O8 as coreactant. Then, antibody of PSA was immobilized on the surface of modified electrode surface through the adsorption of Au/Ag toward proteins, leading to the decrease of the ECL intensity. As proven by ECL spectra test and electrochemical impedance spectroscopy (EIS) analysis, the fabrication process of the immunosensor is successful. Under the optimal conditions, the ECL intensity decreased linearly with the logarithm of PSA concentration in the range of 1 pg/mL ~ 10 ng/mL. The detection limit achieved is 0.29 pg/mL. The immunosensor results were validated through the detection of PSA in serum samples with satisfactory results. Due to excellent stability, high sensitivity, acceptable repeatability and selectivity, the immunosensor has promising applications in disease and drug analysis.

  2. Sandwich-type electrochemical immunosensor for the detection of AFP based on Pd octahedral and APTES-M-CeO₂-GS as signal labels.

    Science.gov (United States)

    Wei, Yicheng; Li, Yan; Li, Na; Zhang, Yong; Yan, Tao; Ma, Hongmin; Wei, Qin

    2016-05-15

    In the present work, an ultrasensitive sandwich-type electrochemical immunosensor based on a novel signal amplification strategy was designed for quantitative detection of alpha fetoprotein (AFP). Au nanoparticles with biocompatibility were electrodeposited on the surface of glassy carbon electrode (GCE) which can effectively capture and immobilize primary anti-AFP (Ab1) to significantly amplify the electrochemical signal. Graphene Oxide and CeO2 mesoporous nanocomposite functionalized by the 3-aminopropyltriethoxysilane supported Pd octahedral nanoparticles (Pd/APTES-M-CeO2-GS) were utilized as labels of detection anti-AFP (Ab2). Pd octahedral nanoparticles presented good catalytic activity towards the reduction of H2O2. Due to the large specific surface area and good adsorption properties of APTES-CeO2-GS nanocomposite, large amount of Pd octahedral nanoparticles could be immobilized, which could amplify the electrochemical signal and improve the sensitivity of the immunosensor. Under optimal conditions, the immunosensor exhibited wide linear range from 0.1 pg/mL to 50 ng/mL with a low detection limit of 0.033 pg/mL (S/N=3) for AFP detection. In addition, high sensitivity, excellent selectivity, good reproducibility and stability were obtained for the immunosensor, which has a promising application for quantitative detection of other tumor markers in clinical diagnosis.

  3. Amperometric Biosensors Based on Carbon Paste Electrodes Modified with Nanostructured Mixed-valence Manganese Oxides and Glucose Oxidase

    Energy Technology Data Exchange (ETDEWEB)

    Cui, Xiaoli; Liu, Guodong; Lin, Yuehe

    2005-06-01

    Nanostructured multivalent manganese oxides octahedral molecular sieve (OMS), including cryptomelane-type manganese oxides and todorokite-type manganese oxides, were synthesized and evaluated for chemical sensing and biosensing at low operating potential. Both cryptomelane-type manganese oxides and todorokite-type manganese oxides are nanofibrous crystals with sub-nanometer open tunnels that provide a unique property for sensing applications. The electrochemical and electrocatalytic performance of OMS for the oxidation of H2O2 have been compared. Both cryptomelane-type manganese oxides and todorokite-type manganese oxides can be used to fabricate sensitive H2O2 sensors. Amperometric glucose biosensors are constructed by bulk modification of carbon paste electrodes (CPEs) with glucose oxidase as a biocomponent and nanostructured OMS as a mediator. A Nafion thin film was applied as an immobilization/encapsulation and protective layer. The biosensors were evaluated as an amperometric glucose detector at phosphate buffer solution with a pH 7.4 at an operating potential of 0.3 V (vs. Ag/AgCl). The biosensor is characterized by a well-reproducible amperometric response, linear signal-to-glucose concentration range up to 3.5 mM and 1.75 mM, and detection limits (S/N = 3) of 0.1 mM and 0.05 mM for todorokite-type manganese oxide and cryptomelane-type manganese oxide modified electrodes, respectively. The biosensors based on OMS exhibit considerable good reproducibility and stability, and the construction and renewal are simple and inexpensive.

  4. Amperometric biosensors based on carbon paste electrodes modified with nanostructured mixed-valence manganese oxides and glucose oxidase.

    Science.gov (United States)

    Cui, Xiaoli; Liu, Guodong; Lin, Yuehe

    2005-06-01

    Nanostructured, multivalent, manganese-oxide octahedral molecular sieves (OMS), including cryptomelane-type manganese oxides and todorokite-type manganese oxides, were synthesized and evaluated for chemical sensing and biosensing at low operating potential. Both cryptomelane-type manganese oxides and todorokite-type manganese oxides are nanofibrous crystals with subnanometer open tunnels that provide a unique property for sensing applications. The electrochemical and electrocatalytic performance of OMS for the oxidation of H2O2 have been compared. Both cryptomelane-type manganese oxides and todorokite-type manganese oxides can be used to fabricate sensitive H2O2 sensors. With glucose oxidase (GOx) as an enzyme model, amperometric glucose biosensors are constructed by bulk modification of carbon paste electrodes with GOx as a biocomponent and nanostructured OMS as a mediator. A Nafion thin film was applied as an immobilization/encapsulation and protective layer. The biosensors were evaluated as an amperometric glucose detector at phosphate buffer solution with a pH 7.4 at an operating potential of 0.3 V (vs Ag/AgCl). The biosensor is characterized by a well-reproducible amperometric response, linear signal-to-glucose concentration range up to 3.5 mmol/L and 1.75 mmol/L, and detection limits (S/N = 3) of 0.1 mmol/L and 0.05 mmol/L for todorokite-type manganese oxide and cryptomelane-type manganese oxide-modified electrodes, respectively. The biosensors based on OMS exhibit considerable good reproducibility and stability, and the construction and renewal are simple and inexpensive.

  5. Label-free C-reactive protein electronic detection with an electrolyte-gated organic field-effect transistor-based immunosensor.

    Science.gov (United States)

    Magliulo, Maria; De Tullio, Donato; Vikholm-Lundin, Inger; Albers, Willem M; Munter, Tony; Manoli, Kyriaki; Palazzo, Gerardo; Torsi, Luisa

    2016-06-01

    In this contribution, we propose a label-free immunosensor, based on a novel type of electrolyte-gated field-effect transistor (EGOFET), for ultrasensitive detection of the C-reactive protein (CRP). The recognition layer of the biosensor is fabricated by physical adsorption of the anti-CRP monoclonal antibody onto a poly-3-hexyl thiophene (P3HT) organic semiconductor surface. A supplementary nonionic hydrophilic polymer is used as a blocking agent preventing nonspecific interactions and allowing a better orientation of the antibodies immobilized onto the P3HT surface. The whole biomolecule immobilization procedure does not require any pretreatment of the organic semiconductor surface, and the whole functionalization process is completed in less than 30 min. Surface plasmon resonance (SPR) measurements were performed to assess the amount of biomolecules physisorbed onto the P3HT and to evaluate the CRP binding proprieties of the deposited anti-CRP layer. A partial surface coverage of about 23 % of adsorbed antibody molecules was found to most efficiently sense the CRP. The electrical performance of the EGOFET immunosensor was comparable to that of a bare P3HT EGOFET device, and the obtained CRP calibration curve was linear over six orders of magnitude (from 4 pM to 2 μM). The relative standard deviation of the individual calibration points, measured on immunosensors fabricated on different chips, ranged between 1 and 14 %, and a detection limit of 2 pM (220 ng/L) was established. The novel electronic immunosensor is compatible with low-cost fabrication procedures and was successfully employed for the detection of the CRP biomarker in the clinically relevant matrix serum. Graphical abstract Schematic of the EGOFET immunosensor for CRP detection. The anti-CRP monoclonal antibody layer is physisorbed on the P3HT organic semiconductor and the CRP is directly measured by a label-free electronic EGOFET transducer.

  6. A Label-Free Immunosensor for Ultrasensitive Detection of Ketamine Based on Quartz Crystal Microbalance

    Directory of Open Access Journals (Sweden)

    Ya Yang

    2015-04-01

    Full Text Available In this study, we have developed a label-free immunosensor with the variation of resonance frequency (Δf of a quartz crystal microbalance (QCM as readout signal for ultrasensitive detection of Ketamine (KT. An optimized strategy for immobilization of KT antibody on the surface of the QCM chip was implemented via the self-assembly modification of 3-mercaptopropionic acid, and then activated with 1-ethyl-3- (3-dimethylaminoprophl carbodiimide and n-hydroxysuccinimide. The specific affinity between the antibody and the antigen ensured a selective response toward KT. The Δf linearly related to the concentration of KT in the range of 1 to 40 pg/mL, with a detection limit of 0.86 pg/mL (S/N = 3. The obtained immunosensor was applied to detect the KT in spiked human urine without any pretreatment but dilution with recoveries from 91.8% to 108%. The developed sensor is promising to perform the portable or on-spot KT detection in clinic or forensic cases.

  7. Functionalised Au Coated Iron Oxide Nanocomposites Based Reusable Immunosensor for AFB1 Detection

    Directory of Open Access Journals (Sweden)

    Ruchika Chauhan

    2015-01-01

    Full Text Available A reusable sandwiched electrochemical piezoelectric immunosensor has been developed for aflatoxin B1 (AFB1 detection using gold coated iron oxide core-shell (Au-Fe3O4 nanostructure. The monoclonal anti-aflatoxin antibody (aAFB1 was immobilized on self-assembled monolayer of 4-aminothiophenol on gold coated quartz crystal to fabricate immunoelectrode (BSA/aAFB1/4-ATP/Au. In addition, secondary rabbit-immunoglobulin antibodies (r-IgGs functionalized with Au-Fe3O4 NPs via cysteamine (r-IgG-Cys-Au-Fe3O4 were allowed to interact with AFB1. Both competitive and noncompetitive strategies were employed and a competition between coated AFB1 and free AFB1 was carried out. The competitive mode shows higher linear range (0.05 to 5 ng mL−1 than the noncompetitive one (0.5 to 5 ng mL−1, high sensitivity 335.7 µA ng−1 mL cm−2, and LOD 0.07 ng mL−1. The fabricated immunosensor has been tested using cereal samples spiked with different concentrations of AFB1. The developed competitive immunoelectrode displays good reproducibility, and storage stability and regenerated with negligible loss in activity through removal of the r-IgG-Cys-Au-Fe3O4 conjugate using a strong external magnet.

  8. Effective antibodies immobilization and functionalized nanoparticles in a quartz-crystal microbalance-based immunosensor for the detection of parathion

    Science.gov (United States)

    Della Ventura, Bartolomeo; Iannaccone, Marco; Funari, Riccardo; Pica Ciamarra, Massimo; Altucci, Carlo; Capparelli, Rosanna; Roperto, Sante; Velotta, Raffaele

    2017-01-01

    Background Biosensor-based detection provides a rapid and low-cost alternative to conventional analytical methods for revealing the presence of the contaminants in water as well as solid matrices. Although important to be detected, small analytes (few hundreds of Daltons) are an issue in biosensing since the signal they induce in the transducer, and specifically in a Quartz-Crystal Microbalance, is undetectable. A pesticide like parathion (M = 292 Da) is a typical example of contaminant for which a signal amplification procedure is desirable. Methods/Findings The ballasting of the analyte by gold nanoparticles has been already applied to heavy target as proteins or bacteria to improve the limit of detection. In this paper, we extend the application of such a method to small analytes by showing that once the working surface of a Quartz-Crystal Microbalance (QCM) has been properly functionalized, a limit of detection lower than 1 ppb is reached for parathion. The effective surface functionalization is achieved by immobilizing antibodies upright oriented on the QCM gold surface by a simple photochemical technique (Photonic Immobilization Technique, PIT) based on the UV irradiation of the antibodies, whereas a simple protocol provided by the manufacturer is applied to functionalize the gold nanoparticles. Thus, in a non-competitive approach, the small analyte is made detectable by weighing it down through a “sandwich protocol” with a second antibody tethered to heavy gold nanoparticles. The immunosensor has been proved to be effective against the parathion while showing no cross reaction when a mixture of compounds very similar to parathion is analyzed. Conclusion/Significance The immunosensor described in this paper can be easily applied to any small molecule for which polyclonal antibodies are available since both the functionalization procedure of the QCM probe surface and gold nanoparticle can be applied to any IgG, thereby making our device of general

  9. Highly sensitive amperometric biosensor based on electrochemically-reduced graphene oxide-chitosan/hemoglobin nanocomposite for nitromethane determination.

    Science.gov (United States)

    Wen, Yunping; Wen, Wei; Zhang, Xiuhua; Wang, Shengfu

    2016-05-15

    Nitromethane (CH3NO2) is an important organic chemical raw material with a wide variety of applications as well as one of the most common pollutants. Therefore it is pretty important to establish a simple and sensitive detection method for CH3NO2. In our study, a novel amperometric biosensor for nitromethane (CH3NO2) based on immobilization of electrochemically-reduced graphene oxide (rGO), chitosan (CS) and hemoglobin (Hb) on a glassy carbon electrode (GCE) was constructed. Scanning electron microscopy, infrared spectroscopy and electrochemical methods were used to characterize the Hb-CS/rGO-CS composite film. The effects of scan rate and pH of phosphate buffer on the biosensor have been studied in detail and optimized. Due to the graphene and chitosan nanocomposite, the developed biosensor demonstrating direct electrochemistry with faster electron-transfer rate (6.48s(-1)) and excellent catalytic activity towards CH3NO2. Under optimal conditions, the proposed biosensor exhibited fast amperometric response (biosensor had high selectivity, reproducibility and stability, providing the possibility for monitoring CH3NO2 in complex real samples.

  10. SPR investigations of the formation of intermediate layer of the immunosensor bioselective element based on the recombinant Staphylococcal protein A

    Directory of Open Access Journals (Sweden)

    Rachkov A. E.

    2015-08-01

    Full Text Available Aim. To investigate the formation of an intermediate layer of the immunosensor bioselective element based on the recombinant protein A from Staphylococcus aureus with cysteine residue (SPA-Cys and its interactions with human IgG using the SPR spectrometer «Plasmon». Methods. The activity of the immune components applied was tested by ELISA. The spectrometry of surface plasmon resonance was used for studying protein immobilization on a gold sensor surface and interactions between the immobilized SPA-Cys and human immunoglobulin. Results. A direct dependence of the sensor response on the concentration of SPA-Cys in the range of 0.2 to 2 µM at its immobilization was demonstrated. The efficiency of blocking nonspecific adsorption sites on the sensor surface with milk proteins and the direct dependence of the sensor response on IgG concentration and surface density of immobilized SPA-Cys were shown. Fitting the experimental data to a Langmuir plot yields a Kd value for SPA-Cys/IgG binding 8.5 ± 0.7× 10-8 M (Ka = 1.2 ± 0.1× 107 M–1. The determined equilibrium binding constant indicates a quite strong interaction and its value is consistent with the literature data. Conclusions. A successful immobilization of SPA-Cys on a gold surface of the SPR spectrometer while preserving its high immunoglobulin-binding activity, selectivity and stability of the sensor response confirms the efficiency of SPA-Cys as an intermediate component for the creation of the immunosensor bioselective elements.

  11. Electrochemical immunosensor for ethinylestradiol using diazonium salt grafting onto silver nanoparticles-silica-graphene oxide hybrids.

    Science.gov (United States)

    Cincotto, Fernando H; Martínez-García, Gonzalo; Yáñez-Sedeño, Paloma; Canevari, Thiago C; Machado, S A S; Pingarrón, José M

    2016-01-15

    This work describes the preparation of an electrochemical immunosensor for ethinylestradiol (EE2) based on grafting of diazonium salt of 4-aminobenzoic acid onto a glassy carbon electrode modified with silver nanoparticles/SiO2/graphene oxide hybrid followed by covalent binding of anti-ethinylestradiol (anti-EE2) to activated carboxyl groups. A competitive immunoassay was developed for the determination of the hormone using peroxidase-labeled ethinylestradiol (HRP-EE2) and measurement of the amperometric response at -200mV in the presence of hydroquinone (HQ) as redox mediator. The calibration curve for EE2 exhibited a linear range between 0.1 and 50ng/mL (r(2)=0.996), with a detection limit of 65pg/mL. Interference studies with other hormones related with EE2 revealed the practical specificity of the developed method for the analyte. A good reproducibility, with RSD=4.5% (n=10) was also observed. The operating stability of a single bioelectrode modified with anti-EE2 was maintained at least for 15 days when it was stored at 4°C under humid conditions between measurements. The developed immunosensor was applied to the analysis of spiked urine with good results.

  12. Rapid amperometric detection of trace metals by inhibition of an ultrathin polypyrrole-based glucose biosensor.

    Science.gov (United States)

    Ayenimo, Joseph G; Adeloju, Samuel B

    2016-02-01

    A sensitive and reliable inhibitive amperometric glucose biosensor is described for rapid trace metal determination. The biosensor utilises a conductive ultrathin (55 nm thick) polypyrrole (PPy) film for entrapment of glucose oxidase (GOx) to permit rapid inhibition of GOx activity in the ultrathin film upon exposure to trace metals, resulting in reduced glucose amperometric response. The biosensor demonstrates a relatively fast response time of 20s and does not require incubation. Furthermore, a complete recovery of GOx activity in the ultrathin PPy-GOx biosensor is quickly achieved by washing in 2mM EDTA for only 10s. The minimum detectable concentrations achieved with the biosensor for Hg(2+), Cu(2+), Pb(2+) and Cd(2+) by inhibitive amperometric detection are 0.48, 1.5, 1.6 and 4.0 µM, respectively. Also, suitable linear concentration ranges were achieved from 0.48-3.3 µM for Hg(2+), 1.5-10 µM for Cu(2+), 1.6-7.7 µM for Pb(2+) and 4-26 µM for Cd(2+). The use of Dixon and Cornish-Bowden plots revealed that the suppressive effects observed with Hg(2+) and Cu(2+) were via non-competitive inhibition, while those of Pb(2+) and Cd(2+) were due to mixed and competitive inhibition. The stronger inhibition exhibited by the trace metals on GOx activity in the ultrathin PPy-GOx film was also confirmed by the low inhibition constant obtained from this analysis. The biosensor was successfully applied to the determination of trace metals in tap water samples.

  13. A Novel Amperometric Nitric Oxide Sensor Based on Polythionine /Nation Modified Glassy Carbon Electrode

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    A novel amperometric sensor for the determination of nitric oxide was developed by coating polythionine / nafion on a glassy carbon electrode. This sensor exhibited a great enhancement to the oxidation of nitric oxide. The oxidation peak currents were linear to the concentration of nitric oxide over the wide range from 3.6×10-7 to 6.8×10-5 mol. L-1, and the detection limit was 7.2×10-8 mol. L-1. Experimental results showed that this nitric oxide sensor possessed excellent selectivity and longer stability. NO releasing from rat kidney was monitored by this sensor.

  14. Study on Rhizoma Chuanxiong based on capillary electrophoresis with amperometric detection

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    A high-performance capillary electrophoresis with amperometric detection(CE-AD) method has been developed for the analysis of seven bioactive ingredients,namely ferulic acid(FA),vanillin,vanillic acid,p-hydroxybenzoic acid,caffeic acid,gallic acid and protocatechuic acid,in Rhizoma Chuanxiong.The effects of several factors such as the acidity and concentration of running buffer,the separation voltage,the applied potential to working electrode and the injection time were investigated.Under the optimum con...

  15. Immunosensor based on nanocomposite of nanostructured zirconium oxide and gelatin-A.

    Science.gov (United States)

    Bagbi, Yana; Sharma, Anshu; Bohidar, H B; Solanki, Pratima R

    2016-01-01

    We have reported the studies related to the fabrication of a nanocomposite, comprising of sol-gel derived inorganic zirconium oxide nanoparticles (ZrO2 NPs) and organic biopolymer gelatin-A (GA), deposited on indium-tin-oxide (ITO) coated glass substrate by drop casting method. The GA-ZrO2/ITO electrode was used for immobilization of monoclonal antibodies (Ab) specific to antigen Vibrio cholerae (Vc) followed by bovine serum albumin (BSA) for antigen Vc detection using electrochemical techniques. The structural and morphological behaviour of these ZrO2 NPs, GA-ZrO2/ITO electrode and BSA/Ab/GA-ZrO2/ITO immunosensor was characterized by scanning electron microscopy and Fourier transform infrared spectroscopy techniques. The transmission electron microscopy study exhibited a spherical shape ZrO2 NPs. The average crystalline size of ZrO2 NPs was obtained as 10.3 ± 1 nm from X-ray diffraction measurement and 72 nm hydrodynamic radius measured by dynamic light scattering. GA-ZrO2 nanocomposite provides a porous structure which assists to higher loading of Ab on the matrix surface that improved the biosensing properties. The electrochemical response studies of the fabricated BSA/Ab/GA-ZrO2/ITO immunosensor exhibited good linearity in the range of 50-400 ng mL(-1), low limit of detection of 0.74 ng/mL, sensitivity as 0.03 Ω ng(-1)mL(-1)cm(-2) and reproducibility more than 10 times.

  16. Miniaturized flow system based on enzyme modified PMMA microreactor for amperometric determination of glucose.

    Science.gov (United States)

    Cerdeira Ferreira, Luís Marcos; da Costa, Eric Tavares; do Lago, Claudimir Lucio; Angnes, Lúcio

    2013-09-15

    This paper describes the development of a microfluidic system having as main component an enzymatic reactor constituted by a microchannel assembled in poly(methyl methacrylate) (PMMA) substrate connected to an amperometric detector. A CO2 laser engraving machine was used to make the channels, which in sequence were thermally sealed. The internal surfaces of the microchannels were chemically modified with polyethyleneimine (PEI), which showed good effectiveness for the immobilization of the glucose oxidase enzyme using glutaraldehyde as crosslinking agent, producing a very effective microreactor for the detection of glucose. The hydrogen peroxide generated by the enzymatic reaction was detected in an electrochemical flow cell localized outside of the reactor using a platinum disk as the working electrode. The proposed system was applied to the differential amperometric determination of glucose content in soft drinks showing good repeatability (DPR=1.72%, n=50), low detection limit (1.40×10(-6)molL(-1)), high sampling frequency (calculated as 345 samples h(-1)), and relatively good stability for long-term use. The results were in close agreement with those obtained by the classical spectrophotometric method utilized to quantify glucose in biological fluids.

  17. Development of Amperometric Biosensors Based on Nanostructured Tyrosinase-Conducting Polymer Composite Electrodes

    Directory of Open Access Journals (Sweden)

    Francisco Javier del Campo

    2013-05-01

    Full Text Available Bio-composite coatings consisting of poly(3,4-ethylenedioxythiophene (PEDOT and tyrosinase (Ty were successfully electrodeposited on conventional size gold (Au disk electrodes and microelectrode arrays using sinusoidal voltages. Electrochemical polymerization of the corresponding monomer was carried out in the presence of various Ty amounts in aqueous buffered solutions. The bio-composite coatings prepared using sinusoidal voltages and potentiostatic electrodeposition methods were compared in terms of morphology, electrochemical properties, and biocatalytic activity towards various analytes. The amperometric biosensors were tested in dopamine (DA and catechol (CT electroanalysis in aqueous buffered solutions. The analytical performance of the developed biosensors was investigated in terms of linear response range, detection limit, sensitivity, and repeatability. A semi-quantitative multi-analyte procedure for simultaneous determination of DA and CT was developed. The amperometric biosensor prepared using sinusoidal voltages showed much better analytical performance. The Au disk biosensor obtained by 50 mV alternating voltage amplitude displayed a linear response for DA concentrations ranging from 10 to 300 μM, with a detection limit of 4.18 μM.

  18. Non-invasive determination of glucose directly in raw fruits using a continuous flow system based on microdialysis sampling and amperometric detection at an integrated enzymatic biosensor.

    Science.gov (United States)

    Vargas, E; Ruiz, M A; Campuzano, S; Reviejo, A J; Pingarrón, J M

    2016-03-31

    A non-destructive, rapid and simple to use sensing method for direct determination of glucose in non-processed fruits is described. The strategy involved on-line microdialysis sampling coupled with a continuous flow system with amperometric detection at an enzymatic biosensor. Apart from direct determination of glucose in fruit juices and blended fruits, this work describes for the first time the successful application of an enzymatic biosensor-based electrochemical approach to the non-invasive determination of glucose in raw fruits. The methodology correlates, through previous calibration set-up, the amperometric signal generated from glucose in non-processed fruits with its content in % (w/w). The comparison of the obtained results using the proposed approach in different fruits with those provided by other method involving the same commercial biosensor as amperometric detector in stirred solutions pointed out that there were no significant differences. Moreover, in comparison with other available methodologies, this microdialysis-coupled continuous flow system amperometric biosensor-based procedure features straightforward sample preparation, low cost, reduced assay time (sampling rate of 7 h(-1)) and ease of automation.

  19. A novel amperometric catechol biosensor based on α-Fe2O3 nanocrystals-modified carbon paste electrode.

    Science.gov (United States)

    Sarika, C; Shivakumar, M S; Shivakumara, C; Krishnamurthy, G; Narasimha Murthy, B; Lekshmi, I C

    2017-05-01

    In this work, we designed an amperometric catechol biosensor based on α-Fe2O3 nanocrystals (NCs) incorporated carbon-paste electrode. Laccase enzyme is then assembled onto the modified electrode surface to form a nanobiocomposite enhancing the electron transfer reactions at the enzyme's active metal centers for catechol oxidation. The biosensor gave good sensitivity with a linear detection response in the range of 8-800 μM with limit of detection 4.28 μM. We successfully employed the sensor for real water sample analysis. The results illustrate that the metal oxide NCs have enormous potential in the construction of biosensors for sensitive determination of phenol derivatives.

  20. Determination of Patulin Using Amperometric Tyrosinase Biosensors Based on Electrodes Modified with Carbon Nanotubes and Gold Nanoparticles

    Directory of Open Access Journals (Sweden)

    R.M. Varlamova

    2016-06-01

    Full Text Available New amperometric biosensors based on platinum screen printed electrodes modified with multi-walled carbon nanotubes, gold nanoparticles, and immobilized enzyme – tyrosinase have been developed for determination of patulin in the concentrations of 1·10–6 – 8·10–12 mol/L with an error of no more than 0.063. The best conditions for obtaining gold nanoparticles have been chosen. The conditions for immobilization of multi-walled carbon nanotubes and gold nanoparticles on the surface of the planar electrode have been revealed. The conditions for functioning of the proposed biosensors have been identified. The results have been used to control the content of patulin in food products within and lower than the maximum allowable levels.

  1. Rapid detection of Escherichia coli O157:H7 and Salmonella Typhimurium in foods using an electrochemical immunosensor based on screen-printed interdigitated microelectrode and immunomagnetic separation.

    Science.gov (United States)

    Xu, Meng; Wang, Ronghui; Li, Yanbin

    2016-01-01

    Foodborne pathogens have continuously been a serious food safety issue and there is a growing demand for a rapid and sensitive method to screen the pathogens for on-line or in-field applications. Therefore, an impedimetric immunosensor based on the use of magnetic beads (MBs) for separation and a screen-printed interdigitated microelectrode (SP-IDME) for measurement was studied for the rapid detection of Escherichia coli O157:H7 and Salmonella Typhimurium in foods. Streptavidin coated MBs were functionalized with corresponding biotinylated antibodies (Ab) to capture the target bacteria. The glucose oxidase (GOx)-Ab conjugates were employed to label the MBs-Ab-cell complexes. The yielded MBs-Ab-cell-Ab-GOx biomass was mixed with the glucose solution to trigger an enzymatic reaction which produced gluconic acid. This increased the ion strength of the solution, thus decreasing the impedance of the solution measured on the SP-IDME. Our results showed that the immunosensor was capable of specifically detecting E. coli O157:H7 and S. Typhimurium within the range of 10(2)-10(6) cfu ml(-1) in the pure culture samples. E. coli O157:H7 in ground beef and S. Typhimurium in chicken rinse water were also examined. The limits of detection (LODs) for the two bacteria in foods were 2.05×10(3) cfu g(-1) and 1.04×10(3) cfu ml(-1), respectively. This immunosensor required only a bare electrode to measure the impedance changes, and no surficial modification on the electrode was needed. It was low-cost, reproducible, easy-to-operate, and easy-to-preserve. All these merits demonstrated this immunosensor has great potential for the rapid and on-site detection of pathogenic bacteria in foods.

  2. Sensitive detection of microcystin-LR by using a label-free electrochemical immunosensor based on Au nanoparticles/silicon template/methylene blue nanocomposite.

    Science.gov (United States)

    Fu, Xuewen; Feng, Yajuan; Niu, Sipeng; Zhao, Chunling; Yang, Minghui; Yang, Yunhui

    2013-12-01

    A label-free electrochemical immunosensor, based on a gold nanoparticles (Au NPs)/silicon template/methylene blue (MB)/chitosan (CHIT) nanocomposite-modified electrode, was fabricated for the ultrasensitive detection of microcystin-LR (MC-LR). The nanocomposite film showed high binding affinity to the antibodies of MC-LR because gold nanoparticles have large surface area and good biocompatibility which can immobilize large amount of antibody through ionic interactions and other interactions between AuNPs and mercapto or primary amine groups of antibodies with high stability and bioactivity. MB was used as redox indicator due to its good electrochemical behavior in conductive substrate. This hybrid membrane was evaluated by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and differential pulse voltammetry (DPV) to determine its electrochemical properties in immunosensor application. A decrease in DPV responses was observed with increasing concentrations of MC-LR in standard and real samples due to the formation of immuno-complexes between MC-LR and anti-MC-LR antibodies which hindered the electron charge transfer at the electrode-electrolyte interface. At optimal conditions, this immunosensor could detect MC-LR in a linear range from 0.5 ng/mL to 25 microg/mL with a low detection limit of 0.1 ng/mL at 3 sigma. Moreover, the prepared immunosensor was applied for the analysis of MC-LR in water samples with satisfactory results. The proposed method showed high selectivity, acceptable reproducibility, stability and reliability.

  3. Immunosensors in Clinical Laboratory Diagnostics.

    Science.gov (United States)

    Justino, Celine I L; Duarte, Armando C; Rocha-Santos, Teresa A P

    2016-01-01

    The application of simple, cost-effective, rapid, and accurate diagnostic technologies for detection and identification of cardiac and cancer biomarkers has been a central point in the clinical area. Biosensors have been recognized as efficient alternatives for the diagnostics of various diseases due to their specificity and potential for application on real samples. The role of nanotechnology in the construction of immunological biosensors, that is, immunosensors, has contributed to the improvement of sensitivity, since they are based in the affinity between antibody and antigen. Other analytes than biomarkers such as hormones, pathogenic bacteria, and virus have also been detected by immunosensors for clinical point-of-care applications. In this chapter, we first introduced the various types of immunosensors and discussed their applications in clinical diagnostics over the recent 6 years, mainly as point-of-care technologies for the determination of cardiac and cancer biomarkers, hormones, pathogenic bacteria, and virus. The future perspectives of these devices in the field of clinical diagnostics are also evaluated.

  4. Amperometric Determination of Indole-3-acetic Acid Based on Platinum Nanowires and Carbon Nanotubes

    Institute of Scientific and Technical Information of China (English)

    Ruo Zhong WANG; Lang Tao XIAO; Ming Hui YANG; Jun Hui DING; Feng Li QU; Guo Li SHEN

    2006-01-01

    Platinum nanowire (PtNW) can be grown by electrodeposition in polycarbonate membrane, with the average diameter of the nanowires about 250 nm. The PtNW and multiwalled carbon nanotubes (CNT) are then dispersed into chitosan (CHIT) solution. The resulting PtNW-CNT-CHIT material brings new capabilities for electrochemical devices by using the synergistic action of the electrocatalytic activity of PtNW and CNT. By dropping the PtNW-CNT-CHIT film onto the glassy carbon (GC) electrode surface, and after evaporationan amperometric sensor for the determination of indole-3-acetic acid (IAA) was developed. The oxidation current of IAA increased significantly at the PtNW-CNT-CHIT film coated GC electrode,in contrast to that at the CNT-CHIT modified GC. The linear response of the sensor is from 50ng/ml to 50 μg/ml with a detection limit of 25 ng/mL.

  5. An electrochemical immunosensor for quantitative detection of ficolin-3

    Science.gov (United States)

    San, Lili; Zeng, Dongdong; Song, Shiping; Zuo, Xiaolei; Zhang, Huan; Wang, Chenguang; Wu, Jiarui; Mi, Xianqiang

    2016-06-01

    Diabetes mellitus (DM) is one of the most common metabolic disorders in the world, of which more than 90% is type-2 diabetes mellitus (T2DM). There is a rather urgent need for reliable, sensitive and quick detection techniques in clinical application of T2DM. Ficolin-3 is a potential biomarker of T2DM, because serum ficolin-3 levels are associated with insulin resistance and predict the incidence of T2DM. Herein, a sandwich-type electrochemical immunosensor was developed for the detection of ficolin-3 in human serum. Cyclic voltammetry and the amperometric current versus time were used to characterize the performance of the immunosensor. Under optimal conditions, the detection limitation of ficolin-3 was 100 ng ml-1 and the linear dynamic range was between 2 and 50 μg ml-1. The method has ideal accuracy, excellent stability and selectivity and has wide application prospects in clinical research.

  6. Single-Multiplex Detection of Organ Injury Biomarkers using SPRi based Nano-Immunosensor

    Science.gov (United States)

    Zeidan, Effat; Li, Siqi; Zhou, Zhiguo; Miller, Jennifer; Sandros, Marinella G.

    2016-01-01

    The clinical assessment of multiple organ dysfunctions at early stages is recognized to be an important factor in prompting definitive treatment decisions that prevent irreversible organ damage. In this article, we propose a real-time, label-free, and multiplex nanoenhanced SPRi platform to quantitatively assess two biomarkers, kidney injury molecule (KIM-1) and high mobility group box-1 (HMGB-1) simultaneously in buffer. Our work involves three major contributions in the design of the immunosensor: (1) we applied site-specific immobilization of antibodies to the solid surface that avoids loss of biological activity caused by covalent attachment; (2) we constructed a well-blocked sensor surface that exhibits minimal non-specific adsorption for singleplex measurements of each biomarker in buffer; and (3) we adopted a sandwich assay that implements functionalized quantum dots (NanoEnhancers) as signal amplifiers to achieve a sensitivity level of 5 pg/mL for KIM-1 and HMGB-1 in buffer. We foresee great potential and success in extending this multiplex and ultra-sensitive platform to assess a variety of other emerging clinical biomarkers at low concentrations and in complex matrices. PMID:27796342

  7. Diagnosis of schistosomiasis japonica with interfacial co-assembly-based multi-channel electrochemical immunosensor arrays.

    Science.gov (United States)

    Deng, Wangping; Xu, Bin; Hu, Haiyan; Li, Jianyong; Hu, Wei; Song, Shiping; Feng, Zheng; Fan, Chunhai

    2013-01-01

    Schistosomiasis control remains to be an important and challenging task in the world. However, lack of quick, simple, sensitive and specific sero-diagnostic test is still a hurdle in the control practice. The commonly employed enzyme-linked immuno-sorbent assay (ELISA) relies on the native soluble egg antigen (SEA) that is limited in supply. Here we developed an electrochemical immunosensor array (ECISA) assay with an interfacial co-assembly strategy. A recombinant Schistosoma japonicum (Sj) calcium-binding protein (SjE16) was used as a principal antigen, while the SEA as a minor, co-assembling agent, with a ratio of 8:1 (SjE16: SEA, Sj16EA), which was co-immobilized on a disposable 16-channel screen-printed carbon electrode array. A portable electrochemical detector was employed to detect antibodies in serum samples. The sensitivity of ECISA reached 100% with minimal cross-reactions. Therefore, we have demonstrated that this rapid, sensitive and specific ECISA technique has the potential to perform large-scale on-site screening of Sj infection.

  8. Enzyme-free electrochemical immunosensor based on methylene blue and the electro-oxidation of hydrazine on Pt nanoparticles.

    Science.gov (United States)

    Dutta, Gorachand; Nagarajan, Sureshbabu; Lapidus, Lisa J; Lillehoj, Peter B

    2017-06-15

    Enzyme-free electrochemical sensors enable rapid, high sensitivity measurements without the limitations associated with enzyme reporters. However, the performance of non-enzymatic electrochemical sensors tends to suffer from slow electrode kinetics and poor signal stability. We report a new enzyme-free electrochemical immunosensor based on a unique competitive detection scheme using methylene blue (MB), hydrazine and platinum nanoparticles (Pt NPs). This scheme is coupled with a robust immunosandwich format employing a MB-labelled detection antibody as a non-enzymatic reporter. In the presence of the target antigen, surface-immobilized MB consumes interfacial hydrazine thereby diminishing the electro-oxidation of hydrazine on Pt NPs. Thus, the concentration of the antigen is directly proportional to the reduction in the electrochemical signal. For proof-of-concept, this sensor was used to detect Plasmodium falciparum histidine-rich protein 2 (PfHRP2), an important malaria biomarker, in unadulterated human saliva samples. Chronocoulometric measurements showed that this platform exhibits pM-range sensitivity, high specificity and good reproducibility, making it well suited for many biosensing applications including noninvasive diagnostic testing.

  9. Silica nanoparticle-based microfluidic immunosensor with laser-induced fluorescence detection for the quantification of immunoreactive trypsin.

    Science.gov (United States)

    Seia, Marco A; Stege, Patricia W; Pereira, Sirley V; De Vito, Irma E; Raba, Julio; Messina, Germán A

    2014-10-15

    The purpose of this study was to develop a silica nanoparticle-based immunosensor with laser-induced fluorescence (LIF) as a detection system. The proposed device was applied to quantify the immunoreactive trypsin (IRT) in cystic fibrosis (CF) newborn screening. A new ultrasonic procedure was used to extract the IRT from blood spot samples collected on filter papers. After extraction, the IRT reacted immunologically with anti-IRT monoclonal antibodies immobilized on a microfluidic glass chip modified with 3-aminopropyl functionalized silica nanoparticles (APSN-APTES-modified glass chips). The bounded IRT was quantified by horseradish peroxidase (HRP)-conjugated anti-IRT antibody (anti-IRT-Ab) using 10-acetyl-3,7-dihydroxyphenoxazine (ADHP) as enzymatic mediator. The HRP catalyzed the oxidation of nonfluorescent ADHP to highly fluorescent resorufin, which was measured by LIF detector, using excitation lambda at 561nm and emission at 585nm. The detection limits (LODs) calculated for LIF detection and for a commercial enzyme-linked immunosorbent assay (ELISA) test kit were 0.87 and 4.2ngml(-1), respectively. The within- and between-assay variation coefficients for the LIF detection procedure were below 6.5%. The blood spot samples collected on filter papers were analyzed with the proposed method, and the results were compared with those of the reference ELISA method, demonstrating a potential usefulness for the clinical assessment of IRT during the early neonatal period.

  10. Ultrasensitive detection of E. coli O157:H7 with biofunctional magnetic bead concentration via nanoporous membrane based electrochemical immunosensor.

    Science.gov (United States)

    Chan, Ka Yiu; Ye, Wei Wei; Zhang, Yu; Xiao, Li Dan; Leung, Polly H M; Li, Yi; Yang, Mo

    2013-03-15

    In this paper, biofunctional magnetic beads were investigated for bacterial cells concentration in a nanoporous alumina membrane based immunosensor for ultra-sensitive detection of E. coli O157:H7. The specific antibody modified magnetic beads were used for concentration of E coli O157:H7 from samples in a small region to enhance sensitivity. The magnetic bead conjugated E. coli O157:H7 cells were then captured on the nanoporous alumina membrane with immobilized antibody via assembled PEG-silane linker. Scanning electron microscopy and fluorescent microscopy were used to demonstrate the magnetic bead-bacteria cell conjugation and bacteria cells magnetic concentration, respectively. Impedance spectroscopy was used to monitor the pure E coli O157:H7 cells and magnetic bead conjugated E coli O157:H7 cells binding on antibody immobilized nanoporous membrane with or without magnetic field. Compared with direct detection of pure bacteria cells, this method via magnetic bead conjugation and concentration demonstrated the ultrasensitivity of 10 CFU/mL for E coli O157:H7 detection.

  11. New immunosensor for Lactoferrin determination in human milk and several pharmaceutical dairy milk products recommended for the unweaned diet.

    Science.gov (United States)

    Campanella, Luigi; Martini, Elisabetta; Tomassetti, Mauro

    2008-09-29

    Thorough research was carried out on Lactoferrin immunosensor development. Furthermore, two different competitive procedures were used for Lactoferrin determination, in which either the antigen (Lactoferrin) or the antibody (anti-Lactoferrin) was, respectively, conjugated with horseradish peroxidase enzyme using a biotinylation process. The biotinylation of Lactoferrin and the subsequently used competition procedure for the immunosensor measurement were to get ready. Three different kinds of immunosensors were implemented, in all cases using the peroxidase enzyme as marker and hydrogen peroxide as substrate, but alternatively using as transducers one of the following sensors: (i) an amperometric electrode for H2O2, (ii) a Clark electrode and (iii) an iodide electrode. After optimizing the "competitive" measurement procedures and the transducer, the new Lactoferrin immunosensor was used for the determination of Lactoferrin content in human milk and in different types of dried milks or other dairy products, specifically produced and sold in chemist's shops to feed unweaned children in the first few months of life.

  12. Highly Sensitive and Selective Amperometric Sensor for Iodate Based on 9,10-Phenanthrenequinone Derived Graphene

    Institute of Scientific and Technical Information of China (English)

    WANG Hong-juan; CHENG Na-na; YANG Xiao-yang; LI Xiao-meng; ZHU Lian-de

    2013-01-01

    We reported on a new amperometric sensor for the sensitive and selective determination of iodate in table salt.The iodate sensor was constructed by the integration of a novel nanocomposite which was made from 9,10-phenanthrenequinone(PQ) and graphene(GP) with a glassy carbon electrode(GCE).The synthesized graphene and the nanocomposite were well characterized by X-ray diffraction(XRD),transmission electron microscopy(TEM),Fourier transform infrared(FTIR) spectroscopy and Raman spectroscopy.We fully studied the electrochemical behavior and kinetic characteristics of the PQ/GP nanocomposite at GCE.The PQ/GP electrode shows a good electrochemical catalytic activity towards the reduction of iodate,which makes itself a sensitive and selective electrochemical sensor for iodate.The iodate sensor displays a high sensitivity(1.04 μA·μmol.L-1),a low detection limit(1.0× 10-8mol/L),a rapid response(less than 2 s),and a broad linear range(from 5.0× 10-8 mol/L to 6.0× 10-3 mol/L).In addition,the sensor is interference free.The practical application of the proposed sensor was tested by the detection of iodate in table salt.

  13. Development of an amperometric sulfite biosensor based on SO(x)/PBNPs/PPY modified ITO electrode.

    Science.gov (United States)

    Rawal, Rachna; Pundir, C S

    2012-11-01

    A sulfite oxidase (SO(x)) (EC 1.8.3.1) purified from Syzygium cumini leaves was immobilized onto prussian blue nanoparticles/polypyrrole composite (PBNPs/PPY) electrodeposited onto the surface of indium tin oxide (ITO) electrode. An amperometric sulfite biosensor was fabricated using SO(x)/PBNPs/PPY/ITO electrode as working electrode, Ag/AgCl as standard and Pt wire as auxiliary electrode connected through a potentiostat. The working electrode was characterized by Fourier transform infrared (FTIR) spectroscopy, cyclic voltammetry (CV), scanning electron microscopy (SEM) and electrochemical impedance spectroscopy (EIS) before and after immobilization of SO(x). The biosensor showed optimum response within 2s, when operated at 20 mV s⁻¹ in 0.1M Tris-HCl buffer, pH 8.5 and at 35 °C. Linear range and minimum detection limit were 0.5-1000 μM and 0.12 μM (S/N=3) respectively. There was good correlation (r=0.99) between red wine samples sulfite value by standard DTNB method and the present method. The sensor was evaluated with 97% recovery of added sulfite in red wine samples and 2.2% and 4.3% within and between batch coefficients of variation respectively. The sensor was employed for determination of sulfite level in red and white wine samples. The enzyme electrode was used 200 times over a period of 3 months when stored at 4 °C.

  14. Nonenzymatic amperometric sensor for ascorbic acid based on hollow gold/ruthenium nanoshells.

    Science.gov (United States)

    Jo, Ara; Kang, Minkyung; Cha, Areum; Jang, Hye Su; Shim, Jun Ho; Lee, Nam-Suk; Kim, Myung Hwa; Lee, Youngmi; Lee, Chongmok

    2014-03-28

    We report a new nonenzymatic amperometric detection of ascorbic acid (AA) using a glassy carbon (GC) disk electrode modified with hollow gold/ruthenium (hAu-Ru) nanoshells, which exhibited decent sensing characteristics. The hAu-Ru nanoshells were prepared by the incorporation of Ru on hollow gold (hAu) nanoshells from Co nanoparticle templates, which enabled AA selectivity against glucose without aid of enzyme or membrane. The structure and electrocatalytic activities of the hAu-Ru catalysts were characterized by spectroscopic and electrochemical techniques. The hAu-Ru loaded on GC electrode (hAu-Ru/GC) showed sensitivity of 426 μA mM(-1) cm(-2) (normalized to the GC disk area) for the linear dynamic range of <5 μM to 2 mM AA at physiological pH. The response time and detection limit were 1.6 s and 2.2 μM, respectively. Furthermore, the hAu-Ru/GC electrode displayed remarkable selectivity for ascorbic acid over all potential biological interferents, including glucose, uric acid (UA), dopamine (DA), 4-acetamidophenol (AP), and nicotinamide adenine dinucleotide (NADH), which could be especially good for biological sensing.

  15. A novel electrochemiluminescent immunosensor based on the quenching effect of aminated graphene on nitrogen-doped carbon quantum dots

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, Jing; Han, Tongqian; Ma, Hongmin; Yan, Tao; Pang, Xuehui [Key Laboratory of Chemical Sensing & Analysis in Universities of Shandong, School of Chemistry and Chemical Engineering, University of Jinan, Jinan 250022 (China); Li, Yueyun [School of Chemical Engineering, Shandong University of Technology, Zibo 255049 (China); Wei, Qin, E-mail: sdjndxwq@163.com [Key Laboratory of Chemical Sensing & Analysis in Universities of Shandong, School of Chemistry and Chemical Engineering, University of Jinan, Jinan 250022 (China)

    2015-08-19

    Nitrogen-doped carbon quantum dots (N-CQDs) with an average diameter of 2 nm were synthesized by carbonization of diethylene triamine pentacetate acid (DTPA). The simple prepared N-CQDs showed excellent electrochemiluminescence (ECL) property and were used as luminophors to fabricate a sandwich-type ECL immunosensor. Aminated graphene (NH{sub 2}-G) was also synthesized and used as a label of secondary antibody. The labeled NH{sub 2}-G could effectively quench the ECL of N-CQDs modified on electrodes due to ECL resonance energy transfer (ERET). Immunological recognition which induced ECL quenching enabled the quantitative determination of biomarkers. Alpha fetoprotein (AFP) was selected as a model analyte to investigate the analytical performance of the proposed immunosensor. Under optimal conditions, a good linear relationship between ECL intensity and the logarithm of AFP concentration was obtained in the range of 0.01–100 ng mL{sup −1} with the detection limit of 3.3 pg mL{sup −1}. The proposed ECL immunosensor showed good stability, acceptable selectivity and reproducibility. - Highlights: • ECL behavior of N-CQDs was investigated. • NH{sub 2}-G for quenching N-CQDs emission was investigated. • The linearly range of the immunosensor for AFP was 0.01 ng/mL–100 ng/mL.

  16. A new method for non-labeling attomolar detection of diseases based on an individual gold nanorod immunosensor

    DEFF Research Database (Denmark)

    Phuoc Long, Truong; Cao, Cuong; Park, Sungho;

    2011-01-01

    the cetyltrimethylammonium bromide (CTAB), a tightly packed self-assembled monolayer of HS (CH2)11(OCH2CH2)6OCH2COOH(OEG6) has been successfully formed on the gold nanorod surface prior to the LSPR sensing, leading to the successful fabrication of individual gold nanorod immunosensors. Using prostate specific antigen (PSA...

  17. Amperometric L-glutamate biosensor based on bacterial cell-surface displayed glutamate dehydrogenase

    Energy Technology Data Exchange (ETDEWEB)

    Liang, Bo [Laboratory for Biosensing, Key Laboratory of Biofuels, and Shandong Provinicial Key Laboratory of Energy Genetics, Qingdao Institute of Bioenergy & Bioprocess Technology, Chinese Academy of Sciences, 189 Songling Road, Qingdao 266101 (China); University of Chinese Academy of Sciences, 19A Yuquan Road, Beijing 100049 (China); Zhang, Shu [Laboratory for Biosensing, Key Laboratory of Biofuels, and Shandong Provinicial Key Laboratory of Energy Genetics, Qingdao Institute of Bioenergy & Bioprocess Technology, Chinese Academy of Sciences, 189 Songling Road, Qingdao 266101 (China); Key Laboratory of Marine Chemistry Theory and Technology of Ministry of Education, Ocean University of China, 238 Songling Road, Qingdao 266100 (China); Lang, Qiaolin [Laboratory for Biosensing, Key Laboratory of Biofuels, and Shandong Provinicial Key Laboratory of Energy Genetics, Qingdao Institute of Bioenergy & Bioprocess Technology, Chinese Academy of Sciences, 189 Songling Road, Qingdao 266101 (China); Song, Jianxia; Han, Lihui [Key Laboratory of Marine Chemistry Theory and Technology of Ministry of Education, Ocean University of China, 238 Songling Road, Qingdao 266100 (China); Liu, Aihua, E-mail: liuah@qibebt.ac.cn [Laboratory for Biosensing, Key Laboratory of Biofuels, and Shandong Provinicial Key Laboratory of Energy Genetics, Qingdao Institute of Bioenergy & Bioprocess Technology, Chinese Academy of Sciences, 189 Songling Road, Qingdao 266101 (China); University of Chinese Academy of Sciences, 19A Yuquan Road, Beijing 100049 (China)

    2015-07-16

    Highlights: • E. coli surface-dispalyed Gldh exhibiting excellent enzyme activity and stability. • Sensitive amperometric biosensor for glutamate using Gldh-bacteria and MWNTs. • The glutamate biosensor exhibited high specificity and stability. - Abstract: A novel L-glutamate biosensor was fabricated using bacteria surface-displayed glutamate dehydrogenase (Gldh-bacteria). Here the cofactor NADP{sup +}-specific dependent Gldh was expressed on the surface of Escherichia coli using N-terminal region of ice nucleation protein (INP) as the anchoring motif. The cell fractionation assay and SDS-PAGE analysis indicated that the majority of INP-Gldh fusion proteins were located on the surface of cells. The biosensor was fabricated by successively casting polyethyleneimine (PEI)-dispersed multi-walled carbon nanotubes (MWNTs), Gldh-bacteria and Nafion onto the glassy carbon electrode (Nafion/Gldh-bacteria/PEI-MWNTs/GCE). The MWNTs could not only significantly lower the oxidation overpotential towards NAPDH, which was the product of NADP{sup +} involving in the oxidation of glutamate by Gldh, but also enhanced the current response. Under the optimized experimental conditions, the current–time curve of the Nafion/Gldh-bacteria/PEI-MWNTs/GCE was performed at +0.52 V (vs. SCE) by amperometry varying glutamate concentration. The current response was linear with glutamate concentration in two ranges (10 μM–1 mM and 2–10 mM). The low limit of detection was estimated to be 2 μM glutamate (S/N = 3). Moreover, the proposed biosensor is stable, specific, reproducible and simple, which can be applied to real samples detection.

  18. Detection of Waterborne and Airborne Formaldehyde: From Amperometric Chemosensing to a Visual Biosensor Based on Alcohol Oxidase

    Directory of Open Access Journals (Sweden)

    Sasi Sigawi

    2014-02-01

    Full Text Available A laboratory prototype of a microcomputer-based analyzer was developed for quantitative determination of formaldehyde in liquid samples, based on catalytic chemosensing elements. It was shown that selectivity for the target analyte could be increased by modulating the working electrode potential. Analytical parameters of three variants of the amperometric analyzer that differed in the chemical structure/configuration of the working electrode were studied. The constructed analyzer was tested on wastewater solutions that contained formaldehyde. A simple low-cost biosensor was developed for semi-quantitative detection of airborne formaldehyde in concentrations exceeding the threshold level. This biosensor is based on a change in the color of a solution that contains a mixture of alcohol oxidase from the yeast Hansenula polymorpha, horseradish peroxidase and a chromogen, following exposure to airborne formaldehyde. The solution is enclosed within a membrane device, which is permeable to formaldehyde vapors. The most efficient and sensitive biosensor for detecting formaldehyde was the one that contained alcohol oxidase with an activity of 1.2 U·mL−1. The biosensor requires no special instrumentation and enables rapid visual detection of airborne formaldehyde at concentrations, which are hazardous to human health.

  19. Label-free Electrochemiluminescent Immunosensor for Detection of Prostate Specific Antigen based on Aminated Graphene Quantum Dots and Carboxyl Graphene Quantum Dots

    OpenAIRE

    Dan Wu; Yixin Liu; Yaoguang Wang; Lihua Hu; Hongmin Ma; Guoqin Wang; Qin Wei

    2016-01-01

    Prostate-specific antigen (PSA) was used as the model, an ultrasensitive label-free electrochemiluminescent immunosensor was developed based on graphene quantum dots. Au/Ag-rGO was sythsized and used as electrode material to load a great deal of graphene quantum dots due to the large surface area and excellent electron conductivity. After aminated graphene quantum dots and acarboxyl graphene quantum dots were modified onto the electrode, the ECL intensity was much high using K2S2O8 as coreact...

  20. Amperometric inhibition biosensors based on horseradish peroxidase and gold sononanoparticles immobilized onto different electrodes for cyanide measurements.

    Science.gov (United States)

    Attar, Aisha; Cubillana-Aguilera, Laura; Naranjo-Rodríguez, Ignacio; de Cisneros, José Luis Hidalgo-Hidalgo; Palacios-Santander, José María; Amine, Aziz

    2015-02-01

    New biosensors based on inhibition for the detection of cyanide and the comparison of the analytical performances of nine enzyme biosensor designs by using three different electrodes: Sonogel-Carbon, glassy carbon and gold electrodes were discussed. Three different horseradish peroxidase immobilization procedures with and without gold sononanoparticles were studied. The amperometric measurements were performed at an applied potential of -0.15V vs. Ag/AgCl in 50mM sodium acetate buffer solution pH=5.0. The apparent kinetic parameters (Kmapp, Vmaxapp) of immobilized HRP were calculated in the absence of inhibitor (cyanide) by using caffeic acid, hydroquinone, and catechol as substrates. The presence of gold sononanoparticles enhanced the electron transfer reaction and improved the analytical performance of the biosensors. The HRP kinetic interactions reveal non-competitive binding of cyanide with an apparent inhibition constant (Ki) of 2.7μM and I50 of 1.3μM. The determination of cyanide can be achieved in a dynamic range of 0.1-58.6μM with a detection limit of 0.03μM which is lower than those reported by previous studies. Hence this biosensing methodology can be used as a new promising approach for detecting cyanide.

  1. Amperometric catechol biosensor based on laccase immobilized on nitrogen-doped ordered mesoporous carbon (N-OMC)/PVA matrix

    Science.gov (United States)

    Guo, Meiqing; Wang, Hefeng; Huang, Di; Han, Zhijun; Li, Qiang; Wang, Xiaojun; Chen, Jing

    2014-06-01

    A functionalized nitrogen-containing ordered mesoporous carbon (N-OMC), which shows good electrical properties, was synthesized by the carbonization of polyaniline inside a SBA-15 mesoporous silica template. Based on this, through entrapping laccase onto the N-OMC/polyvinyl alcohol (PVA) film a facilely fabricated amperometric biosensor was developed. Laccase from Trametes versicolor was assembled on a composite film of a N-OMC/PVA modified Au electrode and the electrochemical behavior was investigated. The results indicated that the N-OMC modified electrode exhibits electrical properties towards catechol. The optimum experimental conditions of a biosensor for the detection of catechol were studied in detail. Under the optimal conditions, the sensitivity of the biosensor was 0.29 A*M-1 with a detection limit of 0.31 μM and a linear detection range from 0.39 μM to 8.98 μM for catechol. The calibration curve followed the Michaelis-Menten kinetics and the apparent Michaelis-Menten \\left( K_{M}^{app} \\right) was 6.28 μM. This work demonstrated that the N-OMC/PVA composite provides a suitable support for laccase immobilization and the construction of a biosensor.

  2. Self-assembled CNTs/CdS/dehydrogenase hybrid-based amperometric biosensor triggered by photovoltaic effect.

    Science.gov (United States)

    Tang, Longhua; Zhu, Yihua; Yang, Xiaoling; Sun, Jinjie; Li, Chunzhong

    2008-10-15

    A novel multi-components hybrid material, self-assembled quantum dots (CdS) and glutamate dehydrogenase (GDH) onto multiwall carbon nanotubes (CNTs), was designed for amperometric biosensing system. The zeta-potential and transmission electron microscopy (TEM) analyses confirmed the uniform growth of the CdS/GDH onto carboxyl-functionalized CNTs. Compared with the single CdS, the resulting hybrid material showed more efficient generation of photocurrent upon illumination. The incident light excites CdS and generates charge carriers, and then CNTs facilitates the charge transfer. For dehydrogenase-based biosensor, normally, the cofactor of beta-nicotinamide adenine dinucleotide (NAD(+)) or beta-nicotinamide adenine dinucleotide phosphate (NADP(+)) is necessary. Furthermore, we found the photovoltaic effect of CNTs/CdS/GDH can trigger the dehydrogenase enzymatic reaction in the absence of the NAD(+) or NADP(+) cofactors. The electrochemical experiment results also demonstrate that the cofactor-independent dehydrogenase biosensing system had series attractive characteristics, such as a good sensitivity (11.9 nA/microM), lower detection limit (up to 50 nM), an acceptable reproducibility and stability. These studies aid in understanding the combination of the semiconductor nanohybrids (CNTs/QDs, etc.) and biomolecules (enzymes, etc.), which has potential for the applications in biosensor, biofuel cell, biomedical and other bioelectronics field.

  3. Simultaneous Determination of the Main Peanut Allergens in Foods Using Disposable Amperometric Magnetic Beads-Based Immunosensing Platforms

    Directory of Open Access Journals (Sweden)

    Víctor Ruiz-Valdepeñas Montiel

    2016-06-01

    Full Text Available In this work, a novel magnetic beads (MBs-based immunosensing approach for the rapid and simultaneous determination of the main peanut allergenic proteins (Ara h 1 and Ara h 2 is reported. It involves the use of sandwich-type immunoassays using selective capture and detector antibodies and carboxylic acid-modified magnetic beads (HOOC-MBs. Amperometric detection at −0.20 V was performed using dual screen-printed carbon electrodes (SPdCEs and the H2O2/hydroquinone (HQ system. This methodology exhibits high sensitivity and selectivity for the target proteins providing detection limits of 18.0 and 0.07 ng/mL for Ara h 1 and Ara h 2, respectively, with an assay time of only 2 h. The usefulness of the approach was evaluated by detecting the endogenous content of both allergenic proteins in different food extracts as well as trace amounts of peanut allergen (0.0001% or 1.0 mg/kg in wheat flour spiked samples. The developed platform provides better Low detection limits (LODs in shorter assay times than those claimed for the allergen specific commercial ELISA kits using the same immunoreagents and quantitative information on individual food allergen levels. Moreover, the flexibility of the methodology makes it readily translate to the detection of other food-allergens.

  4. Ultrasensitive Label-free Electrochemical Immunosensor based on Multifunctionalized Graphene Nanocomposites for the Detection of Alpha Fetoprotein

    Science.gov (United States)

    Wang, Yaoguang; Zhang, Yong; Wu, Dan; Ma, Hongmin; Pang, Xuehui; Fan, Dawei; Wei, Qin; Du, Bin

    2017-01-01

    In this work, a novel label-free electrochemical immunosensor was developed for the quantitative detection of alpha fetoprotein (AFP). Multifunctionalized graphene nanocomposites (TB-Au-Fe3O4-rGO) were applied to modify the electrode to achieve the amplification of electrochemical signal. TB-Au-Fe3O4-rGO includes the advantages of graphene, ferroferric oxide nanoparticles (Fe3O4 NPs), gold nanoparticles (Au NPs) and toluidine blue (TB). As a kind of redox probe, TB can produce the electrochemical signal. Graphene owns large specific surface area, high electrical conductivity and good adsorption property to load a large number of TB. Fe3O4 NPs have good electrocatalytic performance towards the redox of TB. Au NPs have good biocompatibility to capture the antibodies. Due to the good electrochemical performance of TB-Au-Fe3O4-rGO, the effective and sensitive detection of AFP was achieved by the designed electrochemical immunosensor. Under optimal conditions, the designed immunosensor exhibited a wide linear range from 1.0 × 10−5 ng/mL to 10.0 ng/mL with a low detection limit of 2.7 fg/mL for AFP. It also displayed good electrochemical performance including good reproducibility, selectivity and stability, which would provide potential applications in the clinical diagnosis of other tumor markers. PMID:28186128

  5. An electrochemical immunosensor for ochratoxin A determination in wines based on a monoclonal antibody and paramagnetic microbeads.

    Science.gov (United States)

    Vidal, Juan C; Bonel, Laura; Ezquerra, Alba; Duato, Patricia; Castillo, Juan R

    2012-06-01

    We report a direct competitive immunosensor for the rapid determination of ochratoxin A (OTA) in wine samples. Magnetic beads (1 ± 0.5 μm diameter) covered with streptavidin were functionalized with a monoclonal antibody against OTA, and then left to incubate in a solution of tracer (ochratoxin conjugated to the enzyme peroxidase) and a range of OTA concentrations (10(-4) to 1,000 ng mL(-1)). After washing and separation steps helped with a magnetic field, a volume of the dispersion was put on screen-printed electrodes under a magnet, and after adding the substrate the p-benzoquinone generated enzymatically was detected by differential-pulse voltammetry. Wine samples (2 mL) were easily prepared simply by adjusting to pH = 7.5 with diluted NaOH and by adding polyvinylpyrrolidone for complexing polyphenols, without any other clean-up or preconcentration steps. The limit of detection for detecting OTA in wines was of 0.11 ± 0.01 ng L(-1), well below the permitted content of the mycotoxin by the European Union (wines were subjected to immunosensor calibrations to study the matrix effects. OTA concentrations measured with the immunosensor were compared with those obtained by high-performance liquid chromatography coupled to fluorescence detection (AOAC official method 2001.01). The OTA levels from two red wines of "Campo de Borja", Spain, ranged from about 0.027 to 0.033 ng mL(-1) of OTA.

  6. A novel sandwiched electrochemiluminescence immunosensor for the detection of carcinoembryonic antigen based on carbon quantum dots and signal amplification.

    Science.gov (United States)

    Li, Nian-Lu; Jia, Li-Ping; Ma, Rong-Na; Jia, Wen-Li; Lu, Yi-Yang; Shi, Sha-Shan; Wang, Huai-Sheng

    2017-03-15

    In this study, a novel sandwiched electrochemiluminescence (ECL) immunosensor for the detection of carcinoembryonic antigen (CEA) was developed. The nanocomposite of polydopamine and Ag nanoparticles (PDA-AgNPs) was prepared by the redox reaction between Ag(+) and dopamine. This nanocomposite not only provided an effective matrix for the immobilization of primary antibody (Ab1) but also enhanced the conductivity of the electrode. Carbon quantum dots (CQDs) were immobilized on the poly(ethylenimine) functionalized graphene oxide (PEI-GO) through amido-bond. Then Au nanoparticles were decorated on the CQDs modified PEI-GO matrix, and the resulted complex AuNPs/CQDs-PEI-GO was introduced to link secondary antibody (Ab2). The CQDs can be connected to the electrode surface through the combination of CEA with Ab1 and Ab2, and then the amplified electrochemiluminescence signal of CQDs was obtained with the synergistic effect of AgNPs, polydopamine, AuNPs and PEI-GO. Under the optimal conditions, the ECL intensity was proportional to the logarithm value of CEA concentration in the linear range from 5pgmL(-1) to 500ngmL(-1) with a detection limit of 1.67pgmL(-1) for CEA detection. The immunosensor was applied for the CEA detection in real samples with satisfactory results. The proposed ECL immunosensor showed good performance with high sensitivity, specificity, reproducibility, stability and will be potential in clinical detection.

  7. Label-free electrochemical immunosensor based on gold-silicon carbide nanocomposites for sensitive detection of human chorionic gonadotrophin.

    Science.gov (United States)

    Yang, Long; Zhao, Hui; Fan, Shuangmei; Deng, Shuangsheng; Lv, Qi; Lin, Jie; Li, Can-Peng

    2014-07-15

    Uniform and highly dispersed gold-silicon carbide (Au@SiC) nanocomposites were prepared via simple way and used for fabrication of label-free electrochemical immunosensor for sensitive detection of human chorionic gonadotrophin (hCG). Using Au@SiC as electrode material and using ferricyanide as mediator, the proposed immunosensor provides a simple and economic method with higher sensitivity and a wider concentration range for detection of hCG. Under the optimal condition, the approach provided a good linear response range from 0.1 to 5 IU/L and 5 to 1000 IU/L with a low detection limit of 0.042 IU/L. The immunosensor showed good selectivity, acceptable stability and reproducibility. Satisfactory results were obtained for determination of hCG in human serum samples. The proposed method provides a promising platform of clinical immunoassay for other biomolecules. In addition, the bio-functionalization of SiC combined with other nanomaterials will provide promising approach for electrochemical sensing and biosensing platform.

  8. Ultrasensitive electrochemical immunosensor based on horseradish peroxidase (HRP)-loaded silica-poly(acrylic acid) brushes for protein biomarker detection.

    Science.gov (United States)

    Zhao, Yan; Zheng, Yiqun; Kong, Rongmei; Xia, Lian; Qu, Fengli

    2016-01-15

    We report an ultrasensitive electrochemical immunosensor designed for the detection of protein biomarkers using horseradish peroxidase (HRP)-loaded silica-poly(acrylic acid) brushes (SiO2-SPAABs) as labels. HRP could be efficiently and stably accommodated in the three-dimensional architecture of the SiO2-SPAABs and the SiO2-SPAABs-HRP exhibited high catalytic performance towards o-phenylenediamine (OPD) oxidation in the presence of H2O2, which resulted in significant differential pulse voltammetric (DPV) response change and color change. Using human IgG (HIgG) as a model analyte, a sandwich-type immunosensor was constructed. In particular, graphene oxide (GO) and SiO2-SPAABs-HRP were used to immobilize capture antibody (Ab1) and bind a layer of detection antibody (Ab2), respectively. The current biosensor exhibited a good linear response of HIgG from 100pg/mL to 100μg/mL with a detection limit of 50pg/mL (S/N=5). The sensitivity was 6.70-fold higher than the conventional enzyme-linked immunosorbent assays. The immunosensor results were validated through the detection of HIgG in serum samples.

  9. Nanocomposites of gold nanoparticles and graphene oxide towards an stable label-free electrochemical immunosensor for detection of cardiac marker troponin-I.

    Science.gov (United States)

    Liu, Guozhen; Qi, Meng; Zhang, Yin; Cao, Chaomin; Goldys, Ewa M

    2016-02-25

    A stable label-free amperometric immunosensor is presented based on gold nanoparticles and graphene oxide nanocomposites for detection of cardiac troponin-I in the early diagnosis of myocardial infarction. For designing of the sensing platform, firstly the nanocomposites based on GO and AuNPs were prepared and anchored on electrode surfaces. The formed nanocomposites provided a platform with big surface area for loading anti-cTnI capture antibody, and worked as a bridge for fast electron transfer subsequently increased the sensitivity. Moreover, the linkages between AuNP, GO, and electrodes were based on covalent bonding by aryldiazonium salt coupling chemistry, which favors the stability of the sensing interface. Finally, the anti-cTnI detection antibody was immobilized on GO tailored with ferrocene molecules, functioning as the signal reporter for the detection of cTnI. The modification process was monitored using electrochemistry, SEM, XPS. The herein immunosensor demonstrates a good selectivity and high sensitivity against human-cTnI, and is capable of detecting cTnI at concentrations as low as 0.05 ng mL(-1), which is 100 times lower than that possible by conventional methods. It is potential to design the portable sensing platform based on AuNPs and GO nanocomposites for future point-of-care diagnostics.

  10. Label-free Detection of Influenza Viruses using a Reduced Graphene Oxide-based Electrochemical Immunosensor Integrated with a Microfluidic Platform

    Science.gov (United States)

    Singh, Renu; Hong, Seongkyeol; Jang, Jaesung

    2017-02-01

    Reduced graphene oxide (RGO) has recently gained considerable attention for use in electrochemical biosensing applications due to its outstanding conducting properties and large surface area. This report presents a novel microfluidic chip integrated with an RGO-based electrochemical immunosensor for label-free detection of an influenza virus, H1N1. Three microelectrodes were fabricated on a glass substrate using the photolithographic technique, and the working electrode was functionalized using RGO and monoclonal antibodies specific to the virus. These chips were integrated with polydimethylsiloxane microchannels. Structural and morphological characterizations were performed using X-ray photoelectron spectroscopy and scanning electron microscopy. Electrochemical studies revealed good selectivity and an enhanced detection limit of 0.5 PFU mL‑1, where the chronoamperometric current increased linearly with H1N1 virus concentration within the range of 1 to 104 PFU mL‑1 (R2 = 0.99). This microfluidic immunosensor can provide a promising platform for effective detection of biomolecules using minute samples.

  11. Label-free Detection of Influenza Viruses using a Reduced Graphene Oxide-based Electrochemical Immunosensor Integrated with a Microfluidic Platform

    Science.gov (United States)

    Singh, Renu; Hong, Seongkyeol; Jang, Jaesung

    2017-01-01

    Reduced graphene oxide (RGO) has recently gained considerable attention for use in electrochemical biosensing applications due to its outstanding conducting properties and large surface area. This report presents a novel microfluidic chip integrated with an RGO-based electrochemical immunosensor for label-free detection of an influenza virus, H1N1. Three microelectrodes were fabricated on a glass substrate using the photolithographic technique, and the working electrode was functionalized using RGO and monoclonal antibodies specific to the virus. These chips were integrated with polydimethylsiloxane microchannels. Structural and morphological characterizations were performed using X-ray photoelectron spectroscopy and scanning electron microscopy. Electrochemical studies revealed good selectivity and an enhanced detection limit of 0.5 PFU mL−1, where the chronoamperometric current increased linearly with H1N1 virus concentration within the range of 1 to 104 PFU mL−1 (R2 = 0.99). This microfluidic immunosensor can provide a promising platform for effective detection of biomolecules using minute samples. PMID:28198459

  12. An Amperometric Immunosensor Based on a Polyelectrolyte/ Gold Magnetic Nanoparticle Supramolecular Assembly—Modified Electrode for the Determination of HIV p24 in Serum

    Directory of Open Access Journals (Sweden)

    Minjun Ni

    2010-07-01

    Full Text Available Unmanaged sudden withdrawal from the excessive consumption of alcohol (ethanol adversely alters neuronal integrity in vulnerable brain regions such as the cerebellum, hippocampus, or cortex. In addition to well known hyperexcitatory neurotransmissions, ethanol withdrawal (EW provokes the intense generation of reactive oxygen species (ROS and the activation of stress-responding protein kinases, which are the focus of this review article. EW also inflicts mitochondrial membranes/membrane potential, perturbs redox balance, and suppresses mitochondrial enzymes, all of which impair a fundamental function of mitochondria. Moreover, EW acts as an age-provoking stressor. The vulnerable age to EW stress is not necessarily the oldest age and varies depending upon the target molecule of EW. A major female sex steroid, 17β-estradiol (E2, interferes with the EW-induced alteration of oxidative signaling pathways and thereby protects neurons, mitochondria, and behaviors. The current review attempts to provide integrated information at the levels of oxidative signaling mechanisms by which EW provokes brain injuries and E2 protects against it.

  13. Development of a novel biosensor based on a polypyrrole-dodecylbenzene sulphonate (PPy-DBS) film for the determination of amperometric cholesterol.

    Science.gov (United States)

    Özer, Bayram Oğuz; Çete, Servet

    2017-06-01

    Herein a novel amperometric biosensor based on a conducting polymer with anionic dopant modified electrode was successfully developed for detection of cholesterol. Polypyrrole is deposited on a platinum surface and the sodium dodecylbenzene sulphonate (DBS) ion-doped polypyrrole film was electrochemically prepared by scanning the electrode potential between -0.8 and +0.8 V at a scan rate of 20 mV/s. The present electrochemical biosensor was optimized in terms of working potential, number of cycles, concentrations of monomer, and anionic dopant. Cholesterol oxidase (ChOx) was physically entrapped in PPy-DBS to construct an amperometric cholesterol biosensor. Amperometric determination is based on the electrochemical detection of H2O2 generated in the enzymatic reaction of cholesterol. Kinetic parameters, operational and storage stabilities, pH, and temperature dependencies were determined. Km and Imax were calculated as 0.11 μM and 0.967 nM/min, respectively. The operational stability results showed that 90.0% of the response current was retained after 30 activity assays. Morphology of electrodes was characterized by SEM and AFM. Additionally, contact angle measurements were made with 1 μL water of polymer film and enzyme electrode. As a result, the cholesterol biosensor suggested in this study is easy to prepare and is highly cost-effective. This composite (PPy-DBS) can supply a biocompatible and electrochemical microenvironment for immobilization of the enzyme, making this material a good candidate for the fabrication of highly sensitive and selective cholesterol biosensors.

  14. A Nafion®-based co-planar electrode amperometric sensor for methanol determination in the gas phase

    Indian Academy of Sciences (India)

    K Wallgren; S Sotiropoulos

    2009-09-01

    A co-planar electrode device, fabricated with all electrodes (working, counter and reference) on the same face of a Nafion® polymer electrolyte membrane, is proposed for the amperometric detection of gaseous methanol using Pt as the working electrode. Clear voltammetry is obtained for methanol oxidation from its vapours in equilibrium with methanol aqueous solutions, both in the absence and presence of oxygen in the gas stream. Using an appropriate pulse sequence to keep the indicator electrode active, methanol vapours in the 1-13 Torr partial pressure range (in equilibrium with methanol aqueous solutions in the 1-10% w/w concentration range) could be determined, in a constant potential, amperometric mode. The methanol detector could be operated both in a nitrogen stream and (in what is essential for practical applications) in an air atmosphere too, with estimated detection limits of 1.2 and 1.4 Torr respectively.

  15. Amperometric urea biosensor based on urease and electropolymerized toluidine blue dye as a pH-sensitive redox probe.

    Science.gov (United States)

    Vostiar, Igor; Tkac, Jan; Sturdik, Ernest; Gemeiner, Peter

    2002-05-15

    The electropolymerized toluidine blue film deposited on the glassy carbon electrode show amperometrically detectable pH sensitivity. This feature of polytoluidine blue (PTOB) film was used for a construction of an amperometric urea biosensor. We have observed a linear shift of the formal redox potential with increasing pH value between 4 and 8 giving the slope of 81 mV(Delta) pH(-1). Polytoluidine blue film has had a significantly increased stability and higher electrochemical activity compared to the adsorbed monomeric dye. The polytoluidine blue urea biosensor has been operating at a working potential of -200 mV vs. SCE. The sensitivity of the biosensor was 980 nA mM(-1) cm(-2). The biosensor showed linearity in concentration range up to 0.8 mM with the detection limit of 0.02 mM (S/N=3).

  16. Microfluidic immunosensor with micromagnetic beads coupled to carbon-based screen-printed electrodes (SPCEs) for determination of Botrytis cinerea in tissue of fruits.

    Science.gov (United States)

    Fernández-Baldo, Martín A; Messina, Germán A; Sanz, Maria I; Raba, Julio

    2010-11-10

    A wide range of plant species, including economically important crops such as vegetables, ornamentals, bulbs, and fundamentally fruits, can be affected by gray mold caused by the fungal pathogen Botrytis cinerea . This paper describes the development of a microfluidic immunosensor with micromagnetic beads (MMBs) coupled to carbon-based screen-printed electrodes (SPCEs) for the rapid and sensitive quantification of B. cinerea in apple (Red Delicious), table grape (pink Moscatel), and pear (William's) tissues. The detection of B. cinerea was carried out using a competitive immunoassay method based on the use of purified B. cinerea antigens immobilized on 3-aminopropyl-modified MMBs. The total assay time was 40 min, and the calculated detection limit was 0.008 μg mL(-1). Moreover, the intra- and interassay coefficients of variation were below 7%. The developed method allowed detects B. cinerea even in asymptomatic fruits and promises to be particularly useful for application in the agricultural industry.

  17. Construction of label-free electrochemical immunosensor on mesoporous carbon nanospheres for breast cancer susceptibility gene

    Energy Technology Data Exchange (ETDEWEB)

    Fan, Haixia; Zhang, Yong; Wu, Dan; Ma, Hongmin; Li, Xiaojing; Li, Yan; Wang, Huan; Li, He; Du, Bin [Key Laboratory of Chemical Sensing and Analysis in Universities of Shandong, School of Chemistry and Chemical Engineering, University of Jinan, Jinan 250022 (China); Wei, Qin, E-mail: sdjndxwq@163.com [Key Laboratory of Chemical Sensing and Analysis in Universities of Shandong, School of Chemistry and Chemical Engineering, University of Jinan, Jinan 250022 (China)

    2013-04-03

    Highlights: ► The immunosensor is designed to determine breast cancer susceptibility gene. ► Mesoporous carbon nanospheres (MCN) have great adsorption capacity. ► MCN could enhance the electroactivity of toluidine blue. ► Room temperature ionic liquid should increase the electrochemical signal. -- Abstract: In this contribution, mesoporous carbon nanospheres (MCN) were used to fabricate a label-free electrochemical immunosensor for breast cancer susceptibility gene (BRCAl). The detection platform was constructed by conjugation of anti-BRCA1 on glassy carbon electrodes which were modified by mesoporous carbon nanospheres–toluidine blue nanocomposite (MCN–TB)/room temperature ionic-liquid (RTIL) composited film. TB was adsorbed onto MCN and acted as a redox probe. The electroactivity of TB was greatly enhanced in the presence of MCN. The good conductivity of MCN and BMIM·BF{sub 4} could promote the electron transfer and thus enhance the detection sensitivity. Moreover, the large surface area of MCN and the protein-binding properties of BMIM·BF{sub 4} could greatly increase the antibody loading. The specific antibody–antigen immunoreaction on the electrode surface resulted in a decrease of amperometric signal of the electrode. Under optimized conditions, the amperometric signal decreased linearly with BRCAl concentration in the range of 0.01–15 ng mL{sup −1} with a low detection limit of 3.97 pg mL{sup −1}. The immunosensor exhibits high sensitivity, good selectivity and stability.

  18. Simple and suitable immunosensor for β-lactam antibiotics analysis in real matrixes: milk, serum, urine.

    Science.gov (United States)

    Merola, Giovanni; Martini, Elisabetta; Tomassetti, Mauro; Campanella, Luigi

    2015-03-15

    The anti-penicillin G was conjugated to avidin-peroxidase and biotin to obtain immunogen and competitor which were then used to develop a competitive immunosensor assay for the detection of penicillin G and other β-lactam antibiotics, with Kaff values of the order of 10(8) M(-1). The new immunosensor appears to afford a number of advantages in terms of sensitivity, possibility of "in situ" analysis, but especially of simplicity and lower costs, compared with other existing devices, or different chemical instrumental methods reported in the literature and used for the analysis of β-lactam compounds. Satisfactory results were found in the analysis of real matrixes and good recoveries were obtained by applying the standard addition method to spiked milk, urine, serum and drug samples. The new device uses an amperometric electrode for hydrogen peroxide as transducer, the BSA-penicillin G immobilized on polymeric membrane overlapping the amperometric transducer and the peroxidase enzyme as marker. It proved to be highly sensitive, inexpensive and easily reproducible; LOD was of the order of 10(-11)M. Lastly, the new immunosensor displayed low selectivity versus the entire class of β-lactam antibiotics and higher selectivity toward other classes of non-β-lactam antibiotics.

  19. Sensitive Immunosensor for Cancer Biomarker Based on Dual Signal Amplification Strategy of Graphene Sheets and Multi-Enzyme Functionalized Carbon Nanospheres

    Energy Technology Data Exchange (ETDEWEB)

    Du, Dan; Zou, Zhexiang; Shin, Yongsoon; Wang, Jun; Wu, Hong; Engelhard, Mark H.; Liu, Jun; Aksay, Ilhan A.; Lin, Yuehe

    2010-03-30

    A novel electrochemical immunosensor for sensitive detection of cancer biomarker α fetoprotein (AFP) is described that uses a graphene sheet sensor platform and functionalized carbon nanospheres (CNSs) labeling with horseradish peroxidase-secondary antibodies (HRP-Ab2). Greatly enhanced sensitivity for the cancer biomarker is based on a dual signal amplification strategy: first, the synthesized CNSs yielded a homogeneous and narrow size distribution, which allowed several binding events of HRP-Ab2 on each nanosphere. Enhanced sensitivity was achieved by introducing the multi-bioconjugates of HRP-Ab2-CNSs onto the electrode surface through sandwich immunoreactions. Secondly, functionalized graphene sheets used for the biosensor platform increased the surface area to capture a large amount of primary antibodies (Ab1), thus amplifying the detection response. This amplification strategy is a promising platform for clinical screening of cancer biomarkers and point-of-care diagnostics.

  20. A Surface Plasmon Resonance-Based Immunosensors for Sensitive Detection of Heroin

    Science.gov (United States)

    Wu, Zhong-cheng; Chen, Wen-ge; Wang, Lian-chao; Ge, Yu; Yu, Cheng-duan; Fang, Ting-jian

    2000-12-01

    A simple technique for sensitive detection of heroin based on surface-plasmon-resonance has been theoretically and experimentally investigated. The experiment was realized by using an anti-MO monoclonal antibody and a morphine (MO)-bovine serum albumin (MO-BSA) conjugate (antigen). The reason for using MO-BSA in the detection of heroine was also discussed. MO-BSA was immobilized on a gold thin film of SPR sensor chip by physical adsorption. The configuration of the device is allowed to be further miniaturized, which is required for the construction of a portable SPR device in the application of in-situ analysis.

  1. A Piezoelectric Immunosensor Based on Agglutination Reaction with Amplification of Silica Nanoparticles

    Institute of Scientific and Technical Information of China (English)

    JIN,Xiao-Yong; JIN,Xue-Fang; DING,Yan-Jun; JIANG,Jian-Hui; SHEN,Guo-Li; Ru-Qin

    2008-01-01

    A simple piezoelectric immunoagglutination assay technique with antibody-modified nanoparticles has been developed for direct quantitative detection of protein. The proposed technique is based on the specific agglutination of goat anti-hlgG-coated silica (or gold) nanoparticles in the presence of human immunoglobulin G (hlgG), which causes a frequency change and is monitored by a piezoelectric device. The antibody modified on the probe surface would combine with antibody-coated nanoparticles in the presence of antigen (hlgG) when the surface agglutination reaction took place, which couples both the mass effect and viscoelastic effect acting on the probe. The results indi-cate that the background interference can be substantially minimized and the probe signal can be observably multi-plied. In addition, the surface of the modified probe and that after combining the complex of immunoagglutination were imaged by scanning electronic microscopy (SEM). Moreover, an optimization of assay medium composition with the addition of poly(ethylene glycol) (PEG) serving as immunoagglutination enhancer and sodium chloride to control the ion-strength was investigated. The frequency responses of the immunoagglutination assay were found to correlate well with the hlgG concentration with a detection limit of 84 ng.mL-1.

  2. An amperometric penicillin biosensor with enhanced sensitivity based on co-immobilization of carbon nanotubes, hematein, and {beta}-lactamase on glassy carbon electrode

    Energy Technology Data Exchange (ETDEWEB)

    Chen Bi; Ma Ming [Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research (Ministry of Education of China), College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha 410081 (China); Su Xiaoli, E-mail: xsu@hunnu.edu.cn [Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research (Ministry of Education of China), College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha 410081 (China)

    2010-07-26

    An amperometric penicillin biosensor with enhanced sensitivity was successfully developed by co-immobilization of multi-walled carbon nanotubes (MWCNTs), hematein, and {beta}-lactamase on glassy carbon electrode using a layer-by-layer assembly technique. Under catalysis of the immobilized enzyme, penicillin was hydrolyzed, decreasing the local pH. The pH change was monitored amperometrically with hematein as a pH-sensitive redox probe. MWCNTs were used as an electron transfer enhancer as well as an efficient immobilization matrix for the sensitivity enhancement. The effects of immobilization procedure, working potential, enzyme quantity, buffer concentration, and sample matrix were investigated. The biosensor offered a minimum detection limit of 50 nM (19 {mu}g L{sup -1}) for penicillin V, lower than those of the conventional pH change-based biosensors by more than two orders of magnitude. The electrode-to-electrode variation of the response sensitivity was 7.0% RSD.

  3. Micromachined Amperometric Nitrate Sensor

    OpenAIRE

    Dohyun Kim; Ira Goldberg; Jack Judy

    2003-01-01

    A nitrate-sensing system that consists of a micromachined sensor substrate, nitrate-permeable membrane, integrated microfluidic channels, and standard fluidic connectors has been designed, fabricated, assembled, and tested. Our microsensor was designed for in-situ monitoring of nitrate concentrations in ground water. A silver electrode was patterned for amperometric nitrate detection. An electrochemically oxidized silver electrode was used as a reference electrode. Microfluidic channels were ...

  4. A novel amperometric biosensor based on gold nanoparticles-mesoporous silica composite for biosensing glucose

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    We report a novel bienzyme biosensor based on the assembly of the glucose oxidase (GOD) and horseradish peroxidase (HRP) onto the gold nanoparticles encapsulated mesoporous silica SBA-15 composite (AuNPs-SBA-15). Electrochemical behavior of the bienzyme bioconjugates biosensor is studied by cyclic voltammetry and electrochemical impedance spectroscopy. The results indicate that the presence of mesoporous AuNPs-SBA-15 greatly enhanced the protein loadings, accelerated interfacial electron transfer of HRP and the electroconducting surface, resulting in the realization of direct electrochemistry of HRP. Owing to the electrocatalytic effect of AuNPs-SBA-15 composite, the biosensor exhibits a sensitive response to H2O2 generated from enzymatic reactions. Thus the bienzyme biosensor could be used for the detection of glucose without the addition of any mediator. The detection limit of glucose was 0.5 μM with a linear range from 1 to 48 μM.

  5. Electrophoretically deposited multiwalled carbon nanotube based amperometric genosensor for E.coli detection

    Science.gov (United States)

    Bhardwaj, Hema; Solanki, Shipra; Sumana, Gajjala

    2016-04-01

    This work reports on a sensitive and selective genosensor fabrication method for Escherichia coli (E.coli) detection. The functionalized multiwalled carbon nanotubes (MWCNT) synthesized via chemical vapour deposition have been deposited electrophoretically onto indium tin oxide coated glass surface and have been utilized as matrices for the covalent immobilization of E.coli specific probe oligonucleotide that was identified from the 16s rRNA coding region of the E.coli genome. This fabricated functionalized MWCNT based platform sought to provide improved fundamental characteristics to electrode interface in terms of electro-active surface area and diffusion coefficient. Electrochemical cyclic voltammetry revealed that this genosensor exhibits a linear response to complementary DNA in the concentration range of 10-7 to 10-12 M with a detection limit of 1×10-12 M.

  6. A novel amperometric biosensor based on gold nanoparticles-mesoporous silica composite for biosensing glucose

    Institute of Scientific and Technical Information of China (English)

    ZHANG JingJing; ZHU JunJie

    2009-01-01

    We report a novel bienzyme bioseneor based on the assembly of the glucose oxidase (GOD) and horseradish peroxidase (HRP) onto the gold nanoparticles encapsulated mesoporous silica SBA-15 composite (AuNPs-SBA-15). Electrochemical behavior of the bienzyme bioconjugatse biosensor is studied by cyclic voltammetry and electrochemical impedance spectroscopy. The results indicate that the presence of mesoporous AuNPs-SBA-15 greatly enhanced the protein Ioadings, accelerated inter-facial electron transfer of HRP and the electroconducting surface, resulting in the realization of direct electrochemistry of HRP. Owing to the electrocatalytic effect of AuNPs-SBA-15 composite, the biosen-sor exhibits a sensitive response to H2O2 generated from enzymatic reactions. Thus the bienzyme biosensor could be used for the detection of glucose without the addition of any mediator. The detec-tion limit of glucose was 0.5 μM with a linear range from 1 to 48 μM.

  7. Determination of L- and D-fucose using amperometric electrodes based on diamond paste.

    Science.gov (United States)

    Stefan-van Staden, Raluca-Ioana; Nejem, R'afat Mahmoud; van Staden, Jacobus Frederick; Aboul-Enein, Hassan Y

    2012-02-21

    Monocrystalline diamond (natural diamond, synthetic-1 and synthetic-2) based electrochemical electrodes were designed for the analysis of L- and D-fucose. Response characteristics of the electrochemical electrodes were determined using cyclic voltammetry and differential pulse voltammetry (DPV). L-fucose was determined using DPV with electrodes based on natural diamond, synthetic-1 and synthetic-2, respectively, at 240 mV using NaCl as the electrolyte (pH 3.0); at 160 mV using KNO(3) (pH 10.0) and at 80 mV using KCl as the electrolyte (pH 10.0) while D-fucose was analyzed at 120 mV using KCl as the electrolyte (pH 1.0); at 140 mV using KNO(3) as the electrolyte (pH 1.0) and at 160 mV using NaNO(3) as the electrolyte (pH 3.0). The linear concentration ranges for L-fucose were between 10(-13) and 10(-9) mol L(-1) (natural diamond), 10(-11) and 10(-8) mol L(-1) (synthetic-1) and 10(-6) and 10(-3) mol L(-1) (synthetic-2) with detection limits of 10(-14), 10(-12) and 10(-8) mol L(-1) magnitude order, respectively. For D-fucose, the linear concentration ranges were 10(-6) to 10(-3) mol L(-1) (natural diamond), 10(-5) to 10(-3) mol L(-1) (synthetic-1) and 10(-9) to 10(-3) mol L(-1) (synthetic-2) with detection limits of 10(-7), 10(-7) and 10(-10) mol L(-1) magnitude order, respectively. The sensors were used for the assay of L-fucose in serum and urine samples.

  8. A novel amperometric sensor for peracetic acid based on a polybenzimidazole-modified gold electrode

    Energy Technology Data Exchange (ETDEWEB)

    Hua, Mu-Yi, E-mail: huamy@mail.cgu.edu.t [Green Research Center, Department of Chemical and Materials Engineering, Chang Gung University, 259 Wen-Hwa 1st Rd., Kuei-Shan, Tao-Yuan 33302, Taiwan (China); Biosensor Group, Biomedical Engineering Research Center, Chang Gung University, 259 Wen-Hwa 1st Rd., Kuei-Shan, Tao-Yuan 33302, Taiwan (China); Chen, Hsiao-Chien [Green Research Center, Department of Chemical and Materials Engineering, Chang Gung University, 259 Wen-Hwa 1st Rd., Kuei-Shan, Tao-Yuan 33302, Taiwan (China); Biosensor Group, Biomedical Engineering Research Center, Chang Gung University, 259 Wen-Hwa 1st Rd., Kuei-Shan, Tao-Yuan 33302, Taiwan (China); Tsai, Rung-Ywan [Electronics and Optoelectronics Research Laboratories, Industrial Technology Research Institute, 195, Sec. 4, Chung Hsing Rd., Hsinchu 31040, Taiwan (China); Lin, Yu-Chen [Green Research Center, Department of Chemical and Materials Engineering, Chang Gung University, 259 Wen-Hwa 1st Rd., Kuei-Shan, Tao-Yuan 33302, Taiwan (China); Biosensor Group, Biomedical Engineering Research Center, Chang Gung University, 259 Wen-Hwa 1st Rd., Kuei-Shan, Tao-Yuan 33302, Taiwan (China)

    2011-04-30

    We have developed a peracetic acid (PAA) sensor based on a polybenzimidazole-modified gold (PBI/Au) electrode. Fourier transform infrared and X-ray photoelectron spectroscopy indicated that PAA oxidized 69.4% of the imine in PBI to form PBI N-oxide, increasing the electrochemical reduction current during cyclic voltammetry. The chemical oxidation of the PBI/Au electrode by PAA, followed by its electrochemical reduction, allowed PAA to be detected directly and consecutively by assessing its reduction current. The PAA sensor had a broad linear detection range (3.1 {mu}M-1.5 mM) and a rapid response time (3.9 s) at an applied potential of -0.3 V. Potentially interfering substances, such as hydrogen peroxide, acetic acid, and oxygen, had no effect on the ability of the probe to detect PAA, indicating high selectivity of the probe. Furthermore, the detection range, response time, and sensitivity of the sensor could all be improved by modification of the smooth planar electrode surface to a porous three-dimensional configuration. When compared to the analytical characteristics of other PAA sensors operating under optimal conditions, the three-dimensional PBI/Au electrode offers a rapid detection time, a usable linear range, and a relatively low detection limit.

  9. Love wave immunosensor for the detection of carbaryl pesticide.

    Science.gov (United States)

    Rocha-Gaso, María-Isabel; García, José-Vicente; García, Pablo; March-Iborra, Carmen; Jiménez, Yolanda; Francis, Laurent-Alain; Montoya, Angel; Arnau, Antonio

    2014-09-03

    A Love Wave (LW) immunosensor was developed for the detection of carbaryl pesticide. The experimental setup consisted on: a compact electronic characterization circuit based on phase and amplitude detection at constant frequency; an automated flow injection system; a thermal control unit; a custom-made flow-through cell; and Quartz /SiO2 LW sensors with a 40 μm wavelength and 120 MHz center frequency. The carbaryl detection was based on a competitive immunoassay format using LIB-CNH45 monoclonal antibody (MAb). Bovine Serum Albumin-CNH (BSA-CNH) carbaryl hapten-conjugate was covalently immobilized, via mercaptohexadecanoic acid self-assembled monolayer (SAM), onto the gold sensing area of the LW sensors. This immobilization allowed the reusability of the sensor for at least 70 assays without significant signal losses. The LW immunosensor showed a limit of detection (LOD) of 0.09 μg/L, a sensitivity of 0.31 μg/L and a linear working range of 0.14-1.63 μg/L. In comparison to other carbaryl immunosensors, the LW immunosensor achieved a high sensitivity and a low LOD. These features turn the LW immunosensor into a promising tool for applications that demand a high resolution, such as for the detection of pesticides in drinking water at European regulatory levels.

  10. Love Wave Immunosensor for the Detection of Carbaryl Pesticide

    Directory of Open Access Journals (Sweden)

    María-Isabel Rocha-Gaso

    2014-09-01

    Full Text Available A Love Wave (LW immunosensor was developed for the detection of carbaryl pesticide. The experimental setup consisted on: a compact electronic characterization circuit based on phase and amplitude detection at constant frequency; an automated flow injection system; a thermal control unit; a custom-made flow-through cell; and Quartz /SiO2 LW sensors with a 40 μm wavelength and 120 MHz center frequency. The carbaryl detection was based on a competitive immunoassay format using LIB-CNH45 monoclonal antibody (MAb. Bovine Serum Albumin-CNH (BSA-CNH carbaryl hapten-conjugate was covalently immobilized, via mercaptohexadecanoic acid self-assembled monolayer (SAM, onto the gold sensing area of the LW sensors. This immobilization allowed the reusability of the sensor for at least 70 assays without significant signal losses. The LW immunosensor showed a limit of detection (LOD of 0.09 μg/L, a sensitivity of 0.31 μg/L and a linear working range of 0.14–1.63 μg/L. In comparison to other carbaryl immunosensors, the LW immunosensor achieved a high sensitivity and a low LOD. These features turn the LW immunosensor into a promising tool for applications that demand a high resolution, such as for the detection of pesticides in drinking water at European regulatory levels.

  11. Urea biosensor based on amperometric pH-sensing with hematein as a pH-sensitive redox mediator.

    Science.gov (United States)

    Pizzariello, A; Stredanský, M; Stredanská, S; Miertus, S

    2001-05-30

    The natural dye hematein in water solution was used as a pH-sensitive redox-active mediator for amperometric pH-sensing. The electrochemical characteristics were studied using cyclic voltammetry and chronoamperometry. Several types of urea biosensors were constructed with urease on the surface of platinum and graphite composite electrodes or in the bulk of the graphite composite. They were used for the amperometric urea determination at a working potential of 0 mV (versus SCE) using 0.5 mM hematein. Detection limits and response linearity was in the micromolar range depending on the biosensor type, concentration and pH of buffers used. An interference study of various cations, anions, and substances, which may be present in real samples demonstrated good selectivity for the determination of urea. The biosensors showed good operational (>3 h) and storage (>3 months) stability. The results of urea determination in blood and urine obtained by biosensor correlated well with those obtained by a spectrophotometric reference method.

  12. Amperometric sulfide detection using Coprinus cinereus peroxidase immobilized on screen printed electrode in an enzyme inhibition based biosensor.

    Science.gov (United States)

    Savizi, Iman Shahidi Pour; Kariminia, Hamid-Reza; Ghadiri, Mohammad; Roosta-Azad, Reza

    2012-05-15

    In the present work, an amperometric inhibition biosensor for the determination of sulfide has been fabricated by immobilizing Coprinus cinereus peroxidase (CIP) on the surface of screen printed electrode (SPE). Chitosan/acrylamide was applied for immobilization of peroxidase on the working electrode. The amperometric measurement was performed at an applied potential of -150 mV versus Ag/AgCl with a scan rate of 100 mV in the presence of hydroquinone as electron mediator and 0.1M phosphate buffer solution of pH 6.5. The variables influencing the performance of sensor including the amount of substrate, mediator concentration and electrolyte pH were optimized. The determination of sulfide can be achieved in a linear range of 1.09-16.3 μM with a detection limit of 0.3 μM. Developed sensor showed quicker response to sulfide compared to the previous developed sulfide biosensors. Common anions and cations in environmental water did not interfere with sulfide detection by the developed biosensor. Cyanide interference on the enzyme inhibition caused 43.25% error in the calibration assay which is less than the amounts reported by previous studies. Because of high sensitivity and the low-cost of SPE, this inhibition biosensor can be successfully used for analysis of environmental water samples.

  13. Amperometric bienzyme glucose biosensor based on carbon nanotube modified electrode with electropolymerized poly(toluidine blue O) film

    Energy Technology Data Exchange (ETDEWEB)

    Wang Wenju [Department of Chemistry, Hong Kong Baptist University, Kowloon Tong (Hong Kong); Wang Fang [Department of Chemistry, Hong Kong Baptist University, Kowloon Tong (Hong Kong)] [Department of Chemistry, Wuhan University, Wuhan 430072 (China); Yao Yanli [Department of Chemistry, Hong Kong Baptist University, Kowloon Tong (Hong Kong); Hu Shengshui [Department of Chemistry, Wuhan University, Wuhan 430072 (China); Shiu, Kwok-Keung, E-mail: kkshiu@hkbu.edu.h [Department of Chemistry, Hong Kong Baptist University, Kowloon Tong (Hong Kong)

    2010-09-30

    The amperometric bienzyme glucose biosensor utilizing horseradish peroxidase (HRP) and glucose oxidase (GOx) immobilized in poly(toluidine blue O) (PTBO) film was constructed on multi-walled carbon nanotube (MWNT) modified glassy carbon electrode. The HRP layer could be used to analyze hydrogen peroxide with toluidine blue O (TBO) mediators, while the bienzyme system (HRP + GOx) could be utilized for glucose determination. Glucose underwent biocatalytic oxidation by GOx in the presence of oxygen to yield H{sub 2}O{sub 2} which was further reduced by HRP at the MWNT-modified electrode with TBO mediators. In the absence of oxygen, glucose oxidation proceeded with electron transfer between GOx and the electrode mediated by TBO moieties without H{sub 2}O{sub 2} production. The bienzyme electrode offered high sensitivity for amperometric determination of glucose at low potential, displaying Michaelis-Menten kinetics. The bienzyme glucose biosensor displayed linear response from 0.1 to 1.2 mM with a sensitivity of 113 mA M{sup -1} cm{sup -2} at an applied potential of -0.10 V in air-saturated electrolytes.

  14. A novel antibody–antigen based impedimetric immunosensor for low level detection of HER2 in serum samples of breast cancer patients via modification of a gold nanoparticles decorated multiwall carbon nanotube-ionic liquid electrode

    Energy Technology Data Exchange (ETDEWEB)

    Arkan, Elham [Nano Drug Delivery Research Center, Kermanshah University of Medical Sciences, Kermanshah (Iran, Islamic Republic of); Saber, Reza [Department of Medical Nanotechnology, School of Advanced Medical Technologies, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Research Center for Science and Technology in Medicine, Imam Khomeini Hospital, Tehran (Iran, Islamic Republic of); Karimi, Ziba [Department of Chemistry, Payame Noor University, P.O. Box 19395-3697, Tehran (Iran, Islamic Republic of); Shamsipur, Mojtaba, E-mail: mshamsipur@yahoo.com [Department of Chemistry, Razi University, Kermanshah (Iran, Islamic Republic of)

    2015-05-18

    Highlights: • Design of a novel impedimetric immunosensor for detection of HER2 in serum samples. • Use of a multiwall carbon nanotube-ionic liquid electrode modified with AuNPs as a base. • Immobilization of monoclonal HER2 antibody on AuNPs/MWCILE using 1,6-hexanedithiol as a cross linker. • Achieving linear dynamic range and limit of detection of 10–110 ng mL{sup −1} and 7.4 ng mL{sup −1}, respectively. • Method development and validation and application to assay of HER2 in biological fluids. - Abstract: A highly sensitive impedimetric immunosensor based on a gold nanoparticles/multiwall carbon nanotube-ionic liquid electrode (AuNPs/MW-CILE) was developed for the determination of human epidermal growth factor receptor 2 (HER2). Gold nanoparticles were used to enhance the extent of immobilization and to retain the immunoactivity of the antibody Herceptin on the electrode. Cyclic voltammetry and electrochemical impedance spectroscopy were employed for characterization of various layers coated onto the AuNPs/MW-CILE. The impedance measurements at different steps were based on the charge transfer kinetics of the [Fe(CN){sub 6}]{sup 3−/4−} redox pair. The immobilization of antibody and the corresponding antigen–antibody interaction at the electrode surface altered the interfacial electron transfer. The interactions of antibody with various concentrations of antigen were also monitored via the change of impedance response. The results showed that the charge transfer resistance increases linearly with increasing concentrations of HER2 antigen. The linear range and limit of detection were found as 10–110 ng mL{sup −1} and 7.4 ng mL{sup −1}, respectively. The sensitivity and specificity of the immunosensor were validated. The results showed that the prepared immunosensor is a useful tool for screening of trace amounts of HER2 in serum samples of breast cancer patients.

  15. Flow-Injection Amperometric Determination of Tacrine based on Ion Transfer across a Water–Plasticized Polymeric Membrane Interface

    Directory of Open Access Journals (Sweden)

    C. Rueda

    2007-07-01

    Full Text Available A flow-injection pulse amperometric method for determining tacrine, based onion transfer across a plasticized poly(vinyl chloride (PVC membrane, was developed. Afour-electrode potentiostat with ohmic drop compensation was used, while a flow-throughcell incorporated the four electrodes and the membrane, which containedtetrabutylammonium tetraphenylborate. The influence of the applied potential and of theflow-injection variables on the determination of tacrine was studied. In the selectedconditions, a linear relationship between peak height and tacrine concentration was foundup to 4x10-5M tacrine. The detection limit was 1x10-7M. Good repeatability was obtained.Some common ions and pharmaceutical excipients did not interfere.

  16. Development of an amperometric glucose biosensor based on the immobilization of glucose oxidase in an ormosil-PVA matrix onto a Prussian Blue modified electrode

    Institute of Scientific and Technical Information of China (English)

    CHEN HaiLing; ZHAO Li; CHEN Xi; ZHUANG ZhiXia; WANG XiaoRu

    2009-01-01

    An amperometric glucose biosensor was developed based on the Immobilization of glucose oxidase in the organically modified silicate(ormosil)-polyvinyl acetate(PVA)matrix onto a Prussian Blue(PB)-modified glassy carbon electrode.A higher stability PB-modified electrode was prepared by the electrochemical deposition of FeCl3,K3[Fe(CN)6]and ethylenediamine tetrsacetic acid(EDTA)under cyclic voltammetric(CV)conditions.The effects of the potential range of CV conditions,electrolyte cations,applied potential,pH,temperature and co-existing substances were investigated.The detection limit of the glucose biosensor was 8.1 μmol·L-1(S/N=3)with a linear range from 20 μmol·L-1 to 2 mmol·L-1(R=0.9965).The biosensor presented a fast response and good selectivity.Additionally,excellent reproducibility and stability of the biosensor were observed.

  17. The sandwich-type electrochemiluminescence immunosensor for α-fetoprotein based on enrichment by Fe3O4-Au magnetic nano probes and signal amplification by CdS-Au composite nanoparticles labeled anti-AFP.

    Science.gov (United States)

    Zhou, Hankun; Gan, Ning; Li, Tianhua; Cao, Yuting; Zeng, Saolin; Zheng, Lei; Guo, Zhiyong

    2012-10-09

    A novel and sensitive sandwich-type electrochemiluminescence (ECL) immunosensor was fabricated on a glassy carbon electrode (GCE) for ultra trace levels of α-fetoprotein (AFP) based on sandwich immunoreaction strategy by enrichment using magnetic capture probes and quantum dots coated with Au shell (CdS-Au) as the signal tag. The capture probe was prepared by immobilizing the primary antibody of AFP (Ab1) on the core/shell Fe(3)O(4)-Au nanoparticles, which was first employed to capture AFP antigens to form Fe(3)O(4)-Au/Ab1/AFP complex from the serum after incubation. The product can be separated from the background solution through the magnetic separation. Then the CdS-Au labeled secondary antibody (Ab2) as signal tag (CdS-Au/Ab2) was conjugated successfully with Fe(3)O(4)-Au/Ab1/AFP complex to form a sandwich-type immunocomplex (Fe(3)O(4)-Au/Ab1/AFP/Ab2/CdS-Au), which can be further separated by an external magnetic field and produce ECL signals at a fixed voltage. The signal was proportional to a certain concentration range of AFP for quantification. Thus, an easy-to-use immunosensor with magnetic probes and a quantum dots signal tag was obtained. The immunosensor performed at a level of high sensitivity and a broad concentration range for AFP between 0.0005 and 5.0 ng mL(-1) with a detection limit of 0.2 pg mL(-1). The use of magnetic probes was combined with pre-concentration and separation for trace levels of tumor markers in the serum. Due to the amplification of the signal tag, the immunosensor is highly sensitive, which can offer great promise for rapid, simple, selective and cost-effective detection of effective biomonitoring for clinical application.

  18. A reagentless electrochemiluminescent immunosensor for apurinic/apyrimidinic endonuclease 1 detection based on the new Ru(bpy){sub 3}{sup 2+}/bi-arginine system

    Energy Technology Data Exchange (ETDEWEB)

    Zhao, Min; Chai, Xi Deng Ya-Qin; Han, Jing; Gui, Guo-Feng; Yuan, Ruo, E-mail: yuanruo@swu.edu.cn; Zhuo, Ying, E-mail: yingzhuo@swu.edu.cn

    2014-10-10

    Highlights: • A reagentless ECL biosensor based on the new Ru(bpy){sub 3}{sup 2+}/bi-arginine system. • The successful preparation of bi-Arg/Au@Fe3O4–rGO as enhancer. • Using the APE-1 as target by the sandwich-type immunoassay format. - Abstract: Apurinic/apyrimidinic endonuclease 1 (APE-1), a kind of multifunctional protein widely-distributed in the body, plays an essential role in the DNA base excision repair and serves as multiple possible roles in the response of human cancer to radiotherapy and chemotherapy. In this work, an ultrasensitive solid-state electrochemiluminescence (ECL) immunosensor is designed to determine APE-1 based on the new Ru(bpy){sub 3}{sup 2+}/bi-arginine system. The bi-arginine (bi-Arg) is decorated on the Au nanoparticles functionalized magnetic Fe{sub 3}O{sub 4}/reduced graphene oxide (bi-Arg/Au@Fe{sub 3}O{sub 4}–rGO) according to the self-assembling and covalent cross-linking interaction to obtain the functionalized nanocomposite of bi-Arg/Au@Fe{sub 3}O{sub 4}–rGO. Herein, the bi-Arg/Au@Fe{sub 3}O{sub 4}–rGO plays not only an amplification label to enhance the ECL signal of Ru(bpy){sub 3}{sup 2+} due to the coreactant of bi-Arg but also an ideal nanocarrier to load numerous secondary antibody. Based on sandwich-type immunoassay format, this proposed method offers a linear range of 1.0 fg mL{sup −1}–5.0 pg mL{sup −1} and an estimated detection limit of 0.3 fg mL{sup −1} for the APE-1. Moreover, the reagentless ECL immunosensor also exhibits high sensitivity, excellent selectivity and good stability, which has greatly potential development and application in clinical diagnostics, immunology and biomedical research.

  19. Study on sensitivities of Love-wave immunosensors based on SiO2/36°YX-LiTaO3 and ZnO/36°YX-LiTaO3 structures

    Science.gov (United States)

    Song, Ming-tai; Zhang, Shu-yi; Fan, Li; Zhou, Feng-mei; Wang, Yan; Shui, Xiu-ji

    2013-09-01

    Love-wave immunosensors based on SiO2/36°YX-LiTaO3 or ZnO/36°YX-LiTaO3 structures coated by special sensitive films have been fabricated and studied experimentally. A theoretical model using the partial wave theory of elastic waves propagating in three-layer structures covered by liquid loads is built up to calculate the sensitivities. To determine the elastic constants of the guiding layers, the phase velocities of the corresponding Love-wave devices are measured by a network analyzer, and the theoretical fitting of the experimental data are carried out. Then the relationships between the sensitivities of the Love-wave immunosensors and normalized thicknesses of the guiding layers are obtained, and there are optimized thicknesses of the guiding layers for getting maximum sensitivities. The experimental measurements and numerical calculations of the sensitivities of the Love-wave immunosensors show that both results are essentially consistent with each other.

  20. Introduction to Biosensors From Electric Circuits to Immunosensors

    CERN Document Server

    Yoon, Jeong-Yeol

    2013-01-01

    Introduction to Biosensors: From Electric Circuits to Immunosensors discusses underlying circuitry of sensors for biomedical and biological engineers as well as biomedical sensing modalities for electrical engineers while providing an applications-based approach to the study of biosensors with over 13 extensive, hands-on labs. The material is presented using a building-block approach, beginning with the fundamentals of sensor design and temperature sensors and ending with more complicated biosensors. This book also: Provides electrical engineers with the specific knowledge they need to understand biological sensing modalities Provides biomedical engineers with a solid background in circuits and systems Includes complete coverage of temperature sensors, electrochemical sensors, DNA and immunosensors, piezoelectric sensors and immunosensing in a micofluidic device Introduction to Biosensors: From Electric Circuits to Immunosensors aims to provide an interdisciplinary approach to biosensors that will be apprecia...

  1. Enhanced sensitivity of self-assembled-monolayer-based SPR immunosensor for detection of benzaldehyde using a single-step multi-sandwich immunoassay.

    Science.gov (United States)

    Gobi, K Vengatajalabathy; Matsumoto, Kiyoshi; Toko, Kiyoshi; Ikezaki, Hidekazu; Miura, Norio

    2007-04-01

    This paper describes the fabrication and sensing characteristics of a self-assembled monolayer (SAM)-based surface plasmon resonance (SPR) immunosensor for detection of benzaldehyde (BZ). The functional sensing surface was fabricated by the immobilization of a benzaldehyde-ovalbumin conjugate (BZ-OVA) on Au-thiolate SAMs containing carboxyl end groups. Covalent binding of BZ-OVA on SAM was found to be dependent on the composition of the base SAM, and it is improved very much with the use of a mixed monolayer strategy. Based on SPR angle measurements, the functional sensor surface is established as a compact monolayer of BZ-OVA bound on the mixed SAM. The BZ-OVA-bound sensor surface undergoes immunoaffinity binding with anti-benzaldehyde antibody (BZ-Ab) selectively. An indirect inhibition immunoassay principle has been applied, in which analyte benzaldehyde solution was incubated with an optimal concentration of BZ-Ab for 5 min and injected over the sensor chip. Analyte benzaldehyde undergoes immunoreaction with BZ-Ab and makes it inactive for binding to BZ-OVA on the sensor chip. As a result, the SPR angle response decreases with an increase in the concentration of benzaldehyde. The fabricated immunosensor demonstrates a low detection limit (LDL) of 50 ppt (pg mL(-1)) with a response time of 5 min. Antibodies bound to the sensor chip during an immunoassay could be detached by a brief exposure to acidic pepsin. With this surface regeneration, reusability of the same sensor chip for as many as 30 determination cycles has been established. Sensitivity has been enhanced further with the application of an additional single-step multi-sandwich immunoassay step, in which the BZ-Ab bound to the sensor chip was treated with a mixture of biotin-labeled secondary antibody, streptavidin and biotin-bovine serum albumin (Bio-BSA) conjugate. With this approach, the SPR sensor signal increased by ca. 12 times and the low detection limit improved to 5 ppt with a total response

  2. Porous silicon optical cavity as an immunosensor platform

    Science.gov (United States)

    Lv, Xiao-Yi; Mo, Jia-Qing; Tu, Yi-Xian; Zhong, Fu-Ru; Jiang, Tao; Jia, Zhen-Hong; Li, Jiang-Wei; Zhang, Fu-Chun

    2010-07-01

    A novel porous silicon based optical Fabry-Perot cavity structure is prepared as a label-free immunosensor platform for detecting antigen-antibody. The lagurus zona pelluciad 3 (LZP3) and the specificity of the polyclonal anti-LZP3 antibodies are employed in our laboratory as the target and the probe, respectively. Firstly, the antibodies are immobilized to the porous silicon optical cavity using silanization and glutaraldehyde (GA) chemistry. And then, after the antigen-antibody reaction, it is monitored that the red shift of the reflection spectrum of the immunosensor increases with the antigen concentration. This research also plays a potential role for the extensive applications in immunoassay.

  3. Sensing Escherichia coli O157:H7 via frequency shift through a self-assembled monolayer based QCM immunosensor

    Institute of Scientific and Technical Information of China (English)

    Li-jiang WANG; Chun-sheng WU; Zhao-ying HU; Yuan-fan ZHANG; Rong LI; Ping WANG

    2008-01-01

    By means of the specific immuno-recognition and ultra-sensitive mass detection, a quartz crystal microbalance (QCM) biosensor for Escherichia coli O157:H7 detection was developed in this work. As a suitable surfactant, 16-mercaptohexadecanoic acid (MHDA) was introduced onto the Au surface of QCM, and then self-assembled with N-hydroxysuccinimide (NHS) raster as a reactive intermediate to provide an active interface for the specific antibody immobilization. The binding of target bacteria with the immobilized antibodies decreased the sensor's resonant frequency, and the frequency shift was correlated to the bacterial concentration. The stepwise assembly of the immunosensor was characterized by means of the electrochemical techniques. Using the immersion-dry-immersion procedure, this QCM biosensor could detect 2.0×102 colony forming units (CFU)/ml E. coli O157:H7. In order to reduce the fabrication time, a polyelectrolyte layer-by-layer self-assembly (LBL-SA) method was adopted for fast construction. Finally, the reproducibility of this biosensor was discussed.

  4. Self-enhanced N-(aminobutyl)-N-(ethylisoluminol) derivative-based electrochemiluminescence immunosensor for sensitive laminin detection using PdIr cubes as a mimic peroxidase

    Science.gov (United States)

    Jiang, Xinya; Wang, Huijun; Wang, Haijun; Zhuo, Ying; Yuan, Ruo; Chai, Yaqin

    2016-04-01

    Herein, a self-enhanced N-(aminobutyl)-N-(ethylisoluminol) (ABEI) derivative-based electrochemiluminescence (ECL) immunosensor was constructed for the determination of laminin (LN) using PdIr cubes as a mimic peroxidase for signal amplification. Initially, PdIr cubes with efficient peroxidase mimicking properties, large specific surface areas, and good stability and uniformity were synthesized. Then, l-cysteine (l-Cys) and ABEI were immobilized on the PdIr cubes to form the self-enhanced ECL nanocomplex (PdIr-l-Cys-ABEI). In this nanocomplex, PdIr cubes, whose catalytic constant is higher than that of horseradish peroxidase (HRP), could effectively catalyze H2O2 decomposition and thus enhance the ECL intensity of ABEI. Moreover, PdIr cubes can be easily modified with functional groups, which make them adaptable to desired supported platforms. On the other hand, l-Cys as a coreactant of ABEI could effectively enhance the luminous efficiency due to the intramolecular ECL reaction which could reduce the energy loss between l-Cys and ABEI by giving a shorter electron transfer distance. The developed strategy combined an ABEI derivative as a self-enhanced ECL luminophore and PdIr cubes as a mimic peroxidase, resulting in a significantly enhanced ECL signal output. Also, the strategy showed high sensitivity and selectivity for LN, which suggested that our new approach could be potentially applied in monitoring different proteins.

  5. Carbon Nanotube-Plasma Polymer-Based Amperometric Biosensors: Enzyme-Friendly Platform for Ultrasensitive Glucose Detection

    Science.gov (United States)

    Muguruma, Hitoshi; Matsui, Yasunori; Shibayama, Yu

    2007-09-01

    An amperometric enzyme biosensor fabricated with carbon nanotubes (CNTs) and plasma-polymerized thin films (PPFs) is reported. A mixture of the enzyme glucose oxidase (GOD) and a CNT film is sandwiched with 10-nm-thick acetonitrile PPFs. Under PPF layer was deposited onto a sputtered gold electrode. To facilitate the electrochemical communication between the CNT layer and GOD, CNT was treated with oxygen plasma. The device with single-walled CNTs showed a sensitivity higher than that of multiwalled CNTs. The glucose biosensor showed ultrasensitivity (a sensitivity of 40 μA mM-1 cm-2, a correlation coefficient of 0.992, a linear response range of 0.025-1.9 mM, a detection limit of 6.2 μM at S/N = 3, +0.8 V vs Ag/AgCl), and a rapid response (< 4 s in reaching 95% of maximum response). This high performance is attributed to the fact that CNTs have excellent electrocatalytic activity and enhance electron transfer, and that PPFs and/or the plasma process for CNTs are the enzyme-friendly platform, i.e., a suitable design of the interface between GOD and CNTs.

  6. Microfluidics and nanoparticles based amperometric biosensor for the detection of cyanobacteria (Planktothrix agardhii NIVA-CYA 116) DNA.

    Science.gov (United States)

    Ölcer, Zehra; Esen, Elif; Ersoy, Aylin; Budak, Sinan; Sever Kaya, Dilek; Yağmur Gök, Mehmet; Barut, Serkan; Üstek, Duran; Uludag, Yildiz

    2015-08-15

    Some of the cyanobacteria produce protease inhibitor oligopeptides such as cyanopeptolins and cause drinking water contamination; hence, their detection has great importance to monitor the well-being of water sources that is used for human consumption. In the current study, a fast and sensitive nucleic acid biosensor assay has been described where cyanopeptolin coding region of one of the cyanobacteria (Planktothrix agardhii NIVA-CYA 116) genome has been used as target for monitoring of the fresh water resources. A biochip that has two sets of Au electrode arrays, each consist of shared reference/counter electrodes and 3 working electrodes has been used for the assay. The biochip has been integrated to a microfluidics system and all steps of the assay have been performed during the reagent flow to achieve fast and sensitive DNA detection. On-line hybridization of the target on to the capture probe immobilized surface resulted in a very short assay duration with respect to the conventional static assays. The binding of the avidin and enzyme modified Au nanoparticles to the biotinylated detection probe and the subsequent injection of the substrate enabled a real-time amperometric measurement with a detection limit of 6×10(-12) M target DNA (calibration curve r(2)=0.98). The developed assay enables fast and sensitive detection of cyanopeptolin producing cyanobacteria from freshwater samples and hence shows a promising technology for toxic microorganism detection from environmental samples.

  7. Amperometric detection of carbohydrates based on the glassy carbon electrode modified with gold nano-flake layer

    Directory of Open Access Journals (Sweden)

    Huy Du Nguyen

    2015-09-01

    Full Text Available An electro-deposition approach was established to incorporate the gold nano-flakes onto the glassy carbon electrode in electrochemical cells (nano-Au/GC/ECCs. Using pulsed amperometric detection (PAD without any gold oxidation for cleaning (non-oxidative PAD, the nano-Au/GC/ECCs were able to maintain their activity for oxidizing of carbohydrates in a normal alkaline medium. The reproducibility of peak area was about 2 relative standard deviation (RSD,% for 6 consecutive injections. A dynamic range of carbohydrates was obtained over a concentration range of 5–80 mg L−1 and the limits of detection (LOD were of 2 mg L−1 for fructose and lactose and 1 mg L−1 for glucose and galactose. Moreover, the nano-Au/GC/ECC using the non-oxidative PAD was able to combine with the internal standard method for determination of lactose in fresh cow milk sample.

  8. Fast Determination of Clenbuterol and Salbutamol in Feed and Meat Products Based on Miniaturized Capillary Electrophoresis with Amperometric Detection

    Institute of Scientific and Technical Information of China (English)

    CHU Qing-Cui; GENG Cheng-Huai; ZHOU Hui; YE Jian-Nong

    2007-01-01

    The fast separation capability of a novel miniaturized capillary electrophoresis with an amperometric detection (μCE-AD) system was demonstrated by determining clenbuterol and salbutamol in real samples.The effects of several factors such as the acidity and concentration of the running buffer,the separation voltage,the applied potential and the injection time on CE-AD were examined and optimized.Under the optimum conditions,the two β-agonists could be baseline separated within 60 s at a separation voltage of 2 kV in a 90 mmol/L H3BO3-Na2B4O7 running buffer (pH 7.4),which was not interfered by ascorbic acid and uric acid.Highly linear response was obtained for above compounds over three orders of magnitude with detection limits ranging from 1.20 × 10-7 to 6.50× 10-8 mol/L (S/N=3).This method was successfully used in the analysis of feed and meat products with relatively simple extraction procedures.

  9. Sensitive amperometric determination of methimazole based on the electrocatalytic effect of rutin/multi-walled carbon nanotube film.

    Science.gov (United States)

    Dorraji, Parisa S; Jalali, Fahimeh

    2015-02-01

    Electrochemical deposition was used to prepare a glassy carbon electrode modified with multi-walled carbon nanotubes and the glycosidic compound, rutin (R/MWCNTs/GCE). Cyclic voltammetry of the modified electrode in aqueous solution (pH8) showed a pair of well-defined, stable and reversible redox peaks with surface confined characteristics. The catechol moiety of rutin produced the voltammetric peaks via a 2 electron, 2 proton mechanism in the range of 0.0-0.4V (vs. Ag/AgCl). The transfer coefficient (α), heterogeneous electron transfer rate constant (ks), and surface concentration (Γ) for R/MWCNTs/GCE were calculated by using the cyclic voltammetric data. The modified electrode showed excellent catalytic activity toward oxidation of methimazole. Fixed-potential amperometry was used for sub-micromolar determination of methimazole at pH8. Linear dependence of anodic current to methimazole concentration was obtained in the range of 0.1-26μM of the drug with a limit of detection at 18nM. The modified electrode retained its initial response for at least 2weeks if stored in dry ambient conditions. The electrode was used for the amperometric determination of methimazole in formulations and spiked blood serum samples, successfully.

  10. A new amperometric glucose biosensor based on screen printed carbon electrodes with rhenium(IV - oxide as a mediator

    Directory of Open Access Journals (Sweden)

    ALBANA VESELI

    2012-11-01

    Full Text Available Rhenium(IV-oxide, ReO2, was used as a mediator for carbon paste (CPE and screen printed carbon (SPCE electrodes for the catalytic amperometric determination of hydro-gen peroxide, whose overpotential for the reduction could be lowered to -0.1 V vs. Ag/AgCl in flow injection analysis (FIA using phosphate buffer (0.1 M, pH=7.5 as a carrier. For hydrogen peroxide a detection limit (3σ of 0.8 mg L-1 could be obtained.ReO2-modified SPCEs were used to design biosensors with a template enzyme, i.e. glucose oxidase, entrapped in a Nafion membrane. The resulting glucose sensor showed a linear dynamic range up to 200 mg L-1 glucose with a detection limit (3σ of 0.6 mg L-1. The repeatability was 2.1 % RSD (n = 5 measurements, the reproducibility 5.4 % (n = 5 sensors. The sensor could be applied for the determination of glucose in blood serum in good agreement with a reference method.

  11. Disposable amperometric biosensors based on xanthine oxidase immobilized in the Prussian blue modified screen-printed three-electrode system

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    The screen-printed three-electrode system was applied to fabricate a new type of disposable amperometric xanthine oxidase biosensor.Carbon-working,carbon-counter and Ag/AgCl reference electrodes were all manually printed on the polyethylene terephthalate substrate forming the screen-printed three-electrode system by the conventional screen-printing process.As a mediator,Prussian blue could not only catalyze the electrochemical reduction of hydrogen peroxide produced from the enzyme reaction,but also keep the favorable potential around 0 V.The optimum operational conditions,including pH,potential and temperature,were investigated.The sensitivities of xanthine and hypoxanthine detections were 13.83 mA/M and 25.56 mA/M,respectively.A linear relationship was obtained in the concentration range between 0.10μM and 4.98μM for xanthine and between 0.50μM and 3.98μM for hypoxanthine.The small Michaelis-menten constant value of the xanthine oxidase biosensor was calculated to be 3.90 μM.The results indicate that the fabricated xanthine oxidase biosensor is effective and sensitive for the detection of xanthine and hypoxanthine.

  12. Construction of amperometric uric acid biosensor based on uricase immobilized on PBNPs/cMWCNT/PANI/Au composite.

    Science.gov (United States)

    Rawal, Rachna; Chawla, Sheetal; Chauhan, Nidhi; Dahiya, Tulika; Pundir, C S

    2012-01-01

    A chitosan-glutaraldehyde crosslinked uricase was immobilized onto Prussian blue nanoparticles (PBNPs) absorbed onto carboxylated multiwalled carbon nanotube (c-MWCNT) and polyaniline (PANI) layer, electrochemically deposited on the surface of Au electrode. The nanohybrid-uricase electrode was characterized by scanning electron microscopic (SEM), Fourier transform infrared spectroscopy (FTIR) and cyclic voltammetry. An amperometric uric acid biosensor was fabricated using uricase/c-MWCNT/PBNPs/Au electrode as working electrode, Ag/AgCl as standard and Pt wire as auxiliary electrode connected through a potentiostat. The biosensor showed optimum response within 4s at pH 7.5 and 40°C, when operated at 0.4V vs. Ag/AgCl. The linear working range for uric acid was 0.005-0.8 mM, with a detection limit of 5 μM. The sensor was evaluated with 96% recovery of added uric acid in sera and 4.6 and 5.4% within and between batch of coefficient of variation respectively and a good correlation (r=0.99) with standard enzymic colorimetric method. This sensor measured uric acid in real serum samples. The sensor lost only 37% of its initial activity after its 400 uses over a period of 7 months, when stored at 4°C.

  13. Application on Gold Nanoparticles-Dotted 4-Nitrophenylazo Graphene in a Label-Free Impedimetric Deoxynivalenol Immunosensor

    Directory of Open Access Journals (Sweden)

    Christopher Edozie Sunday

    2015-02-01

    Full Text Available In this paper, we report a new concept to construct a label-free electrochemical inhibition-based immunosensor for the detection of the mycotoxin deoxynivalenol (DON in cereal samples. The electrochemical impedance spectroscopy of tris(bipyridine ruthenium (II chloride was used as a marker enhanced with gold nanoparticles-dotted 4-nitrophenylazo functionalized graphene (AuNp/G/PhNO2 nanocatalyst mediated in Nafion on a glassy carbon electrode. Under the optimized conditions, the formation of immunocomplexes inhibited electron flow and increased the charge transfer resistance of the sensing interface linearly. The change in impedance was proportional to DON concentrations in the range of 6–30 ng/mL with a sensitivity and detection limit of 32.14 ΩL/ng and 0.3 µg/mL, respectively, which compares favorably with the ELISA result. The proposed sensor had a stability of 80.3%, good precision and selectivity in DON standard solution containing different interfering agents, indicating promising application prospect for this strategy in designing impedimetric, electrochemiluminescent, voltammetric or amperometric sensors.

  14. Label-Free Electrochemiluminescent Immunosensor for Detection of Carcinoembryonic Antigen Based on Nanocomposites of GO/MWCNTs-COOH/Au@CeO₂.

    Science.gov (United States)

    Pang, Xuehui; Li, Jianxiu; Zhao, Yongbei; Wu, Dan; Zhang, Yong; Du, Bin; Ma, Hongmin; Wei, Qin

    2015-09-01

    A high-sensitivity electrochemiluminescence (ECL) sensor was conducted to detect carcinoembryonic antigen (CEA). Nanocomposites of graphene oxide/carboxylated multiwall carbon nanotubes/gold/cerium oxide nanoparticles (GO/MWCNTs-COOH/Au@CeO2) were used as antibody carriers and sensing platforms to modify on glassy carbon electrodes (GCE). CeO2 nanoparticles were first exploited as an ECL luminescent material and the possible ECL mechanism was proposed in this work. GO/MWCNTs-COOH was used as a loading matrix for CeO2 nanoparticles because of the superior conductivity and large specific surface area. Au nanoparticles were further deposited on this matrix to attach anti-CEA and enhance the sensitivity of immunosensor. The proposed sensing platform showed excellent cathodic ECL performance and sensitive response to CEA. The effects of experimental conditions on the ECL performance were investigated. The proposed immunosensor showed the broad linear range (0.05-100 ng/mL) and the low detection limit (LOD, 0.02 ng/mL, signal-to-noise ratio = 3) according to the selected experimental conditions. The excellent analysis performance for determination of CEA in the human serum samples simplied this immunosensor displayed high sensitivity and excellent repeatability. More importantly, this conducted immunosensor broadens the use scope of CeO2 nanoparticles.

  15. Electrochemical Detection of Fluoroquinolone Antibiotics in Milk Using a Magneto Immunosensor

    Directory of Open Access Journals (Sweden)

    Daniel G. Pinacho

    2014-08-01

    Full Text Available An amperometric magneto-immunosensor (AMIS for the detection of residues of fluoroquinolone antibiotics in milk samples is described for the first time. The immunosensor presented combines magnetic beads biomodified with an antibody with a broad recognition profile of fluoroquinolones, a haptenized enzyme and a magnetic graphite–epoxy composite (m-GEC electrode. After the immunochemical reaction with specific enzyme tracer, the antibody biomodified magnetic beads are easily captured by an electrode made of graphite-epoxy composite containing a magnet, which also acts as transducer for the electrochemical detection. In spite of the complexity of milk, the use of magnetic beads allows elimination of potential interferences caused by the matrix components; hence the AMIS could perform quantitative measurements, directly in these samples, without any additional sample cleanup or extraction step. The immunosensor is able to detect up to seven different fluoroquinolones far below the MRLs defined by the UE for milk; for example ciprofloxacin is detected directly in milk with an IC50 of 0.74 µg/L and a LOD of 0.009 µg/L. This strategy offers great promise for rapid, simple, cost-effective, and on-site analysis fluoroquinolones in complex samples.

  16. Amperometric Biosensor Based on Diamine Oxidase/Platinum Nanoparticles/Graphene/Chitosan Modified Screen-Printed Carbon Electrode for Histamine Detection.

    Science.gov (United States)

    Apetrei, Irina Mirela; Apetrei, Constantin

    2016-03-24

    This work describes the development and optimization studies of a novel biosensor employed in the detection and quantification of histamine in freshwater fish samples. The proposed biosensor is based on a modified carbon screen-printed electrode with diamineoxidase, graphene and platinum nanoparticles, which detects the hydrogen peroxide formed by the chemical process biocatalysed by the enzyme diamine oxidase and immobilized onto the nanostructurated surface of the receptor element. The amperometric measurements with the biosensor have been implemented in buffer solution of pH 7.4, applying an optimal low potential of +0.4 V. The novel biosensor shows high sensitivity (0.0631 μA·μM), low detection limit (2.54 × 10(-8) M) and a broad linear domain from 0.1 to 300 μM. The applicability in natural complex samples and the analytical parameters of this enzyme sensor have been performed in the quantification of histamine in freshwater fish. An excellent correlation among results achieved with the developed biosensor and results found with the standard method for all freshwater fish samples has been achieved.

  17. Development of an amperometric glucose biosensor based on the immobilization of glucose oxidase in an ormosil-PVA matrix onto a Prussian Blue modified electrode

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    An amperometric glucose biosensor was developed based on the immobilization of glucose oxidase in the organically modified silicate(ormosil)-polyvinyl acetate(PVA) matrix onto a Prussian Blue(PB)-modified glassy carbon electrode.A higher stability PB-modified electrode was prepared by the electrochemical deposition of FeCl3,K3[Fe(CN)6] and ethylenediamine tetraacetic acid(EDTA) under cyclic voltammetric(CV) conditions.The effects of the potential range of CV conditions,electrolyte cations,applied potential,pH,temperature and co-existing substances were investigated.The detection limit of the glucose biosensor was 8.1 μmol·L-1(S/N = 3) with a linear range from 20 μmol·L-1 to 2 mmol·L-1(R = 0.9965).The biosensor presented a fast response and good selectivity.Additionally,excellent reproducibility and stability of the biosensor were observed.

  18. A Biosensor Based on Immobilization of Horseradish Peroxidase in Chitosan Matrix Cross-linked with Glyoxal for Amperometric Determination of Hydrogen Peroxide

    Directory of Open Access Journals (Sweden)

    Gui-Xiang Wang

    2005-05-01

    Full Text Available An amperometric biosensor for hydrogen peroxide (H2O2 was developed viaan easy and effective enzyme immobilization method with the “sandwich” configuration:ferrocene-chitosan: HRP: chitosan-glyoxal using a glassy carbon electrode as the basicelectrode. In order to prevent the loss of immobilized HRP under optimized conditions,the biosensor surface was cross-linked with glyoxal. Ferrocene was selected andimmobilized on the glassy carbon electrode surface as a mediator. The fabricationprocedure was systematically optimized to improve the biosensor performance. Thebiosensor had a fast response of less than 10 s to H2O2, with a linear range of 3.5×10-5 to1.1×10-3 M, and a detection limit of 8.0×10-6 M based on S/N = 3.

  19. A novel electrochemiluminescent immunosensor based on CdS-coated ZnO nanorod arrays for HepG2 cell detection

    Science.gov (United States)

    Liu, Danqing; Wang, Lei; Ma, Shenghua; Jiang, Zhaohua; Yang, Bin; Han, Xiaojun; Liu, Shaoqin

    2015-02-01

    In this work, the highly oriented CdS-coated-ZnO nanorod arrays have been fabricated. The CdS-coated-ZnO nanorod arrays show high electrochemiluminescence intensity, fast response and good stability. All of the desirable properties spur the development of an ECL immunosensor for the detection of the liver cancer cell line (HepG2 cells). Two successive modification steps of 3-aminopropyltriethoxysilane and gold nanoparticles onto the CdS-coated-ZnO nanorod arrays not only offer the substrates for conjugation of antibody, but also effectively enhance the ECL signal, resulting in production of the high performance ECL immunosensor. The ECL immunosensor exhibits a sensitive response to HepG2 cells in a linear range of 300-10 000 cells mL-1 with a detection limit of 256 cells mL-1. The proposed sensor characteristics of high specificity, good reproducibility and remarkable stability will provide a sensitive, selective, and convenient approach for the clinical detection of cancer cells.In this work, the highly oriented CdS-coated-ZnO nanorod arrays have been fabricated. The CdS-coated-ZnO nanorod arrays show high electrochemiluminescence intensity, fast response and good stability. All of the desirable properties spur the development of an ECL immunosensor for the detection of the liver cancer cell line (HepG2 cells). Two successive modification steps of 3-aminopropyltriethoxysilane and gold nanoparticles onto the CdS-coated-ZnO nanorod arrays not only offer the substrates for conjugation of antibody, but also effectively enhance the ECL signal, resulting in production of the high performance ECL immunosensor. The ECL immunosensor exhibits a sensitive response to HepG2 cells in a linear range of 300-10 000 cells mL-1 with a detection limit of 256 cells mL-1. The proposed sensor characteristics of high specificity, good reproducibility and remarkable stability will provide a sensitive, selective, and convenient approach for the clinical detection of cancer cells

  20. A double signal electrochemical human immunoglobulin G immunosensor based on gold nanoparticles-polydopamine functionalized reduced graphene oxide as a sensor platform and AgNPs/carbon nanocomposite as signal probe and catalytic substrate.

    Science.gov (United States)

    Zhang, Si; Huang, Na; Lu, Qiujun; Liu, Meiling; Li, Haitao; Zhang, Youyu; Yao, Shouzhuo

    2016-03-15

    In this paper, a double signal electrochemical Human immunoglobulin G (HIgG) immunosensor based on AgNPs/carbon nanocomposite (Ag/C NC) as the signal probe and catalytic substrate was developed for fast and sensitive detection of HIgG. The as-prepared AuNPs-PDA-rGO nanocomposite and Ag/C NC were confirmed by UV-vis, Fourier transform infrared spectroscopy, scanning electron microscopy and transmission electron microscopy. Electrochemical impedance spectroscopy, cyclic voltammetry and differential pulse voltammetry were used to investigate the electrochemical properties of the proposed immunosensor. The AuNPs-PDA-rGO nanocomposite can improve the electron transfer rate and capture more Ab1. In the sandwich-type immunoassay process, the Ag/C NC functionalized bioconjugates were captured on HIgG/Ab1/AuNPs-PDA-rGO surface and the electrochemical double-signal strategy was employed. These double electrochemical detection signals were directly monitored the oxidation current originated from Ag/C NC and indirectly detected the reduction current of benzoquinone which was produced from the reaction of H2O2 and HQ by catalysis of Ag/C NC in electrochemical detection of HIgG. Under the optimized conditions, the current responses were changed with the concentrations of HIgG for the proposed immunosensor with wide linear ranges of 0.1 to 100 ngmL(-1) and 0.01-100 ngmL(-1) with the lowest detection concentration of 0.001 ng mL(-1) in the absence and presence of H2O2 and HQ. The double-signal strategy is used for detection of HIgG, and the results came from the two signals were well consistent with each other. The proposed immunosensor was successfully applied in analysis of human IgG in real samples and this strategy may provide a relative simple and effective method for construction of other immunsensors in detection of other biomarkers in clinical medicine.

  1. Assembling Amperometric Biosensors for Clinical Diagnostics

    Directory of Open Access Journals (Sweden)

    Claudia Marina Lagier

    2007-02-01

    Full Text Available Clinical diagnosis and disease prevention routinely require the assessment ofspecies determined by chemical analysis. Biosensor technology offers several benefits overconventional diagnostic analysis. They include simplicity of use, specificity for the targetanalyte, speed to arise to a result, capability for continuous monitoring and multiplexing,together with the potentiality of coupling to low-cost, portable instrumentation. This workfocuses on the basic lines of decisions when designing electron-transfer-based biosensorsfor clinical analysis, with emphasis on the strategies currently used to improve the deviceperformance, the present status of amperometric electrodes for biomedicine, and the trendsand challenges envisaged for the near future.

  2. Trends and perspectives in immunosensors

    OpenAIRE

    Viguier, C.; Crean, C; O'Kennedy, R

    2012-01-01

    Immunosensors are devices that comprise both a biomolecular recognition system, such as an antibody and its corresponding antigen, and a transducer to translate the high affinity and specific binding event into a physical signal. Antibodies are produced by an immunological response to the presence of a foreign substance called an antigen. Antibodies may be immobilised onto a variety of platforms including bulk planar surfaces and nanoparticles by either covalent or adsorption strategies. Diff...

  3. Ultrasensitive potentiometric immunosensor based on SA and OCA techniques for immobilization of HBsAb with colloidal Au and polyvinyl butyral as matrixes.

    Science.gov (United States)

    Yuan, Ruo; Tang, Dianping; Chai, Yaqin; Zhong, Xia; Liu, Yan; Dai, Jianyuan

    2004-08-17

    A novel potentiometric immunosensor for detection of hepatitis B surface antigen (HBsAg) has been developed by means of self-assembly (SA) and opposite-charged adsorption (OCA) techniques to immobilize hepatitis B surface antibody (HBsAb) on a platinum electrode. A cleaned platinum electrode was first pretreated in the presence of 10% HNO3 and 2.5% K2CrO4 solution and held at -1.5 V (vs SCE) for 1 min to make it negatively charged and then immersed in a mixing solution containing hepatitis B surface antibody, colloidal gold (Au), and polyvinyl butyral (PVB). Finally, HBsAb was successfully immobilized onto the surface of the negatively charged platinum electrode modified nanosized gold and PVB sol-gel matrixes. The modified procedure was characterized by electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). The immobilized hepatitis B surface antibody exhibited direct electrochemical behavior toward hepatitis B surface antigen (HBsAg). The performance and factors influencing the performance of the resulting immunosensor were studied in detail. More than 95.7% of the results of the human serum samples obtained by this method were in agreement with those obtained by enzyme-linked immunosorbent assays (ELISAs). The resulting immunosensor exhibited fast potentiometric response (6 months).

  4. Highly sensitive electrochemical immunosensor for the detection of alpha fetoprotein based on PdNi nanoparticles and N-doped graphene nanoribbons.

    Science.gov (United States)

    Li, Na; Ma, Hongmin; Cao, Wei; Wu, Dan; Yan, Tao; Du, Bin; Wei, Qin

    2015-12-15

    An ultrasensitive sandwich-type electrochemical immunosensor was designed for the quantitative detection of alpha fetoprotein (AFP). The β-cyclodextrins functionalized graphene sheets (CD-GS) were used as the sensing matrix for immobilizing adamantine-1-carboxylic acid functionalized primary anti-AFP (ADA-Ab1) and enhanced the electron transfer. PdNi alloy nanoparticles decorated N-doped graphene nanoribbons (PdNi/N-GNRs) were used as labels of secondary anti-AFP (Ab2), and PdNi alloy nanoparticles (PdNi NPs) exhibited high catalytic performance towards the reduction of H2O2. Meanwhile, with good dispersion, large specific surface area and good catalytic performance, N-doped graphene nanoribbons (N-GNRs) significantly amplified the electrochemical signal. Under the optimal conditions, the electrochemical immunosensor exhibited a wide linear range of 0.0001-16 ng/mL with a low detection limit of 0.03 pg/mL. Additionally, the proposed immunosensor showed high specificity, good reproducibility and long-term stability, which have promising application in bioassay analysis.

  5. New competitive dendrimer-based and highly selective immunosensor for determination of atrazine in environmental, feed and food samples: the importance of antibody selectivity for discrimination among related triazinic metabolites.

    Science.gov (United States)

    Giannetto, Marco; Umiltà, Eleonora; Careri, Maria

    2014-01-01

    A new voltammetric competitive immunosensor selective for atrazine, based on the immobilization of a conjugate atrazine-bovine serum albumine on a nanostructured gold substrate previously functionalized with poliamidoaminic dendrimers, was realized, characterized, and validated in different real samples of environmental and food concern. Response of the sensor was reliable, highly selective and suitable for the detection and quantification of atrazine at trace levels in complex matrices such as territorial waters, corn-cultivated soils, corn-containing poultry and bovine feeds and corn flakes for human use. Selectivity studies were focused on desethylatrazine, the principal metabolite generated by long-term microbiological degradation of atrazine, terbutylazine-2-hydroxy and simazine as potential interferents. The response of the developed immunosensor for atrazine was explored over the 10(-2)-10(3) ng mL(-1) range. Good sensitivity was proved, as limit of detection and limit of quantitation of 1.2 and 5 ng mL(-1), respectively, were estimated for atrazine. RSD values <5% over the entire explored range attested a good precision of the device.

  6. An amperometric hydrogen peroxide biosensor based on Co{sub 3}O{sub 4} nanoparticles and multiwalled carbon nanotube modified glassy carbon electrode

    Energy Technology Data Exchange (ETDEWEB)

    Kaçar, Ceren; Dalkiran, Berna; Erden, Pınar Esra, E-mail: erdenpe@gmail.com; Kiliç, Esma

    2014-08-30

    Highlights: • Hydrogen peroxide biosensor was constructed by combining the advantageous properties of MWCNTs and Co{sub 3}O{sub 4}. • Incorporating Co{sub 3}O{sub 4} nanoparticles into MWCNTs/gelatin film increased the electron transfer. • Co{sub 3}O{sub 4}/MWCNTs/gelatin/HRP/Nafion/GCE showed strong anti-interference ability. • Hydrogen peroxide was successfully determined in disinfector with an average recovery of 100.78 ± 0.89. - Abstract: In this work a new type of hydrogen peroxide biosensor was fabricated based on the immobilization of horseradish peroxidase (HRP) by cross-linking on a glassy carbon electrode (GCE) modified with Co{sub 3}O{sub 4} nanoparticles, multiwall carbon nanotubes (MWCNTs) and gelatin. The introduction of MWCNTs and Co{sub 3}O{sub 4} nanoparticles not only enhanced the surface area of the modified electrode for enzyme immobilization but also facilitated the electron transfer rate, resulting in a high sensitivity of the biosensor. The fabrication process of the sensing surface was characterized by scanning electron microscopy (SEM), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Amperometric detection of hydrogen peroxide was investigated by holding the modified electrode at −0.30 V (vs. Ag/AgCl). The biosensor showed optimum response within 5 s at pH 7.0. The optimized biosensor showed linear response range of 7.4 × 10{sup −7}–1.9 × 10{sup −5} M with a detection limit of 7.4 × 10{sup −7}. The applicability of the purposed biosensor was tested by detecting hydrogen peroxide in disinfector samples. The average recovery was calculated as 100.78 ± 0.89.

  7. Towards the conception of an amperometric sensor of L-tyrosine based on Hemin/PAMAM/MWCNT modified glassy carbon electrode

    Energy Technology Data Exchange (ETDEWEB)

    Ma Qiang [College of Chemistry and Material Science, Shandong Agricultural University, Taian 271018, Shandong (China); Ai Shiyun, E-mail: ashy@sdau.edu.c [College of Chemistry and Material Science, Shandong Agricultural University, Taian 271018, Shandong (China); Yin Huanshun [College of Chemistry and Material Science, Shandong Agricultural University, Taian 271018, Shandong (China); Chen Quanpeng; Tang Tiantian [College of Resources and Environment, Shandong Agricultural University, Taian 271018, Shandong (China)

    2010-09-01

    A novel amperometric sensor was fabricated based on the immobilization of hemin onto the poly (amidoamine)/multi-walled carbon nanotube (PAMAM/MWCNT) nanocomposite film modified glassy carbon electrode (GCE). Electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV) and ultraviolet visible (UV-vis) adsorption spectroscopy were used to investigate the possible state and electrochemical activity of the immobilized hemin. In the Hemin/PAMAM/MWCNT nanocomposite film, MWCNT layer possessed excellent inherent conductivity to enhance the electron transfer rate, while the layer of PAMAM greatly enlarged the surface average concentration of hemin ({Gamma}) on the modified electrode. Therefore, the nanocomposite film showed enhanced electrocatalytical activity towards the oxidation of L-tyrosine. The kinetic parameters of the modified electrode were investigated. In pH 7.0 phosphate buffer solution (PBS), the sensor exhibits a wide linear range from 0.1 {mu}M to 28.8 {mu}M L-tyrosine with a detection limit of 0.01 {mu}M and a high sensitivity of 0.31 {mu}A {mu}M{sup -1} cm{sup -2}. In addition, the response time of the L-tyrosine sensor is less than 5 s. The excellent performance of the sensor is largely attributed to the electro-generated high reactive oxoiron (IV) porphyrin (O = Fe{sup IV}-P) which effectively catalyzed the oxidation of L-tyrosine. A mechanism was herein proposed for the catalytic oxidation of L-tyrosine by oxoiron (IV) porphyrin complexes.

  8. 基于碳纳米管修饰玻碳电极的唾液sIgA可再生免疫传感器%Renewable Salivary Secretory Immunoglobulin A(sIgA) Immunosensor Based on Carbon Nanotube Modified Glassy Carbon Electrode

    Institute of Scientific and Technical Information of China (English)

    王艳; 甄生航; 朱洋; 谢国明

    2011-01-01

    A novel amperometric immunosensor based on multi-wall carbon nanotubes (MWNTs)/layer-by-layer(LBL) assembly of gold nanoparticles and thionine( GNPs- THI)/secretory immunoglobulin A( sIgA)/horseradish peroxidase(HRP) was developed for the detection of salivary secretory immunoglobulin A (sIgA). The layer-by-layer(LBL) assembly of gold nanoparticles and thionine (GNPs-THI) was assembled through electrostatic adsorption, Au- S and Au- N covalent bonds.Then the salivary anti - sIgA antibody was adsorbed by the formative monolayer of GNPs. Eventually,the horseradish peroxidase (HRP) was used to block the nonspecific adsorption sites on the modified glassy carbon electrode (GCE) and amplify the current signal at the same time. The immunosensor showed a high sensitivity to salivary sIgA with a detection limit (S/N = 3 ) of 1.5 μg/L. The linear range was 3.0 -350. 0 μg/L with correlation coefficient(r) of 0. 989 7. The response time achieved 95% of the steady state was less than 20 s. Also, the immunosensor had an excellent interference immunity to the interferents of immunoglobulin G ( IgG), α-amylase and albumin, and the interference rates were - 2. 13% , - 2. 85% and - 1.96% , respectively. The antigen - antibody complexes in the measurement were decollemented by 0. 1 moL/L of aminoacetic acid. The bioactivities of antigen and antibody were remained well after decollement and the biosensor could be recycled by aminoacetic acid for eight times. Some influence factors on the biosensor were investigated, such as the concentration of substrate and antibody, incubation time, interferents, temperature and pH. It can be used for accurate and fast detection of salivary slgA to judge the impairment of the mucosal inmune system of human owing to its high sensitivity, excellent specificity and convenient detection, and it can be easily regenerated.%将壳聚糖(CHI)分散的羧基化多壁碳纳米管(MWCNT)吸附到玻碳电极(GCE)表面,形成负电荷界面,利用

  9. A novel amperometric biosensor for superoxide anion based on superoxide dismutase immobilized on gold nanoparticle-chitosan-ionic liquid biocomposite film

    Energy Technology Data Exchange (ETDEWEB)

    Wang Lu; Wen Wei; Xiong Huayu; Zhang Xiuhua; Gu Haoshuang [Ministry of Education Key Laboratory for the Synthesis and Application of Organic Functional Molecules and College of Chemistry and Chemical Engineering, Hubei University, Wuhan 430062 (China); Wang Shengfu, E-mail: wangsf@hubu.edu.cn [Ministry of Education Key Laboratory for the Synthesis and Application of Organic Functional Molecules and College of Chemistry and Chemical Engineering, Hubei University, Wuhan 430062 (China)

    2013-01-03

    Graphical abstract: Schematic representation of the assembly process of SOD/GNPs-CS-IL/GCE. Highlights: Black-Right-Pointing-Pointer SOD was immobilized in gold nanoparticles-chitosan-ionic liquid (GNPs-CS-IL) film. Black-Right-Pointing-Pointer The biosensor was constructed by one-step ultrasonic electrodeposition of GNPs-CS-IL onto GCE. Black-Right-Pointing-Pointer The biosensor showed excellent analytical performance for O{sub 2}{center_dot}{sup -} real-time analysis. - Abstract: A novel superoxide anion (O{sub 2}{center_dot}{sup -}) biosensor is proposed based on the immobilization of copper-zinc superoxide dismutase (SOD) in a gold nanoparticle-chitosan-ionic liquid (GNPs-CS-IL) biocomposite film. The SOD-based biosensor was constructed by one-step ultrasonic electrodeposition of GNP-CS-IL composite onto glassy carbon electrode (GCE), followed by immobilization of SOD on the modified electrode. Surface morphologies of a set of representative films were characterized by scanning electron microscopy. The electrochemical performance of the biosensor was evaluated by cyclic voltammetry and chronoamperometry. A pair of quasi-reversible redox peaks of SOD with a formal potential of 0.257 V was observed at SOD/GNPs-CS-IL/GCE in phosphate buffer solution (PBS, 0.1 M, pH 7.0). The effects of varying test conditions on the electrochemical behavior of the biosensor were investigated. Furthermore, several electrochemical parameters were calculated in detail. Based on the biomolecule recognition of the specific reactivity of SOD toward O{sub 2}{center_dot}{sup -}, the developed biosensor exhibited a fast amperometric response (<5 s), wide linear range (5.6-2.7 Multiplication-Sign 10{sup 3} nM), low detection limit (1.7 nM), and excellent selectivity for the real-time measurement of O{sub 2}{center_dot}{sup -}. The proposed method is promising for estimating quantitatively the dynamic changes of O{sub 2}{center_dot}{sup -} in biological systems.

  10. Label-Free and Multiplex Detection of Antibiotic Residues in Milk Using Imaging Surface Plasmon Resonance-Based Immunosensor

    NARCIS (Netherlands)

    Raz, Sabina Rebe; Bremer, Maria G. E. G.; Haasnoot, Willem; Norde, Willem

    2009-01-01

    Monitoring of antimicrobial drug residues in foods relies greatly on the availability of adequate analytical techniques. Currently, there is a need for a high-throughput screening method with a broad-spectrum detection range. This paper describes the development of a microarray biosensor, based on a

  11. Label-Free and Multiplex Detection of Antibiotic Residues in Milk Using Imaging surface Plasmon Resonance-Based immunosensor

    NARCIS (Netherlands)

    Rebe, S.; Bremer, M.G.E.G.; Haasnoot, W.; Norde, W.

    2009-01-01

    Monitoring of antimicrobial drug residues in foods relies greatly on the availability of adequate analytical techniques. Currently, there is a need for a high-throughput screening method with a broad-spectrum detection range. This paper describes the development of a microarray biosensor, based on a

  12. An Immunosensor Based on Antibody Binding Fragments Attached to Gold Nanoparticles for the Detection of Peptides Derived from Avian Influenza Hemagglutinin H5

    Directory of Open Access Journals (Sweden)

    Urszula Jarocka

    2014-08-01

    Full Text Available This paper concerns the development of an immunosensor for detection of peptides derived from avian influenza hemagglutinin H5. Its preparation consists of successive gold electrode modification steps: (i modification with 1,6-hexanedithiol and gold colloidal nanoparticles; (ii immobilization of antibody-binding fragments (Fab’ of anti-hemagglutinin H5 monoclonal antibodies Mab 6-9-1 via S-Au covalent bonds; and (iii covering the remaining free space on the electrode surfaces with bovine serum albumin. The interactions between Fab’ fragments and hemagglutinin (HA variants have been explored with electrochemical impedance spectroscopy (EIS in the presence of [Fe(CN6]3−/4− as an electroactive marker. The immunosensor was able to recognize three different His-tagged variants of recombinant hemagglutinin from H5N1 viruses: H1 subunit (17–340 residues of A/swan/Poland/305-135V08/2006, the long HA (17–530 residues A/Bar-headed Goose/Qinghai/12/2005 and H1 subunit (1–345 residues of A/Vietnam/1194/2004. The strongest response has been observed for the long variant with detection limit of 2.2 pg/mL and dynamic range from 4.0 to 20.0 pg/mL.

  13. An amperometric biosensor based on acetylcholinesterase immobilized onto iron oxide nanoparticles/multi-walled carbon nanotubes modified gold electrode for measurement of organophosphorus insecticides

    Energy Technology Data Exchange (ETDEWEB)

    Chauhan, Nidhi [Department of Biochemistry, M.D. University, Rohtak 124001, Haryana (India); Pundir, Chandra Shekhar, E-mail: pundircs@rediffmail.com [Department of Biochemistry, M.D. University, Rohtak 124001, Haryana (India)

    2011-09-02

    Graphical abstract: The stepwise amperometric biosensor fabrication process and immobilized acetylcholinesterase inhibition in pesticide solution. Highlights: {center_dot} Constructed a novel composite material using Fe{sub 3}O{sub 4}NP and c-MWCNT at Au electrode for electrocatalysis. {center_dot} The properties of nanoparticles modified electrodes were studied by SEM, FTIR, CVs and EIS. {center_dot} The biosensor exhibited good sensitivity (0.475 mA {mu}M{sup -1}) {center_dot} The half life of electrode was 2 months. {center_dot} The sensor was suitable for trace detection of OP pesticide residues in milk and water. - Abstract: An acetylcholinesterase (AChE) purified from maize seedlings was immobilized covalently onto iron oxide nanoparticles (Fe{sub 3}O{sub 4}NP) and carboxylated multi walled carbon nanotubes (c-MWCNT) modified Au electrode. An organophosphorus (OP) biosensor was fabricated using this AChE/Fe{sub 3}O{sub 4}/c-MWCNT/Au electrode as a working electrode, Ag/AgCl as standard and Pt wire as an auxiliary electrode connected through a potentiostat. The biosensor was based on inhibition of AChE by OP compounds/insecticides. The properties of nanoparticles modified electrodes were studied by scanning electron microscopy (SEM), Fourier transform infrared (FTIR), cyclic voltammograms (CVs) and electrochemical impedance spectroscopy (EIS). The synergistic action of Fe{sub 3}O{sub 4}NP and c-MWCNT showed excellent electrocatalytic activity at low potential (+0.4 V). The optimum working conditions for the sensor were pH 7.5, 35 deg. C, 600 {mu}M substrate concentration and 10 min for inhibition by pesticide. Under optimum conditions, the inhibition rates of OP pesticides were proportional to their concentrations in the range of 0.1-40 nM, 0.1-50 nM, 1-50 nM and 10-100 nM for malathion, chlorpyrifos, monocrotophos and endosulfan respectively. The detection limits were 0.1 nM for malathion and chlorpyrifos, 1 nM for monocrotophos and 10 nM for endosulfan. The

  14. Label-free and multiplex detection of antibiotic residues in milk using imaging surface plasmon resonance-based immunosensor.

    Science.gov (United States)

    Rebe Raz, Sabina; Bremer, Maria G E G; Haasnoot, Willem; Norde, Willem

    2009-09-15

    Monitoring of antimicrobial drug residues in foods relies greatly on the availability of adequate analytical techniques. Currently, there is a need for a high-throughput screening method with a broad-spectrum detection range. This paper describes the development of a microarray biosensor, based on an imaging surface plasmon resonance (iSPR) platform, for quantitative and simultaneous immunodetection of different antibiotic residues in milk. Model compounds from four major antibiotic families: aminoglycosides (Neomycin, Gentamicin, Kanamycin, and Streptomycin), sulfonamides (Sulfamethazine), fenicols (Chloramphenicol), and fluoroquinolones (Enrofloxacin) were detected using a single sensor chip. By multiplexing seven immunoassays in a competitive format, we were able to measure all the target compounds at parts per billion (ppb) levels in buffer and in 10x-diluted milk. The assays for Neomycin, Kanamycin, Streptomycin, Enrofloxacin, and Sulfamethazine were sensitive enough for milk control at maximum residue levels as established in the European Union. The overall performance of the biosensor was determined to be comparable to that of conventional four-channel surface plasmon resonance (SPR)-based biosensors, in terms of assay sensitivity and robustness. Combining the advantages of a SPR sensor and a microarray, utilization of the biosensor described here offers a promising alternative to the existing methods and is highly relevant for multianalyte food profiling.

  15. An enzymatic microreactor based on chaotic micromixing for enhanced amperometric detection in a continuous glucose monitoring application

    NARCIS (Netherlands)

    Moon, B.-U.; Koster, S.; Wientjes, K.J.C.; Kwapiszewski, R.M.; Schoonen, A.J.M.; Westerink, B.H.C.; Verpoorte, E.

    2010-01-01

    The development of continuous glucose monitoring systems is a major trend in diabetes-related research. Small, easy-to-wear systems which are robust enough to function over many days without maintenance are the goal. We present a new sensing system for continuous glucose monitoring based on a micror

  16. A Label-Free Immunosensor for IgG Based on an Extended-Gate Type Organic Field Effect Transistor

    Directory of Open Access Journals (Sweden)

    Tsukuru Minamiki

    2014-09-01

    Full Text Available A novel biosensor for immunoglobulin G (IgG detection based on an extended-gate type organic field effect transistor (OFET has been developed that possesses an anti-IgG antibody on its extended-gate electrode and can be operated below 3 V. The titration results from the target IgG in the presence of a bovine serum albumin interferent, clearly exhibiting a negative shift in the OFET transfer curve with increasing IgG concentration. This is presumed to be due an interaction between target IgG and the immobilized anti-IgG antibody on the extended-gate electrode. As a result, a linear range from 0 to 10 µg/mL was achieved with a relatively low detection limit of 0.62 µg/mL (=4 nM. We believe that these results open up opportunities for applying extended-gate-type OFETs to immunosensing.

  17. Conductometric immunosensors for the detection of staphylococcal enterotoxin B based bio-electrocalytic reaction on micro-comb electrodes.

    Science.gov (United States)

    Chen, Zai-Gang

    2008-06-01

    Staphylococcal enterotoxin B (SEB) is one of many toxins produced by the Gram-positive bacterium Staphylococcal aureus. While SEB is known as the causative agent of certain food poisonings it is also considered abiological select agent. Thus, rapid and accurate identification of SEB during either surveillance or in response to a biothreat is critical to the mitigation of the suspect agent. This report presents a new conductometric immune-biosensor for the detection of SEB based on immobilization of horseradish peroxidase (HRP)-labeled SEB antibody (HRP-anti-SEB) onto nanogold/chitosan-multiwalled carbon nanotube (Au/CTS-MWNT)-functionalized biorecognition interface. The formation of the antibody-antigen complex by a simple one-step immunoreaction between the immobilized HRP-anti-SEB and SEB in sample solution introduced a barrier of electrical communication between the immobilized HRP and the base surface, thus local conductivity variations could be evaluated by the bio-electrocatalytic reaction of HRP in 0.02 M PBS (pH 6.8) containing 0.15 mM H(2)O(2), 0.06 M KI and 0.1 M NaCl. Under optimal conditions, the proposed immune-biosensor exhibited a good conductometric response relative to SEB concentration in a linear range from 0.5 to 83.5 ng/ml with a correlation coefficient of 0.998. The developed immune-biosensor showed an acceptable accuracy, reproducibility and stability. Milk samples spiked with various concentrations of SEB gave an average of 116% recovery of the toxin.

  18. A streptavidin functionalized graphene oxide/Au nanoparticles composite for the construction of sensitive chemiluminescent immunosensor

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Zhanjun, E-mail: zjyang@yzu.edu.cn [Key Laboratory of Environmental Material and Environmental Engineering of Jiangsu Province, College of Chemistry and Chemical Engineering, Yangzhou University, Yangzhou 225002 (China); Luo, Shufen; Li, Juan; Shen, Juan; Yu, Suhua; Hu, Xiaoya [Key Laboratory of Environmental Material and Environmental Engineering of Jiangsu Province, College of Chemistry and Chemical Engineering, Yangzhou University, Yangzhou 225002 (China); Dionysiou, Dionysios D. [Environmental Engineering and Science Program, School of Energy Environmental, Biological and Medical Engineering, University of Cincinnati, Cincinnati, OH 45221-0012 (United States)

    2014-08-11

    Highlights: • A novel streptavidin/GO/AuNPs composite is prepared for immobilizing antibody. • A highly sensitive chemiluminescent immunosensor is constructed for tumor marker. • The immunoassay system shows extremely low detection limit down to picogram level. • This work provides a promising approach for ultrasensitive biosensing applications. - Abstract: In this work, a novel streptavidin functionalized graphene oxide/Au nanoparticles (streptavidin/GO/AuNPs) composite is prepared and for the first time used to construct sensitive chemiluminescent immunosensor for the detection of tumor marker. The streptavidin/GO/AuNPs composite and the immunosensor are characterized using scanning electron microscopy, static water contact angle measurement and electrochemical impedance spectroscopy. The biofunctionalized composite has large reactive surface area and excellent biocompatibility, thus the capture antibody can be efficiently immobilized on its surface based on the highly selective recognition of streptavidin to biotinylated antibody. Using α-fetoprotein (AFP) as a model, the proposed chemiluminescent immunosensor shows a wide linear range from 0.001 to 0.1 ng mL{sup −1} with an extremely low detection limit down to 0.61 pg mL{sup −1}. The resulting AFP immunosensor shows high detection sensitivity, fast assay speed, acceptable detection and fabrication reproducibility, good specificity and stability. The assay results of serum samples with the proposed method are in an acceptable agreement with the reference values. This work provides a promising biofunctionalized nanostructure for sensitive biosensing applications.

  19. Electrochemical impedance immunosensor for the detection of cardiac biomarker Myogobin (Mb) in aqueous solution

    Energy Technology Data Exchange (ETDEWEB)

    Rajesh, E-mail: rajesh_csir@yahoo.com; Sharma, V.; Tanwar, V.K.; Mishra, S.K.; Biradar, A.M.

    2010-11-30

    A label-free, electrochemical impedance immunosensor based on surface modified thin flat gold wire electrode is reported for the quantitative detection of cardiac biomarker Myoglobin in aqueous solution. The protein antibody, ab-Mb, was covalently immobilized through a self assembled monolayer of 11-mercaptoundecanoic acid (MUA) and 3-mercapto propionic acid (MPA) via carbodiimide coupling reaction using N-(3-dimethylaminopropyl)-N'-ethyl carbodiimide hydrochloride (EDC) and N-Hydroxy Succinamide (NHS). The immunosensor (ab-Mb/MUA-MPA/Au) was characterized by electrochemical techniques. The electrochemical performance of the immunosensor was studied by electrochemical impedance spectroscopy. The immunosensor showed an increased electrontransfer resistance on coupling with biomarker protein antigen, ag-Mb, in the presence of a redox probe [Fe (CN){sub 6}]{sup 3-/4-}. The modified Au electrode immunosensor exhibits an electrochemical impedance response to antigen, ag-Mb concentrations in a linear range from 10 ng to 650 ng mL{sup -1} with a lowest detection limit of 5.2 ng mL{sup -1}.

  20. A novel urea amperometric biosensor based on secretion of carnation petal cells modified on a graphite-epoxy composite electrode.

    Science.gov (United States)

    Pang, Chunyan; Zhu, Yongchun; Gao, Hongyan; Dong, Yue; Lu, Jie

    2011-02-21

    A new kind of biosensor for the detection of urea with a high selectivity, sensitivity and wide detection range was designed based on the secretion of carnation petals cells paste covered over a graphite-epoxy composite basic electrode surface. The carnation petal paste from mashed fresh carnation petals was tightly fixed on the basic electrode surface with Teflon thin film to keep it in contact with the electrode surface. Urea in aqueous solution was detected by differential pulse voltammetry based on the oxidation peak current at 0.316 V (vs. SCE) of the secreted species of carnation petal cells during the mashing process, which interacts with urea molecules and results in the decrease of the oxidation peak current. The oxidation peak current decreases linearly with the logarithm of urea concentration in the range of 1.3 × 10(-16)-4.57 × 10(-8) M and 3.4 × 10(-7)-1.3 × 10(-1) M with a detection limit of 7.5 × 10(-16) M. The biosensor was characterized by electrochemistry and fluorescent spectrometry, and applied to the determination of urea in waste water from a river around Shenyang Normal University campus with a recovery of 104.5% (RSD is 5.00%). The presence of larger amounts of ammonium ion and nitrate ion up to the molar ratio of 10(4) do not interfere with the urea detection.

  1. An electrochemical magneto immunosensor (EMIS) for the determination of paraquat residues in potato samples.

    Science.gov (United States)

    Garcia-Febrero, Raul; Valera, Enrique; Muriano, Alejandro; Pividori, M-Isabel; Sanchez-Baeza, Francisco; Marco, M-Pilar

    2013-09-01

    An electrochemical magneto immunosensor for the detection of low concentrations of paraquat (PQ) in food samples has been developed and its performance evaluated in a complex sample such as potato extracts. The immunosensor presented uses immunoreagents specifically developed for the recognition of paraquat, a magnetic graphite-epoxy composite (m-GEC) electrode and biofunctionalized magnetic micro-particles (PQ1-BSAMP) that allow reduction of the potential interferences caused by the matrix components. The amperometric signal is provided by an enzymatic probe prepared by covalently linking an enzyme to the specific antibodies (Ab198-cc-HRP). The use of hydroquinone, as mediator, allows recording of the signal at a low potential, which also contributes to reducing the background noise potentially caused by the sample matrix. The immunocomplexes formed on top of the modified MP are easily captured by the m-GEC, which acts simultaneously as transducer. PQ can be detected at concentrations as low as 0.18 ± 0.09 μg L(-1). Combined with an efficient extraction procedure, PQ residues can be directly detected and accurately quantified in potato extracts without additional clean-up or purification steps, with a limit of detection (90% of the maximum signal) of 2.18 ± 2.08 μg kg(-1), far below the maximum residue level (20 μg kg(-1)) established by the EC. The immunosensor presented here is suitable for on-site analysis. Combined with the use of magnetic racks, multiple samples can be run simultaneously in a reasonable time.

  2. Amperometric uric acid biosensor based on poly(vinylferrocene)-gelatin-carboxylated multiwalled carbon nanotube modified glassy carbon electrode.

    Science.gov (United States)

    Erden, Pınar Esra; Kaçar, Ceren; Öztürk, Funda; Kılıç, Esma

    2015-03-01

    In this study, a new uric acid biosensor was constructed based on ferrocene containing polymer poly(vinylferrocene) (PVF), carboxylated multiwalled carbon nanotubes (c-MWCNT) and gelatin (GEL) modified glassy carbon electrode (GCE). Uricase enzyme (UOx) was immobilized covalently through N-ethyl-N'-(3-dimethyaminopropyl) carbodiimide (EDC) and N-hydroxyl succinimide (NHS) chemistry onto c-MWCNT/GEL/PVF/GCE. The c-MWCNT/GEL/PVF composite was characterized by scanning electron microscopy, cyclic voltammetry and electrochemical impedance spectroscopy. Various experimental parameters such as pH, applied potential, enzyme loading, PVF and c-MWCNT concentration were investigated in detail. Under the optimal conditions the dynamic linear range of uric acid was 2.0×10(-7) M-7.1×10(-4) M (R=0.9993) with the detection limit low to 2.3×10(-8) M. With good selectivity and sensitivity, the biosensor was successfully applied to determine the uric acid in human serum. The results of the biosensor were in good agreement with those obtained from standard method. Therefore, the presented biosensor could be a good promise for practical applications in real samples.

  3. Amperometric biosensor based on Laccase immobilized onto a screen-printed electrode by Matrix Assisted Pulsed Laser Evaporation.

    Science.gov (United States)

    Verrastro, Maria; Cicco, Nunzia; Crispo, Fabiana; Morone, Antonio; Dinescu, Maria; Dumitru, Marius; Favati, Fabio; Centonze, Diego

    2016-07-01

    A Laccase-based biosensor for the determination of phenolic compounds was developed by using Matrix Assisted Pulsed Laser Evaporation as an innovative enzyme immobilization technique. and the deriving biosensor was characterized and applied for the first time. Laccase was immobilized onto different substrates including screen printed carbon electrodes and spectroscopic, morphologic and electrochemical characterizations were carried out. A linear range from 1 to 60μM was achieved working at 5.5pH and -0.2V detection potential vs Ag pseudoreference. The limits of detection and quantification were found to be 1 and 5μM, respectively. A good fabrication reproducibility, stability of response and selectivity toward interferents were also found The potential of the developed biosensor was tested in the determination of total polyphenol content in real matrices (tea infusion, ethanolic extract from Muscari comosum bulbs and aqueous solution of a food supplement from black radish root and artichoke leaves) and the results were compared with those obtained by using the Folin-Ciocalteu method.

  4. Amperometric Biosensor Based on Zirconium Oxide/Polyethylene Glycol/Tyrosinase Composite Film for the Detection of Phenolic Compounds.

    Science.gov (United States)

    Ahmad, Nor Monica; Abdullah, Jaafar; Yusof, Nor Azah; Ab Rashid, Ahmad Hazri; Abd Rahman, Samsulida; Hasan, Md Rakibul

    2016-06-29

    A phenolic biosensor based on a zirconium oxide/polyethylene glycol/tyrosinase composite film for the detection of phenolic compounds has been explored. The formation of the composite film was expected via electrostatic interaction between hexacetyltrimethylammonium bromide (CTAB), polyethylene glycol (PEG), and zirconium oxide nanoparticles casted on screen printed carbon electrode (SPCE). Herein, the electrode was treated by casting hexacetyltrimethylammonium bromide on SPCE to promote a positively charged surface. Later, zirconium oxide was mixed with polyethylene glycol and the mixture was dropped cast onto the positively charged SPCE/CTAB. Tyrosinase was further immobilized onto the modified SPCE. Characterization of the prepared nanocomposite film and the modified SPCE surface was investigated by scanning electron microscopy (SEM), Electrochemical Impedance Spectroscopy (EIS), and Cyclic voltamogram (CV). The developed biosensor exhibits rapid response for less than 10 s. Two linear calibration curves towards phenol in the concentrations ranges of 0.075-10 µM and 10-55 µM with the detection limit of 0.034 µM were obtained. The biosensor shows high sensitivity and good storage stability for at least 30 days.

  5. Amperometric Biosensor Based on Zirconium Oxide/Polyethylene Glycol/Tyrosinase Composite Film for the Detection of Phenolic Compounds

    Science.gov (United States)

    Ahmad, Nor Monica; Abdullah, Jaafar; Yusof, Nor Azah; Ab Rashid, Ahmad Hazri; Abd Rahman, Samsulida; Hasan, Md. Rakibul

    2016-01-01

    A phenolic biosensor based on a zirconium oxide/polyethylene glycol/tyrosinase composite film for the detection of phenolic compounds has been explored. The formation of the composite film was expected via electrostatic interaction between hexacetyltrimethylammonium bromide (CTAB), polyethylene glycol (PEG), and zirconium oxide nanoparticles casted on screen printed carbon electrode (SPCE). Herein, the electrode was treated by casting hexacetyltrimethylammonium bromide on SPCE to promote a positively charged surface. Later, zirconium oxide was mixed with polyethylene glycol and the mixture was dropped cast onto the positively charged SPCE/CTAB. Tyrosinase was further immobilized onto the modified SPCE. Characterization of the prepared nanocomposite film and the modified SPCE surface was investigated by scanning electron microscopy (SEM), Electrochemical Impedance Spectroscopy (EIS), and Cyclic voltamogram (CV). The developed biosensor exhibits rapid response for less than 10 s. Two linear calibration curves towards phenol in the concentrations ranges of 0.075–10 µM and 10–55 µM with the detection limit of 0.034 µM were obtained. The biosensor shows high sensitivity and good storage stability for at least 30 days. PMID:27367738

  6. Photoelectrochemical Immunosensor for Detection of Carcinoembryonic Antigen Based on 2D TiO2 Nanosheets and Carboxylated Graphitic Carbon Nitride.

    Science.gov (United States)

    Wang, Huan; Wang, Yaoguang; Zhang, Yong; Wang, Qi; Ren, Xiang; Wu, Dan; Wei, Qin

    2016-06-06

    Carcinoembryonic antigen (CEA) was used as the model, an ultrasensitive label-free photoelectrochemical immunosensor was developed using 2D TiO2 nanosheets and carboxylated graphitic carbon nitride (g-C3N4) as photoactive materials and ascorbic acid as an efficient electron donor. 2D TiO2 nanosheets was sythsized by surfactant self-assembly method and proved to have higher photoelectrochemical signals than TiO2 nanoparticles. Firstly, carboxylated g-C3N4 could be attached to 2D TiO2 nanosheets through the bond formed between carboxyl group of carboxylated g-C3N4 and TiO2. And the photocurrent of g-C3N4/TiO2 drastically enhances compared to carboxylated g-C3N4 and TiO2. Then, antibody of CEA was bonded to TiO2 through the dentate bond formed between carboxyl group of anti-CEA and TiO2, leading to the decrease of the photocurrents. As proven by PEC experiments and electrochemical impedance spectroscopy (EIS) analysis, the fabrication process of the immunosensor is successful. Under the optimal conditions, the intensity decreased linearly with CEA concentration in the range of 0.01~10 ng/mL. The detection limit is 2.1 pg/mL. The work provides an effective method for the detection of tumor markers and can be extended for the application in food safety and environmental monitoring analysis.

  7. Nanocomposites of gold nanoparticles and graphene oxide towards an stable label-free electrochemical immunosensor for detection of cardiac marker troponin-I

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Guozhen, E-mail: gzliu@mail.ccnu.edu.cn [Key Laboratory of Pesticide and Chemical Biology of Ministry of Education, College of Chemistry, Central China Normal University, Wuhan 430079 (China); ARC Centre of Excellence in Nanoscale Biophotonics (CNBP), Department of Physics and Astronomy, Macquarie University, North Ryde 2109 (Australia); Qi, Meng; Zhang, Yin; Cao, Chaomin [Key Laboratory of Pesticide and Chemical Biology of Ministry of Education, College of Chemistry, Central China Normal University, Wuhan 430079 (China); Goldys, Ewa M. [ARC Centre of Excellence in Nanoscale Biophotonics (CNBP), Department of Physics and Astronomy, Macquarie University, North Ryde 2109 (Australia)

    2016-02-25

    A stable label-free amperometric immunosensor is presented based on gold nanoparticles and graphene oxide nanocomposites for detection of cardiac troponin-I in the early diagnosis of myocardial infarction. For designing of the sensing platform, firstly the nanocomposites based on GO and AuNPs were prepared and anchored on electrode surfaces. The formed nanocomposites provided a platform with big surface area for loading anti-cTnI capture antibody, and worked as a bridge for fast electron transfer subsequently increased the sensitivity. Moreover, the linkages between AuNP, GO, and electrodes were based on covalent bonding by aryldiazonium salt coupling chemistry, which favors the stability of the sensing interface. Finally, the anti-cTnI detection antibody was immobilized on GO tailored with ferrocene molecules, functioning as the signal reporter for the detection of cTnI. The modification process was monitored using electrochemistry, SEM, XPS. The herein immunosensor demonstrates a good selectivity and high sensitivity against human-cTnI, and is capable of detecting cTnI at concentrations as low as 0.05 ng mL{sup −1}, which is 100 times lower than that possible by conventional methods. It is potential to design the portable sensing platform based on AuNPs and GO nanocomposites for future point-of-care diagnostics. - Highlights: • Nanocomposites based on GO and AuNPs were prepared and anchored on the electrode surfaces covalently to form a stable sensing interface. • The anti-cTnI detection antibody was immobilized on GO tailored with ferrocene molecules, functioning as the signal reporter for the detection of cTnI. • The detectable concentration of cTnI is 0.05 ng mL{sup -1} in buffer with the assay time of less than 5 min. • The herein simple and novel approach for fabrication of AuNP and graphene based platform is promising for future fabrication of point-of-care devices.

  8. Simultaneous detection of two breast cancer-related miRNAs in tumor tissues using p19-based disposable amperometric magnetobiosensing platforms.

    Science.gov (United States)

    Torrente-Rodríguez, R M; Campuzano, S; López-Hernández, E; Montiel, V Ruiz-Valdepeñas; Barderas, R; Granados, R; Sánchez-Puelles, J M; Pingarrón, J M

    2015-04-15

    A novel magnetobiosensing approach for the rapid and simultaneous detection of two breast cancer-related miRs (miR-21 and miR-205) is reported. It involves the use of antimiR-21 and antimiR-205 specific probes, chitin-modified magnetic beads (Chitin-MBs), the p19 viral protein as capture bioreceptor and amperometric detection with the H2O2/hydroquinone (HQ) system at dual screen-printed carbon electrodes (SPdCEs). The use of SPdCEs allows the simultaneous independent amperometric readout for each target miR to be measured. The magnetosensor exhibited sensitive and selective detection with dynamic ranges from 2.0 to 10.0nM and detection limits of 0.6nM (6fmol) for both target miRs without any amplification step in less than 2h. The usefulness of the approach was evaluated by detecting the endogenous levels of both target miRs in total RNA (RNAt) extracted from metastatic breast cancer cell lines and human tissues.

  9. Amperometric biosensor based on prussian blue and nafion modified screen-printed electrode for screening of potential xanthine oxidase inhibitors from medicinal plants.

    Science.gov (United States)

    El Harrad, Loubna; Amine, Aziz

    2016-04-01

    A simple and sensitive amperometric biosensor was developed for the screening of potential xanthine oxidase inhibitors from medicinal plants. This biosensor was prepared by immobilization of xanthine oxidase on the surface of prussian blue modified screen-printed electrodes using nafion and glutaraldehyde. The developed biosensor showed a linear amperometric response at an applied potential of +0.05 V toward the detection of hypoxanthine from 5 μM to 45 μM with a detection limit of 0.4 μM (S/N=3) and its sensitivity was found to be 600 mA M(-1) cm(-2). In addition, the biosensor exhibited a good storage stability. The inhibition of xanthine oxidase by allopurinol was studied under the optimized conditions. The linear range of allopurinol concentration is obtained up to 2.5 μM with an estimated 50% of inhibitionI50=1.8 μM. The developed biosensor was successfully applied to the screening of xanthine oxidase inhibitors from 13 medicinal plants belonging to different families. Indeed, Moroccan people traditionally use these plants as infusion for the treatment of gout and its related symptoms. For this purpose, water extracts obtained from the infusion of these plants were used for the experiments. In this work, 13 extracts were assayed and several of them demonstrated xanthine oxidase inhibitory effect, with an inhibition greater than 50% compared to spectrophotometry measurements that only few extracts showed an inhibition greater than 50%.

  10. Amperometric cholesterol biosensor based on the direct electrochemistry of cholesterol oxidase and catalase on a graphene/ionic liquid-modified glassy carbon electrode.

    Science.gov (United States)

    Gholivand, Mohammad Bagher; Khodadadian, Mehdi

    2014-03-15

    Cholesterol oxidase (ChOx) and catalase (CAT) were co-immobilized on a graphene/ionic liquid-modified glassy carbon electrode (GR-IL/GCE) to develop a highly sensitive amperometric cholesterol biosensor. The H2O2 generated during the enzymatic reaction of ChOx with cholesterol could be reduced electrocatalytically by immobilized CAT to obtain a sensitive amperometric response to cholesterol. The direct electron transfer between enzymes and electrode surface was investigated by cyclic voltammetry. Both enzymes showed well-defined redox peaks with quasi-reversible behaviors. An excellent sensitivity of 4.163 mA mM(-1)cm(-2), a response time less than 6s, and a linear range of 0.25-215 μM (R(2)>0.99) have been observed for cholesterol determination using the proposed biosensor. The apparent Michaelis-Menten constant (KM(app)) was calculated to be 2.32 mM. The bienzymatic cholesterol biosensor showed good reproducibility (RSDsanalytical performance for the determination of free cholesterol in human serum samples.

  11. Band-type microelectrodes for amperometric immunoassays

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Ga-Yeon; Chang, Young Wook; Ko, Hyuk [Department of Materials Science and Engineering, Yonsei University, 134 Shinchon-dong, Seodaemun-gu, Seoul, 120-749 (Korea, Republic of); Kang, Min-Jung [Korea Institute of Science and Technology (KIST), Seoul (Korea, Republic of); Pyun, Jae-Chul, E-mail: jcpyun@yonsei.ac.kr [Department of Materials Science and Engineering, Yonsei University, 134 Shinchon-dong, Seodaemun-gu, Seoul, 120-749 (Korea, Republic of)

    2016-07-20

    A band-type microelectrode was made using a parylene-N film as a passivation layer. A circular-type, mm-scale electrode with the same diameter as the band-type microelectrode was also made with an electrode area that was 5000 times larger than the band-type microelectrode. By comparing the amperometric signals of 3,5,3′,5′-tetramethylbenzidine (TMB) samples at different optical density (OD) values, the band-type microelectrode was determined to be 9 times more sensitive than the circular-type electrode. The properties of the circular-type and the band-type electrodes (e.g., the shape of their cyclic voltammograms, the type of diffusion layer used, and the diffusion layer thickness per unit electrode area) were characterized according to their electrode area using the COMSOL Multiphysics software. From these simulations, the band-type electrode was estimated to have the conventional microelectrode properties, even when the electrode area was 100 times larger than a conventional circular-type electrode. These results show that both the geometry and the area of an electrode can influence the properties of the electrode. Finally, amperometric analysis based on a band-type electrode was applied to commercial ELISA kits to analyze human hepatitis B surface antigen (hHBsAg) and human immunodeficiency virus (HIV) antibodies. - Highlights: • A band-type microelectrode was made using a parylene-N film as a passivation layer. • The band-type microelectrode was 14-times more sensitive than circular-type electrode. • The influence of geometry on microelectrode properties was simulated using COMSOL. • The band-type electrode was applied to ELISA kits for hHBsAg and hHIV-antibodies.

  12. Amperometric IFN-γ immunosensors with commercially fabricated PCB sensing electrodes.

    Science.gov (United States)

    Moschou, Despina; Greathead, Louise; Pantelidis, Panagiotis; Kelleher, Peter; Morgan, Hywel; Prodromakis, Themistoklis

    2016-12-15

    Lab-on-a-Chip (LoC) technology has the potential to revolutionize medical Point-of-Care diagnostics. Currently, considerable research efforts are focused on innovative production technologies that will make commercial upscaling of lab-on-chip products financially viable. Printed circuit board (PCB) manufacturing techniques have several advantages in this field. In this paper we focus on transferring a complete IFN-γ enzyme-linked immune-sorbent assay (ELISA) onto a commercial PCB electrochemical biosensing platform, We adapted a commercially available ELISA to detect the enzyme product TMB/H2O2 using amperometry, successfully reproducing the colorimetry-obtained ELISA standard curve. The results demonstrate the potential for the integration of these components into an automated, disposable, electronic ELISA Lab-on-PCB diagnostic platform.

  13. Electrochemical Impedance Immunosensor Based on Self-Assembled Monolayers for Rapid Detection of Escherichia coli O157:H7 with Signal Amplification Using Lectin

    Directory of Open Access Journals (Sweden)

    Zhanming Li

    2015-08-01

    Full Text Available Escherichia coli O157:H7 is a predominant foodborne pathogen with severe pathogenicity, leading to increasing attention given to rapid and sensitive detection. Herein, we propose an impedance biosensor using new kinds of screen-printed interdigitated microelectrodes (SPIMs and wheat germ agglutinin (WGA for signal amplification to detect E. coli O157:H7 with high sensitivity and time-efficiency. The SPIMs integrate the high sensitivity and short response time of the interdigitated electrodes and the low cost of the screen-printed electrodes. Self-assembling of bi-functional 3-dithiobis-(sulfosuccinimidyl-propionate (DTSP on the SPIMs was investigated and was proved to be able to improve adsorption quantity and stability of biomaterials. WGA was further adopted to enhance the signal taking advantage of the abundant lectin-binding sites on the bacteria surface. The immunosensor exhibited a detection limit of 102 cfu·mL−1, with a linear detection range from 102 to 107 cfu·mL−1 (r2 = 0.98. The total detection time was less than 1 h, showing its comparable sensitivity and rapid response. Furthermore, the low cost of one SPIM significantly reduced the detection cost of the biosensor. The biosensor may have great promise in food safety analysis and lead to a portable biosensing system for routine monitoring of foodborne pathogens.

  14. Label-Free 3D Ag Nanoflower-Based Electrochemical Immunosensor for the Detection of Escherichia coli O157:H7 Pathogens

    Science.gov (United States)

    Huang, He; Liu, Minghuan; Wang, Xiangsheng; Zhang, Wenjie; Yang, Da-Peng; Cui, Lianhua; Wang, Xiansong

    2016-11-01

    It is highly desirable to develop a rapid and simple method to detect pathogens. Combining nanomaterials with electrochemical techniques is an efficient way for pathogen detection. Herein, a novel 3D Ag nanoflower was prepared via a biomineralization method by using bovine serum albumin (BSA) as a template. It was adopted as a sensing interface to construct an electrochemical bacteria immunosensor for the rapid detection of foodborne pathogens Escherichia coli ( E. coli) O157:H7. Bacterial antibody was immobilized onto the surface of Ag nanoflowers through covalent conjugation. Electrochemical impedance spectroscopy (EIS) was used to detect and validate the resistance changes, where [Fe(CN)6]3-/4- acted as the redox probe. A linear relation between R et and E. coli concentration was obtained in the E. coli concentration range of 3.0 × 102-3.0 × 108 cfu mL-1. The as-prepared biosensor gave rise to an obvious response to E. coli but had no distinct response to Cronobacter sakazakii, methicillin-resistant Staphylococcus aureus (MRSA), Staphylococcus albus, Lactobacillus easei, and Shigella flexneri, revealing a high selectivity for the detection of the pathogens down to 100 cfu mL-1 in a short time. We believe that this BSA-conjugated 3D Ag nanoflowers could be used as a powerful interface material with good conductivity and biocompatibility for improving pathogen detection and treatment in the field of medicine, environment, and food safety.

  15. A novel immunosensor based on immobilization of hepatitis B surface antibody on platinum electrode modified colloidal gold and polyvinyl butyral as matrices via electrochemical impedance spectroscopy.

    Science.gov (United States)

    Tang, Dianping; Yuan, Ruo; Chai, Yaqin; Dai, Jianyuan; Zhong, Xia; Liu, Yan

    2004-12-01

    Hepatitis B surface antibody (HBsAb) was immobilized to the surface of platinum electrode modified with colloidal gold and polyvinyl butyral (PVB) as matrices to detect hepatitis B surface antigen (HBsAg) via electrochemical impedance spectroscopy (EIS). The electrochemical measurements of cyclic voltammetry and impedance spectroscopy showed that K(4)[Fe(CN)(6)]/K(3)[Fe(CN)(6)] reactions on the platinum electrode surface were blocked due to the procedures of self-assembly of HBsAb-Au-PVB. The binding of a specific HBsAb to HBsAg recognition layer could be detected by measurements of the impedance change. A new strategy was introduced for improving the sensitivity of impedance measurements via the large specific surface area and high surface free energy of Au nanoparticles and the encapsulated effect of polyvinyl butyral. The results showed that this strategy caused dramatic improvement of the detection sensitivity of HBsAg and had good linear response to detect HBsAg in the range of 20-160 ng.ml(-1) with a detection limit of 7.8 ng.ml(-1). Moreover, the studied immunosensor exhibited high sensitivity and long-term stability.

  16. Electrochemical Impedance Immunosensor Based on Self-Assembled Monolayers for Rapid Detection of Escherichia coli O157:H7 with Signal Amplification Using Lectin

    Science.gov (United States)

    Li, Zhanming; Fu, Yingchun; Fang, Weihuan; Li, Yanbin

    2015-01-01

    Escherichia coli O157:H7 is a predominant foodborne pathogen with severe pathogenicity, leading to increasing attention given to rapid and sensitive detection. Herein, we propose an impedance biosensor using new kinds of screen-printed interdigitated microelectrodes (SPIMs) and wheat germ agglutinin (WGA) for signal amplification to detect E. coli O157:H7 with high sensitivity and time-efficiency. The SPIMs integrate the high sensitivity and short response time of the interdigitated electrodes and the low cost of the screen-printed electrodes. Self-assembling of bi-functional 3-dithiobis-(sulfosuccinimidyl-propionate) (DTSP) on the SPIMs was investigated and was proved to be able to improve adsorption quantity and stability of biomaterials. WGA was further adopted to enhance the signal taking advantage of the abundant lectin-binding sites on the bacteria surface. The immunosensor exhibited a detection limit of 102 cfu·mL−1, with a linear detection range from 102 to 107 cfu·mL−1 (r2 = 0.98). The total detection time was less than 1 h, showing its comparable sensitivity and rapid response. Furthermore, the low cost of one SPIM significantly reduced the detection cost of the biosensor. The biosensor may have great promise in food safety analysis and lead to a portable biosensing system for routine monitoring of foodborne pathogens. PMID:26251911

  17. Determination of carcinoembryonic antigen using a novel amperometric enzyme-electrode based on layer-by-layer assembly of gold nanoparticles and thionine

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Electrochemical sensing of carcinoembryonic antigen(CEA)on a gold electrode modified by the se- quential incorporation of the mediator,thionine(Thi),and gold nanoparticles(nano-Au),through co- valent linkage and electrostatic interactions onto a self-assembled monolayer configuration is de- scribed in this paper.The enzyme,horseradish peroxidase(HRP),was employed to block the possible remaining active sites of the nano-Au monolayer,avoid the non-specific adsorption,instead of bovine serum albumin(BSA),and amplify the response of the antigen-antibody reaction.Electrochemical ex- periments indicated highly efficient electron transfer by the imbedded Thi mediator and adsorbed nano-Au.The HRP kept its activity after immobilization,and the studied electrode showed sensitive response to CEA and high stability during a long period of storage.The working range for the system was 2.5 to 80.0 ng/mL with a detection limit of 0.90 ng/mL.The model membrane system in this work is a potential biosensor for mimicking the other immunosensor and enzyme sensor.

  18. Determination of carcinoembryonic antigen using a novel amperometric enzyme-electrode based on layer-by-layer assembly of gold nanoparticles and thionine

    Institute of Scientific and Technical Information of China (English)

    YUAN Ruo; ZHUO Ying; CHAI YaQin; ZHANG Ying; SUN AiLi

    2007-01-01

    Electrochemical sensing of carcinoembryonic antigen (CEA) on a gold electrode modified by the sequential incorporation of the mediator, thionine (Thi), and gold nanoparticles (nano-Au), through covalent linkage and electrostatic interactions onto a self-assembled monolayer configuration is described in this paper. The enzyme, horseradish peroxidase (HRP), was employed to block the possible remaining active sites of the nano-Au monolayer, avoid the non-specific adsorption, instead of bovine serum albumin (BSA), and amplify the response of the antigen-antibody reaction. Electrochemical experiments indicated highly efficient electron transfer by the imbedded Thi mediator and adsorbed nano-Au. The HRP kept its activity after immobilization, and the studied electrode showed sensitive response to CEA and high stability during a long period of storage. The working range for the system was 2.5 to 80.0 ng/mL with a detection limit of 0.90 ng/mL. The model membrane system in this work is a potential biosensor for mimicking the other immunosensor and enzyme sensor.

  19. Sensitive Electrochemiluminescence Immunosensor for Detection of N-Acetyl-β-d-glucosaminidase Based on a "Light-Switch" Molecule Combined with DNA Dendrimer.

    Science.gov (United States)

    Wang, Haijun; Yuan, Yali; Zhuo, Ying; Chai, Yaqin; Yuan, Ruo

    2016-06-07

    Here, a novel "light-switch" molecule of Ru (II) complex ([Ru(dcbpy)2dppz](2+)-DPEA) with self-enhanced electrochemiluminescence (ECL) property is proposed, which is almost nonemissive in aqueous solution but is brightly luminescent when it intercalates into DNA duplex. Owing to less energy loss and shorter electron-transfer distance, the intramolecular ECL reaction between the luminescent [Ru(dcbpy)2dppz](2+) and coreactive tertiary amine group in N,N-diisopropylethylenediamine (DPEA) makes the obtained "light-switch" molecule possess much higher light-switch efficiency compared with the traditional "light-switch" molecule. For increasing the loading amount and further enhancing the luminous efficiency of the "light-switch" molecule, biotin labeled DNA dendrimer (the fourth generation, G4) is prepared from Y-shape DNA by a step-by-step assembly strategy, which provides abundant intercalated sites for [Ru(dcbpy)2dppz](2+)-DPEA. Meanwhile, the obtained nanocomposite (G4-[Ru(dcbpy)2dppz](2+)-DPEA) could well bind with streptavidin labeled detection antibody (SA-Ab2) due to the existence of abundant biotin. Through sandwiched immunoreaction, an ECL immunosensor was fabricated for sensitive determination of N-acetyl-β-d-glucosaminidase (NAG), a typical biomarker for diabetic nephropathy (DN). The detemination linear range was 0.1 pg mL(-1) to 1 ng mL(-1), and the detection limit was 0.028 pg mL(-1). The developed strategy combining the ECL self-enhanced "light-switch" molecular and DNA nanotechnology offers an effective signal amplification mean and provides ample potential for further bioanalysis and clinical study.

  20. Magnetic mesoporous organic-inorganic NiCo2O4 hybrid nanomaterials for electrochemical immunosensors.

    Science.gov (United States)

    Li, Qunfang; Zeng, Lingxing; Wang, Jinchao; Tang, Dianping; Liu, Bingqian; Chen, Guonan; Wei, Mingdeng

    2011-04-01

    This study demonstrates a facile and feasible strategy toward the development of advanced electrochemical immunosensors based on chemically functionalized magnetic mesoporous organic-inorganic hybrid nanomaterials, and the preparation, characterization, and measurement of relevant properties of the immunosensor for detection of carcinoembryonic antigen (CEA, as a model analyte) in clinical immunoassays. The as-prepared nanomaterials composed of a magnetic mesoporous NiCo(2)O(4) nanosheet, an interlayer of Nafion/thionine organic molecules and a nanogold layer show good adsorption properties for the attachment of horseradish peroxidase-labeled secondary anti-CEA antibody (HRP-anti-CEA). With a sandwich-type immunoassay format, the functional bionanomaterials present good analytical properties to facilitate and modulate the way it was integrated onto the electrochemical immunosensors, and allows the detection of CEA at a concentration as low as 0.5 pg/mL. Significantly, the immunosensor could be easily regenerated by only using an external magnet without the need of any dissociated reagents. Importantly, the as-synthesized magnetic mesoporous NiCo(2)O(4) nanomaterials could be further extended for detection of other biomarkers or biocompounds.

  1. Automated-immunosensor with centrifugal fluid valves for salivary cortisol measurement

    Directory of Open Access Journals (Sweden)

    Masaki Yamaguchi

    2014-08-01

    Full Text Available Point-of-care measurement of the stress hormone cortisol will greatly facilitate the timely diagnosis and management of stress-related disorders. We describe an automated salivary cortisol immunosensor, incorporating centrifugal fluid valves and a disposable disc-chip that allows for truncated reporting of cortisol levels (<15 min. The performance characteristics of the immunosensor are optimized through select blocking agents to prevent the non-specific adsorption of proteins; immunoglobulin G (IgG polymer for the pad and milk protein for the reservoirs and the flow channels. Incorporated centrifugal fluid valves allow for rapid and repeat washings to remove impurities from the saliva samples. An optical reader and laptop computer automate the immunoassay processes and provide easily accessible digital readouts of salivary cortisol measurements. Linear regression analysis of the calibration curve for the cortisol immunosensor showed 0.92 of coefficient of multiple determination, R2, and 38.7% of coefficient of variation, CV, for a range of salivary cortisol concentrations between 0.4 and 11.3 ng/mL. The receiver operating characteristic (ROC curve analysis of human saliva samples indicate potential utility for discriminating stress disorders and underscore potential application of the biosensor in stress disorders. The performance of our salivary cortisol immunosensor approaches laboratory based tests and allows noninvasive, quantitative, and automated analysis of human salivary cortisol levels with reporting times compatible with point-of-care applications.

  2. Electrochemical impedance immunosensor for rapid detection of stressed pathogenic Staphylococcus aureus bacteria.

    Science.gov (United States)

    Bekir, Karima; Barhoumi, Houcine; Braiek, Mohamed; Chrouda, Amani; Zine, Nadia; Abid, Nabil; Maaref, Abdelrazek; Bakhrouf, Amina; Ouada, Hafedh Ben; Jaffrezic-Renault, Nicole; Mansour, Hedi Ben

    2015-10-01

    In this work, we report the adaptation of bacteria to stress conditions that induce instability of their cultural, morphological, and enzymatic characters, on which the identification of pathogenic bacteria is based. These can raise serious issues during the characterization of bacteria. The timely detection of pathogens is also a subject of great importance. For this reason, our objective is oriented towards developing an immunosensing system for rapid detection and quantification of Staphylococcus aureus. Polyclonal anti-S. aureus are immobilized onto modified gold electrode by self-assembled molecular monolayer (SAM) method. The electrochemical performances of the developed immunosensor were evaluated by impedance spectroscopy through the monitoring of the charge transfer resistance at the modified solid/liquid interface using ferri-/ferrocyanide as redox probe. The developed immunosensor was applied to detect stressed and resuscitate bacteria. As a result, a stable and reproducible immunosensor with sensitivity of 15 kΩ/decade and a detection limit of 10 CFU/mL was obtained for the S. aureus concentrations ranging from 10(1) to 10(7) CFU/mL. A low deviation in the immunosensor response (±10 %) was signed when it is exposed to stressed and not stressed bacteria.

  3. Elaboration of new method of enzyme adsorption on silicalite and nano beta zeolite for amperometric biosensor creation

    Directory of Open Access Journals (Sweden)

    Soldatkin O. O.

    2014-07-01

    Full Text Available Aim. Optimization of a new method of enzyme immobilization for amperometric biosensor creation. Methods. The amperometric biosensor with glucose oxidase immobilized on zeolites as bioselective elements and platinum disk electrode as transducers of biochemical signal into the electric one was used in the work. Results. The biosensors based on glucose oxidase adsorbed on zeolites were characterized by a higher sensitivity to glucose and a better inter-reproducibility. The best analytical characteristics were obtained for the biosensors based on nano beta zeolite. It has been found that an increase in the amount of zeolite on the surface of amperometric transducer may change such biosensor parameters as sensitivity to the substrate and duration of the analysis. Conclusions. The proposed method of enzyme immobilization by adsorption on zeolites is shown to be quite promising in the development of amperometric biosensors and therefore should be further investigated.

  4. Label-free disposable immunosensor for detection of atrazine.

    Science.gov (United States)

    Belkhamssa, Najet; Justino, Celine I L; Santos, Patrícia S M; Cardoso, Susana; Lopes, Isabel; Duarte, Armando C; Rocha-Santos, Teresa; Ksibi, Mohamed

    2016-01-01

    This work reports the construction of a fast, disposable, and label-free immunosensor for the determination of atrazine. The immunosensor is based on a field effect transistor (FET) where a network of single-walled carbon nanotubes (SWCNTs) acts as the conductor channel, constituting carbon nanotubes field effect transistors (CNTFETs). Anti-atrazine antibodies were adsorbed onto the SWCNTs and subsequently the SWCNTs were protected with Tween 20 to prevent the non-specific binding of bacteria or proteins. The principle of the immunoreaction consists in the direct adsorption of atrazine specific antibodies (anti-atrazine) to SWCNTs networks. After exposed to increasing concentrations of atrazine, the CNTFETs could be used as useful label-free platforms to detect atrazine. Under the optimal conditions, a limit of detection as low as 0.001 ng mL(-1) was obtained, which is lower than that of other methods for the atrazine detection, and in a working range between 0.001 and 10 ng mL(-1). The average recoveries obtained for real water samples spiked with atrazine varied from 87.3% to 108.0%. The results show that the constructed sensors display a high sensitivity and could be useful tools for detecting pesticides like atrazine at low concentrations. They could be also applied to the determination of atrazine in environmental aqueous samples, such as seawater and riverine water.

  5. Copper oxide assisted cysteine hierarchical structures for immunosensor application

    Energy Technology Data Exchange (ETDEWEB)

    Pandey, Chandra Mouli [Biomedical Instrumentation Section, CSIR-National Physical Laboratory, New Delhi 110012 (India); Department of Chemistry, Faculty of Science, Banaras Hindu University, Varanasi 221005 (India); Sumana, Gajjala, E-mail: sumanagajjala@gmail.com [Biomedical Instrumentation Section, CSIR-National Physical Laboratory, New Delhi 110012 (India); Tiwari, Ida [Department of Chemistry, Faculty of Science, Banaras Hindu University, Varanasi 221005 (India)

    2014-09-08

    The present work describes the promising electrochemical immunosensing strategy based on copper (II) assisted hierarchical cysteine structures (CuCys) varying from star to flower like morphology. The CuCys having average size of 10 μm have been synthesised using L-Cysteine as initial precursor in presence of copper oxide under environmentally friendly conditions in aqueous medium. To delineate the synthesis mechanism, detailed structural investigations have been carried out using characterization techniques such as X-ray diffraction, transmission electron microscopy, and Fourier transform infrared spectroscopy. The electrochemical behaviour of self-assembled CuCys on gold electrode shows surface controlled electrode reaction with an apparent electron transfer rate constant of 3.38 × 10{sup −4 }cm s{sup −1}. This innovative platform has been utilized to fabricate an immunosensor by covalently immobilizing monoclonal antibodies specific for Escherichia coli O157:H7 (E. coli). Under the optimal conditions, the fabricated immunosensor is found to be sensitive and specific for the detection of E. coli with a detection limit of 10 cfu/ml.

  6. Electrochemical immunosensor for the determination of β-casein

    Directory of Open Access Journals (Sweden)

    Judith Molinari

    2015-03-01

    Full Text Available An amperometric biosensor for the quantification of food allergens based on an inhibitory immunoassay is presented. As a proof of concept, the experimental conditions were optimized for the detection of β-casein in the 0-10 ppm range. Eight electro­che­mi­cal cells were integrated into a small-sized portable potentiostat controlled by a smart­phone via Bluetooth communication. The determination of β-casein in eight different samples can be measured with the electrochemical biosensor, which has the potential to be modified for the detection of multiple allergens.

  7. Amperometric Biosensors for Real Time Assays of Organophosphates

    Directory of Open Access Journals (Sweden)

    Kamil Kuca

    2008-09-01

    Full Text Available An amperometric biosensor based on acetylcholinesterase (AChE immobilized in gelatin was used to develop an assay for the organophosphate paraoxon. The more traditional manner employing preincubation was used for comparison between measurement procedures, although the aim of the study was to examine the performance of the biosensor for real time monitoring of organophosphates. The biosensor was immersed in a reaction chamber and paraoxon was injected inside. We were able to detect 200 pg of paraoxon within one minute or 2.5 ppb when the biosensor was preincubed in the sample solution for 15 minutes. The practical impact and expectations are discussed.

  8. Amperometric Noise at Thin Film Band Electrodes

    DEFF Research Database (Denmark)

    Larsen, Simon T.; Heien, Michael L.; Taboryski, Rafael

    2012-01-01

    Background current noise is often a significant limitation when using constant-potential amperometry for biosensor application such as amperometric recordings of transmitter release from single cells through exocytosis. In this paper, we fabricated thin-film electrodes of gold and conductive...

  9. Introduction to biosensors from electric circuits to immunosensors

    CERN Document Server

    Yoon, Jeong-Yeol

    2016-01-01

    This book equips students with a thorough understanding of various types of sensors and biosensors that can be used for chemical, biological, and biomedical applications, including but not limited to temperature sensors, strain sensor, light sensors, spectrophotometric sensors, pulse oximeter, optical fiber probes, fluorescence sensors, pH sensor, ion-selective electrodes, piezoelectric sensors, glucose sensors, DNA and immunosensors, lab-on-a-chip biosensors, paper-based lab-on-a-chip biosensors, and microcontroller-based sensors. The author treats the study of biosensors with an applications-based approach, including over 15 extensive, hands-on labs given at the end of each chapter. The material is presented using a building-block approach, beginning with the fundamentals of sensor design and temperature sensors, and ending with more complicated biosensors. New to this second edition are sections on op-amp filters, pulse oximetry, meat quality monitoring, advanced fluorescent dyes, autofluorescence, various...

  10. Capacitive immunosensor for C-reactive protein quantification

    KAUST Repository

    Sapsanis, Christos

    2015-08-02

    We report an agglutination-based immunosensor for the quantification of C-reactive protein (CRP). The developed immunoassay sensor requires approximately 15 minutes of assay time per sample and provides a sensitivity of 0.5 mg/L. We have measured the capacitance of interdigitated electrodes (IDEs) and quantified the concentration of added analyte. The proposed method is a label free detection method and hence provides rapid measurement preferable in diagnostics. We have so far been able to quantify the concentration to as low as 0.5 mg/L and as high as 10 mg/L. By quantifying CRP in serum, we can assess whether patients are prone to cardiac diseases and monitor the risk associated with such diseases. The sensor is a simple low cost structure and it can be a promising device for rapid and sensitive detection of disease markers at the point-of-care stage.

  11. Application of Ionic Liquids in Amperometric Gas Sensors.

    Science.gov (United States)

    Gębicki, Jacek; Kloskowski, Adam; Chrzanowski, Wojciech; Stepnowski, Piotr; Namiesnik, Jacek

    2016-01-01

    This article presents an analysis of available literature data on metrological parameters of the amperometric gas sensors containing ionic liquids as an electrolyte. Four mechanism types of signal generation in amperometric sensors with ionic liquid are described. Moreover, this article describes the influence of selected physico-chemical properties of the ionic liquids on the metrological parameters of these sensors. Some metrological parameters are also compared for amperometric sensors with GDE and SPE electrodes and with ionic liquids for selected analytes.

  12. Liver Cancer Detection by a Simple, Inexpensive and Effective Immunosensor with Zinc Oxide Nanoparticles

    Directory of Open Access Journals (Sweden)

    Congo Tak-Shing Ching

    2015-11-01

    Full Text Available Regular monitoring of blood α-fetoprotein (AFP and/or carcino-embryonic antigen (CEA levels is important for the routine screening of liver cancer. However, AFP and CEA have a much lower specificity than des-γ-carboxyprothrombin (DCP to detect liver cancer. Therefore, the study reported here was designed, to develop a screen-printed DCP immunosensor incorporating zinc oxide nanoparticles, for accurate determination of DCP. The designed immunosensor shows low detection limits for the detection of DCP: 0.440 ng/mL (based on impedance measurement, 0.081 ng/mL (based on real part of impedance measurement and 0.078 ng/mL (based on imaginary part of impedance measurement, within the range of 3.125 ng/mL to 2000 ng/mL. In addition, there was little interference to DCP determination by molecules such as Na+, K+, Ca2+, Cl−, glucose, urea, and uric acid. It is therefore concluded that the DCP immunosensor developed and reported here is simple, inexpensive and effective, and shows promise in the rapid screening of early-stage liver cancer at home with a point-of-care approach.

  13. Recombinant Staphylococcal protein A with cysteine residue for preparation of affinity chromatography stationary phase and immunosensor applications

    Directory of Open Access Journals (Sweden)

    Gorbatiuk O. B.

    2015-04-01

    Full Text Available Aim. Engineering of recombinant Staphylococcal protein A with cysteine residue (SPA-Cys for preparation of affinity chromatography stationary phase and formation of bioselective element of immunosensor. Methods. DNA sequences encoding IgG-binding region of SPA, His-tag and cysteine were genetically fused and expressed in E. coli. SPA-Cys was immobilized on maleimide-functionalized silica beads for affinity chromatography stationary phase preparation and on a gold sensor surface as a bioselective element of immunosensor. Results. SPA-Cys was expressed at a high-level in a soluble form. The target protein was purified and showed a high IgG-binding activity. The capacity of the obtained SPA-Cys-based affinity chromatography stationary phase was 10–12 mg of IgG /ml. The purity of eluted IgG was more than 95 % in one-step purification procedure. The developed SPA-Cys-based bioselective element of immunosensor selectively interacted with human IgG and did not interact with the control proteins. Conclusions. The recombinant Staphylococcal protein A with cysteine residue was successfully used for the preparation of affinity chromatography stationary phase and formation of the bioselective element of immunosensor.

  14. Computer-assisted electrochemical fabrication of a highly selective and sensitive amperometric nitrite sensor based on surface decoration of electrochemically reduced graphene oxide nanosheets with CoNi bimetallic alloy nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Gholivand, Mohammad-Bagher, E-mail: mbgholivand2013@gmail.com [Faculty of Chemistry, Razi University, Kermanshah 671496734 (Iran, Islamic Republic of); Jalalvand, Ali R. [Faculty of Chemistry, Razi University, Kermanshah 671496734 (Iran, Islamic Republic of); Laboratorio de Desarrollo Analítico y Quimiometría (LADAQ), Cátedra de Química Analítica I, Universidad Nacional del Litoral, Ciudad Universitaria, CC 242 (S3000ZAA), Santa Fe (Argentina); Goicoechea, Hector C. [Laboratorio de Desarrollo Analítico y Quimiometría (LADAQ), Cátedra de Química Analítica I, Universidad Nacional del Litoral, Ciudad Universitaria, CC 242 (S3000ZAA), Santa Fe (Argentina)

    2014-07-01

    For the first time, a novel, robust and very attractive statistical experimental design (ED) using minimum-run equireplicated resolution IV factorial design (Min-Run Res IV FD) coupled with face centered central composite design (FCCCD) and Derringer's desirability function (DF) was developed to fabricate a highly selective and sensitive amperometric nitrite sensor based on electrodeposition of CoNi bimetallic alloy nanoparticles (NPs) on electrochemically reduced graphene oxide (ERGO) nanosheets. The modifications were characterized by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), energy dispersive X-ray spectroscopic (EDS), scanning electron microscopy (SEM) techniques. The CoNi bimetallic alloy NPs were characterized using digital image processing (DIP) for particle counting (density estimation) and average diameter measurement. Under the identified optimal conditions, the novel sensor detects nitrite in concentration ranges of 0.1–30.0 μM and 30.0–330.0 μM with a limit of detection (LOD) of 0.05 μM. This sensor selectively detects nitrite even in the presence of high concentration of common ions and biological interferents therefore, we found that the sensor is highly selective. The sensor also demonstrated an excellent operational stability and good antifouling properties. The proposed sensor was used to the determination of nitrite in several foodstuff and water samples. - Highlights: • Eight variables were screened by Min Run Res IV FD to identify the key variables. • Mathematical models for the two studied responses were developed by FCCCD. • By using DF the responses were optimized simultaneously. • The SEM image of the modified electrode was processed by digital image processing. • The sensor was successfully applied to determination of nitrite in real samples.

  15. Microconductometric immunosensor for label-free and sensitive detection of Gram-negative bacteria.

    Science.gov (United States)

    El Ichi, Sarra; Leon, Fanny; Vossier, Ludivine; Marchandin, Helene; Errachid, Abdelhamid; Coste, Joliette; Jaffrezic-Renault, Nicole; Fournier-Wirth, Chantal

    2014-04-15

    Blood safety is a global health goal. In developed countries, bacterial contamination of platelet concentrates is the highest infectious risk in transfusion despite the current preventive strategies. We aimed to develop a conductometric biosensor for the generic, rapid and sensitive detection of Gram-negative bacteria. Our strategy is based on immunosensors: addressable magnetic nanoparticles coupled with anti-LPS antibodies were used for the generic capture of Gram-negative bacteria. Bacterial capture was characterized by impedancemetric and microscopic measurements. The results obtained with conductometric measurements allowed real-time, sensitive detection of Escherichia coli or Serratia marcescens cultures from 1 to 10(3) CFU mL(-1). The ability of the immunosensor to detect Gram negative bacteria was also tested on clinically relevant strains. The conductometric immunosensor allowed the direct detection of 10-10(3) CFU mL(-1) of Pseudomonas aeruginosa and Acinetobacter baumannii strains that were undetectable using standard immunoblot methods. Results showed that the conductometric response was not inhibited in 1% serum.

  16. A label-free electrochemical immunosensor for the detection of cardiac marker using graphene quantum dots (GQDs).

    Science.gov (United States)

    Tuteja, Satish K; Chen, Rui; Kukkar, Manil; Song, Chung Kil; Mutreja, Ruchi; Singh, Suman; Paul, Ashok K; Lee, Haiwon; Kim, Ki-Hyun; Deep, Akash; Suri, C Raman

    2016-12-15

    A label-free immunosensor based on electrochemical impedance spectroscopy has been developed for the sensitive detection of a cardiac biomarker myoglobin (cMyo). Hydrothermally synthesized graphene quantum dots (GQDs) have been used as an immobilized template on screen printed electrodes for the construction of an impedimetric sensor platform. The GQDs-modified electrode was conjugated with highly specific anti-myoglobin antibodies to develop the desired immunosensor. The values of charge transfer resistance (Rct) were monitored as a function of varying antigen concentration. The Rct value of the immunosensor showed a linear increase (from 0.20 to 0.31kΩ) in the range of 0.01-100ng/mL cMyo. The specific detection of cMyo was also made in the presence of other competing proteins. The limit of detection for the proposed immunosensor was estimated as 0.01ng/mL which is comparable to the standard ELISA techniques.

  17. Opto-Microfluidic Immunosensors: From Colorimetric to Plasmonic

    Directory of Open Access Journals (Sweden)

    Jie-Long He

    2016-02-01

    Full Text Available Optical detection has long been the most popular technique in immunosensing. Recent developments in the synthesis of luminescent probes and the fabrication of novel nanostructures enable more sensitive and efficient optical detection, which can be miniaturized and integrated with microfluidics to realize compact lab-on-a-chip immunosensors. These immunosensors are portable, economical and automated, but their sensitivity is not compromised. This review focuses on the incorporation and implementation of optical detection and microfluidics in immunosensors; it introduces the working principles of each optical detection technique and how it can be exploited in immunosensing. The recent progress in various opto-microfluidic immunosensor designs is described. Instead of being comprehensive to include all opto-microfluidic platforms, the report centers on the designs that are promising for point-of-care immunosensing diagnostics, in which ease of use, stability and cost-effective fabrication are emphasized.

  18. Electrochemical magnetic beads-based immunosensing platform for the determination of α-lactalbumin in milk.

    Science.gov (United States)

    Ruiz-Valdepeñas Montiel, Víctor; Campuzano, Susana; Torrente-Rodríguez, Rebeca M; Reviejo, A Julio; Pingarrón, José M

    2016-12-15

    Alpha-lactalbumin (α-LA) is one of the whey proteins in cows' milk that has been identified as allergenic. In this work, we present, for the first time, a very sensitive magnetic beads (MBs)-based immunosensor for the determination of α-LA. A sandwich configuration involving selective capture and horseradish peroxidase-labeled detector antibodies was implemented on carboxylic acid-modified magnetic beads, captured magnetically under the surface of a disposable screen-printed carbon electrode for amperometric detection using the hydroquinone (HQ)/H2O2 system. The α-LA immunosensor exhibited a wide linear range (37.0-5000pg/ml), a low limit of detection (LOD, 11.0pg/ml) and noteworthy selectivity against other non-target proteins. The MBs-based immunosensing platform was applied successfully for the determination of α-LA in several varieties of milk (raw and UHT cows' milk as well as human milk) and infant formulations. The results were corroborated with those obtained using a commercial ELISA method, thereby substantiating the analytical merits of this unique method.

  19. Immunosensor with Fluid Control Mechanism for Salivary Cortisol Analysis

    OpenAIRE

    Yamaguchi, Masaki; Matsuda, Yohei; Sasaki, Shohei; Sasaki, Makoto; Kadoma, Yoshihiro; Imai, Yoshikatsu; Niwa, Daisuke; Shetty, Vivek

    2012-01-01

    The purpose of this research is to demonstrate a new design for a cortisol immunosensor for the noninvasive and quantitative analysis of salivary cortisol. We propose a cortisol immunosensor with a fluid control mechanism which has both a vertical flow and a lateral flow. The detected current resulting from a competitive reaction between the sample cortisol and a glucose oxidase (GOD)-labeled cortisol conjugate was found to be inversely related to the concentration of cortisol in the sample s...

  20. An effective gold nanotubes electrode for amperometric biosensor.

    Science.gov (United States)

    Wang, Yunli; Zhu, Yingchun; Liu, Yanyan; Yang, Yu; Ruan, Qichao; Xu, Fangfang

    2010-12-01

    A sensitive and effective amperometric glucose biosensor based on gold nanotubes electrode (GNTE) was investigated. Gold nanotubes (GNTs), which were prepared by electroless plating of the metal within the pores of nanoporous polycarbonate (PC) track-etched membranes, were filled into a hollow teflon cylinder to construct a GNTE. Glucose oxidase (GOD) was immobilized on the electrode via glutaraldehyde cross-linkage method. The electrochemical properties were investigated by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The km value of the immobilized glucose oxidase on GNTE was 0.47 mM. The biosensor showed a linear range from 0.4 to 11 mM with excellent sensitivity of 8.77 microA cm(-2) mM(-1) and fast response time within 5 s.

  1. The interference of HEPES buffer during amperometric detection of ATP in clinical applications.

    Science.gov (United States)

    Masson, Jean-Francois; Gauda, Estelle; Mizaikoff, Boris; Kranz, Christine

    2008-04-01

    HEPES-based biological buffer is subject to photooxidation upon exposure to fluorescent illumination. Thereby hydrogen peroxide is generated, which interferes with amperometric oxidoreductase-based biosensors for glucose or adenosine triphosphate (ATP). These biosensors operate at an oxidation potential above 500 mV vs. the standard calomel electrode (SCE) and involve hydrogen peroxide as the electroactive molecule detected at the electrode surface. False-positive detection of ATP was observed in HEPES buffer utilizing an amperometric microbiosensor based on the co-immobilization of glucose oxidase and hexokinase for detection of ATP in biological specimens. Electrochemical, mass spectrometric, (31)P NMR, and (1)H NMR studies indicate that complexation of ATP and HEPES induced by the presence of Ca(2+) in HEPES buffer decreases the photooxidation of HEPES. Consequently, the hydrogen peroxide background concentration is reduced, thereby leading to erroneous ATP detection at the dual-enzyme microbiosensor, which determines an increase in ATP via a reduced hydrogen peroxide signal.

  2. Photocatalytic silver enhancement reaction for gravimetric immunosensors.

    Science.gov (United States)

    Seo, Hyejung; Joo, Jinmyoung; Ko, Wooree; Jung, Namchul; Jeon, Sangmin

    2010-12-17

    A novel microgravimetric immunosensor has been developed using TiO(2) nanoparticle-modified immunoassay and silver enhancement reaction. An antibody-conjugated TiO(2) nanoparticle is bound to the AFP antigen immobilized on a quartz resonator. When the nanoparticles are exposed to UV light in a silver nitrate solution, the photocatalytic reduction of silver ions results in the formation of metallic silver onto the nanoparticles and induces a decrease in the resonance frequency. The frequency change by this photocatalytic reduction reaction is three orders of magnitude larger than the change by antigen binding alone. The efficiency of the photocatalytic reaction has been found to increase with the fraction of anatase crystallites in the nanoparticles and the concentration of the AgNO(3) solution. The results highlight the potential of the photocatalytic nanoparticles for the detection of low concentrations of target molecules using gravimetric sensors.

  3. Amperometric Detection of Bacillus anthracis Spores: A Portable, Low-Cost Approach to the ELISA

    Directory of Open Access Journals (Sweden)

    Gabriel D. Peckham

    2013-01-01

    Full Text Available Antibody-based detection assays are generally robust, a desirable characteristic for in-the-field use. However, to quantify the colorimetric or fluorescent signal, these assays require expensive and fragile instruments which are ill-suited to in-the-field use. Lateral flow devices (LFDs circumvent these barriers to portability but suffer from poor sensitivity and subjective interpretation. Here, an antibody-based method for detecting Bacillus anthracis spores via amperometric signal generation is compared to ELISA and LFDs. This amperometric immunoassay uses antibody conjugated to magnetic beads and glucose oxidase (GOX along with the electron mediator 2, 6-dichlorophenolindophenol (DCPIP for production of a measurable current from a 0.4 V bias voltage. With similar sensitivity to ELISA, the assay can be completed in about 75 minutes while being completely powered and operated from a laptop computer. Immunoassay amperometry holds promise for bringing low-cost, quantitative detection of hazardous agents to the field.

  4. ZnO/36°YX-LiTaO3结构的Love波免疫传感器灵敏度研究%A study of sensitivities of Love-wave immunosensors based on ZnO/36°YX-LiTaO3 structures

    Institute of Scientific and Technical Information of China (English)

    宋明太; 周凤梅; 范理; 张淑仪

    2011-01-01

    对于ZnO/36°YX-LiTaO3结构的Love波免疫传感器,免疫传感实验的结果表明,不同的波导层厚度,免疫传感的灵敏度不同.同时,存在最佳ZnO波导层厚度,使得Love波免疫传感器的灵敏度达到最高.本文根据多层薄膜中弹性波传播理论,运用部分波求解方法,对ZnO/36.YX-LiTaO3结构的Love波器件的相速度及相对质量灵敏度与波导层的厚度的关系进行了数值计算.计算结果表明,当相对于波长归一化膜厚为0~0.1范围内,随着波导层厚度的增加,Love波器件在无负载和有负载条件下的相速度均随之减少;相对质量灵敏度则先随波导层厚度的增加而增加,继而随波导层厚度的增加而减少,即存在一个适当值使得Love波器件的相对质量灵敏度最大,该结果与实验基本吻合.%Love wave sensors have been widely used because of their high sensitivities. Meanwhile, they can be used for detecting harmful gases in gas environments and also some chemical or biological substances in liquid environments. Therefore, researches on Love wave sensors are attracting a great attention in recent years.In this paper, Love-mode immunosensors based on ZnO/36°YX- LiTaO3 structures are fabricated and the sensitivities of the immunosensors are studied. The sensitivity of Love wave sensors is affected by many factors, and the thickness of the guiding layers is a critical factor. Because the guiding layers can trap the acoustic energy to near surfaces of the Love wave sensors, the sensors display especially sensitive to surface perturbations.The experimental results show that when the thicknesses of the ZnO guiding layers are changed, the sensitivities of the immunosensors are also different; at the same time, there are optimum thicknesses of the ZnO guiding layers that make the sensitivities of the immunosensors to be the highest. Theoretical studies on Love wave devices based on ZnO/36° YX- LiTaO3 structures are carried out using partial

  5. 基于可溶性虫卵抗原的血吸虫抗体检测免疫传感器%The soluble egg antigen-based immunosensor for detection of Schistosome antibodies

    Institute of Scientific and Technical Information of China (English)

    马静; 周卓晟; 徐勇; 刘红艳; 余枫华; 刘军; 杨春秀

    2011-01-01

    目的:将免疫法特异性和电化学法灵敏性相结合,建立一种高灵敏基于可溶性虫卵抗原的电化学免疫传感器.用于血吸虫抗体的检测.方法:巯基乙胺通过吸附作用在金电极表面形成的自组装单分子层(SAM),依次滴加2.5%戊二醛和血吸虫抗原(可溶性虫卵抗原,soluble egg antigen,SEA),用以捕获抗SEA抗体(兔多抗或人多抗),然后再与辣根过氧化物酶(HRP)标记的二抗形成免疫复合物.用循环伏安法检测还原峰的电流值.结果:在底物为添加了2μl30%的双氧水及1.0 mmol/L对苯二酚的0.1 mol/L pH7.2的PBS缓冲液中,还原峰的电流值与抗SEA兔血清稀释度在10-3与10-6范围内成正比,相关系数为0.956.与抗SEA人血清稀释比在10-1与10-4范围内成正比,相关系数为0.988.在1:40稀释度条件下,血吸虫病人与正常人血清的抗SEA还原峰的电流值相差7.4μA.结论:该方法可以用于血吸虫抗体的初步检测.%Objective:Through the combination of specific immunoassay and sensitive electrochemical method, a highly sensitive electrochemical immunosensor for Schistosoma antibodies was established, which was based on the soluble egg antigen (SEA) of Schistosome.Methods: The self- assembled monolayer (SAM) was formed by mercaptoethylamine adsorbing on gold electrode surface, folowed the dropping of 2.5% glutaraldehyde and SEA to capture Schistosoma antibodies( rabbit or human polyclonal antibodies).Then the immune complex formation was combined with secondary antibodies labeled by horseradish peroxidase (HRP).The reduction peak current reduction peak current was detected by cyclic voltammetry.Results: The data showed that in the substrate of pH7.2 0.1 M PBS buffer which was amended by 1.0 mmol/L hydroquinone and the addtion of 2 μl 30% hydrogen peroxide.The reduction peak current value was proportional to the dilution of rabbit anti - schistosome serum from 10 -3 to 10 -6 and correlation coefficient is 0.956, while

  6. Immunosensor with fluid control mechanism for salivary cortisol analysis.

    Science.gov (United States)

    Yamaguchi, Masaki; Matsuda, Yohei; Sasaki, Shohei; Sasaki, Makoto; Kadoma, Yoshihiro; Imai, Yoshikatsu; Niwa, Daisuke; Shetty, Vivek

    2013-03-15

    The purpose of this research is to demonstrate a new design for a cortisol immunosensor for the noninvasive and quantitative analysis of salivary cortisol. We propose a cortisol immunosensor with a fluid control mechanism which has both a vertical flow and a lateral flow. The detected current resulting from a competitive reaction between the sample cortisol and a glucose oxidase (GOD)-labeled cortisol conjugate was found to be inversely related to the concentration of cortisol in the sample solution. A calibration curve using the relative detected current showed a R(2)=0.98 and CV=14% for a range of standard cortisol solutions corresponding to the concentrations of native salivary cortisol (0.1-10 ng/ml). The measurement could be accomplished within 35 min and the cortisol immunosensor could be reused. These results show promise for realizing an on-site and easy-to-use biosensor for cortisol. Used for evaluation of human salivary cortisol levels, the cortisol immunosensor measurement corresponded closely with commercially available ELISA method (R(2)=0.92). Our results indicate the promise of the new cortisol immunosensor for noninvasive, point of care measurement of human salivary cortisol levels.

  7. Low-interferences Determination of the Antioxidant Capacity in Fruits Juices Based on Xanthine Oxidase and Mediated Amperometric Measurements in the Reduction Mode.

    Science.gov (United States)

    Bucur, Madalina-Petruta; Radulescu, Maria-Cristina; Bucur, Bogdan; Radu, Gabriel Lucian

    2016-01-01

    A low-interferences enzymatic sensor for evaluating the antioxidant capacity was developed. Xanthine oxidase was used to produce superoxide radicals that spontaneously dismutate to hydrogen peroxide. Low xanthine concentrations were used to minimize the rapid dismutation of the superoxide radical before its fast reaction with antioxidants. The sensor operates in the reduction mode, and evaluations with low interferences of the antioxidant capacity are based on the detection of remaining hydrogen peroxide using Prussian blue electrodes at low potentials. The linear calibration graph is between 2 - 10 μM ascorbic acid. No interferences were observed from easily oxidisable substances including uric acid, which is produced in the enzymatic reaction or other substances usually found in foods. The method was used to evaluate the antioxidant capacity in different real juice samples.

  8. A choline oxidase amperometric bioassay for the detection of mustard agents based on screen-printed electrodes modified with Prussian Blue nanoparticles.

    Science.gov (United States)

    Arduini, Fabiana; Scognamiglio, Viviana; Covaia, Corrado; Amine, Aziz; Moscone, Danila; Palleschi, Giuseppe

    2015-02-13

    In this work a novel bioassay for mustard agent detection was proposed. The bioassay is based on the capability of these compounds to inhibit the enzyme choline oxidase. The enzymatic activity, which is correlated to the mustard agents, was electrochemically monitored measuring the enzymatic product, hydrogen peroxide, by means of a screen-printed electrode modified with Prussian Blue nanoparticles. Prussian Blue nanoparticles are able to electrocatalyse the hydrogen peroxide concentration reduction at low applied potential (-50 mV vs. Ag/AgCl), thus allowing the detection of the mustard agents with no electrochemical interferences. The suitability of this novel bioassay was tested with the nitrogen mustard simulant bis(2-chloroethyl)amine and the sulfur mustard simulants 2-chloroethyl ethyl sulfide and 2-chloroethyl phenyl sulfide. The bioassay proposed in this work allowed the detection of mustard agent simulants with good sensitivity and fast response, which are excellent premises for the development of a miniaturised sensor well suited for an alarm system in case of terrorist attacks.

  9. Thin-film amperometric multibiosensor for simultaneous determination of lactate and glucose in wine.

    Science.gov (United States)

    Shkotova, Lyudmyla V; Piechniakova, Nataliia Y; Kukla, Oleksandr L; Dzyadevych, Sergei V

    2016-04-15

    An amperometric multi-biosensor based on lactate and glucose oxidases has been developed for determination of lactate and glucose in wine. Gold thin-film amperometric electrodes were used as multi-transducers. Analytical characteristics of the multi-biosensor developed were studied. The minimum detectable concentration was 5×10(-6) mol/l for both glucose and lactate. High reproducibility and storage stability of the multi-biosensor are demonstrated in this paper. Lactate and glucose were determined in wine, and a good correlation was obtained with concentrations determined using high-performance liquid chromatography (correlation coefficient for glucose R(2)=0.998, for lactate R(2)=0.718).

  10. An ultra-sensitive impedimetric immunosensor for detection of the serum oncomarker CA-125 in ovarian cancer patients

    Science.gov (United States)

    Johari-Ahar, M.; Rashidi, M. R.; Barar, J.; Aghaie, M.; Mohammadnejad, D.; Ramazani, A.; Karami, P.; Coukos, G.; Omidi, Y.

    2015-02-01

    Effective treatment of ovarian cancer depends upon the early detection of the malignancy. Here, we report on the development of a new nanostructured immunosensor for early detection of cancer antigen 125 (CA-125). A gold electrode was modified with mercaptopropionic acid (MPA), and then consecutively conjugated with silica coated gold nanoparticles (AuNP@SiO2), CdSe quantum dots (QDs) and anti-CA-125 monoclonal antibody (mAb). The engineered MPA|AuNP@SiO2|QD|mAb immunosensor was characterised using transmission electron microscopy (TEM), atomic force microscopy (AFM), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Successive conjugation of AuNP@SiO2, CdSe QD and anti-CA-125 mAb onto the gold electrode resulted in sensitive detection of CA-125 with a limit of detection (LOD) of 0.0016 U mL-1 and a linear detection range (LDR) of 0-0.1 U mL-1. Based on the high sensitivity and specificity of the immunosensor, we propose this highly stable and reproducible biosensor for the early detection of CA-125.Effective treatment of ovarian cancer depends upon the early detection of the malignancy. Here, we report on the development of a new nanostructured immunosensor for early detection of cancer antigen 125 (CA-125). A gold electrode was modified with mercaptopropionic acid (MPA), and then consecutively conjugated with silica coated gold nanoparticles (AuNP@SiO2), CdSe quantum dots (QDs) and anti-CA-125 monoclonal antibody (mAb). The engineered MPA|AuNP@SiO2|QD|mAb immunosensor was characterised using transmission electron microscopy (TEM), atomic force microscopy (AFM), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Successive conjugation of AuNP@SiO2, CdSe QD and anti-CA-125 mAb onto the gold electrode resulted in sensitive detection of CA-125 with a limit of detection (LOD) of 0.0016 U mL-1 and a linear detection range (LDR) of 0-0.1 U mL-1. Based on the high sensitivity and specificity of the immunosensor, we propose

  11. Impedimetric immunosensor for human serum albumin detection on a direct aldehyde-functionalized silicon nitride surface

    Energy Technology Data Exchange (ETDEWEB)

    Caballero, David, E-mail: caballero@unistra.fr [Nanobioengineering group-IBEC, Barcelona Science Park, C/ Baldiri Reixach 10-12, 08028 Barcelona (Spain); University of Barcelona, Department of Electronics, C/ Marti i Franques 1, 08028 Barcelona (Spain); Centro de Investigacion Biomedica en Red en Bioingenieria, Biomateriales y Nanomedicina (CIBER-BBN), 50018 Zaragoza (Spain); Martinez, Elena [Nanobioengineering group-IBEC, Barcelona Science Park, C/ Baldiri Reixach 10-12, 08028 Barcelona (Spain); Centro de Investigacion Biomedica en Red en Bioingenieria, Biomateriales y Nanomedicina (CIBER-BBN), 50018 Zaragoza (Spain); Bausells, Joan [Centre Nacional de Microelectronica (CNM-IMB), CSIC, Campus UAB, 08193 Bellaterra (Spain); Errachid, Abdelhamid, E-mail: abdelhamid.errachid-el-salhi@univ-lyon1.fr [Nanobioengineering group-IBEC, Barcelona Science Park, C/ Baldiri Reixach 10-12, 08028 Barcelona (Spain); Universite Claude Bernard - Lyon 1, LSA - UMR 5180, 43 Bd du 11 novembre 1918, 69622 Villeurbanne Cedex (France); Samitier, Josep [Nanobioengineering group-IBEC, Barcelona Science Park, C/ Baldiri Reixach 10-12, 08028 Barcelona (Spain); University of Barcelona, Department of Electronics, C/ Marti i Franques 1, 08028 Barcelona (Spain); Centro de Investigacion Biomedica en Red en Bioingenieria, Biomateriales y Nanomedicina (CIBER-BBN), 50018 Zaragoza (Spain)

    2012-03-30

    Highlights: Black-Right-Pointing-Pointer An impedimetric label-free immunosensor was developed for the specific detection of human serum albumin proteins. Black-Right-Pointing-Pointer Anti-HSA antibodies were covalently immobilized on silicon nitride surfaces using a direct functionalization methodology. Black-Right-Pointing-Pointer Silicon nitride offers multiple advantages compared to other common materials. Black-Right-Pointing-Pointer The proposed sensor has high sensitivity and good selectivity for the detection of HSA proteins. - Abstract: In this work we report the fabrication and characterization of a label-free impedimetric immunosensor based on a silicon nitride (Si{sub 3}N{sub 4}) surface for the specific detection of human serum albumin (HSA) proteins. Silicon nitride provides several advantages compared with other materials commonly used, such as gold, and in particular in solid-state physics for electronic-based biosensors. However, few Si{sub 3}N{sub 4}-based biosensors have been developed; the lack of an efficient and direct protocol for the integration of biological elements with silicon-based substrates is still one of its the main drawbacks. Here, we use a direct functionalization method for the direct covalent binding of monoclonal anti-HSA antibodies on an aldehyde-functionalized Si-p/SiO{sub 2}/Si{sub 3}N{sub 4} structure. This methodology, in contrast with most of the protocols reported in literature, requires less chemical reagents, it is less time-consuming and it does not need any chemical activation. The detection capability of the immunosensor was tested by performing non-faradaic electrochemical impedance spectroscopy (EIS) measurements for the specific detection of HSA proteins. Protein concentrations within the linear range of 10{sup -13}-10{sup -7} M were detected, showing a sensitivity of 0.128 {Omega} {mu}M{sup -1} and a limit of detection of 10{sup -14} M. The specificity of the sensor was also addressed by studying the

  12. An Electrochemical Immunosensor for PBA Detection Based on Graphene-Chitosan Composite Film Modified Electrode%石墨烯-壳聚糖复合物修饰电极构建电化学免疫传感器对1-芘丁酸的检测研究

    Institute of Scientific and Technical Information of China (English)

    许双姐; 吴根英; 许贺; 柳建设; 金利通

    2012-01-01

    An highly sensitive, stable and label-free electrochemical immunosensor for the detection of 1 -pyrenebutyric acid (PBA) was developed based on graphene (GS) and chitosan (CS) composites modified glassy carbon electrode ( GS - CS/GCE ) . The proposed GS - CS/GCE was activated by the l-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride ( EDC ) and N-hydroxysuccinimide (NHS) (4:1) solution in order to covalently immobilize PAHs antibody (anti-PAHs). The morphology of the GS - CS composites film was characterized by transmission electron microscopy (TEM). In phosphate buffer solution (pH 7. 0) containing 0. 1 mol/L KC1 and 10 mmol/L K3Fe( CN)6 solution (0. 1 mol/L KC1 + 10 mmol/L K3Fe(CN)6 +0. 1 mol/L PBS(pH 7. 0)), the electrochemical behaviors of the modified electrode and the electrochemical properties of the immunosensor were studied by cyclic voltammetry ( CV) and differential pulse voltammetry ( DPV). Due to the synergistic effect of grapheme and chitosan, the peak current of GS - CS/GCE in the supporting electrolyte was largely increased, which was attributed to improve sensitivity of the immunosensor. Under the optimized conditions, the immobilization amount of antibody was significantly improved on the modified e-lectrode, resulting in amplifying the molecular recognition ability of the modified electrodes. Because of the poor conductivity of the antybody - antigen immunocomplex, the current response of the proposed immunosensor decreased linearly with increase of PBA concentration. The linear range of the immunosensor for PBA detection was 0. 1 -80 μg/L with a detection limit of 0. 03 μg/L. With excellent selectivity and reproducibility, the proposed immunosensor was applied in the determination of PBA in the real sample with spiked recoveries of 90% -105% .%采用石墨烯(GS)和壳聚糖(CS)复合膜修饰玻碳电极(GS-CS/GCE),利用1-乙基-(3-二.甲基氨基丙基)碳二亚胺盐酸盐(EDC)和N-羟基丁二酰亚胺(NHS)(4∶1)活化GS-CS

  13. Flow immunosensor detection of explosives and drugs of abuse

    Science.gov (United States)

    Kusterbeck, Anne W.; Judd, Linda L.; Yu, Hao; Myles, June; Ligler, Frances S.

    1994-03-01

    An antibody-based biosensor has been developed at the Naval Research Laboratory which is capable of detecting both drugs and explosives present at low levels in an aqueous sample. In the flow immunosensor, antibodies are immobilized onto a solid substrate, allowed to bind a fluorescently labeled signal molecule, placed in a small column and attached to a buffer flow. Upon sample introduction, an amount of the fluorescent signal molecule is released that is proportional to the concentration of applied sample. The response time of the sensor is under a minute, and multiple samples can be analyzed without the need for additional reagents. Quantitative assays are being developed for a variety of compounds, including TNT, DNT, PETN, and cocaine. The laboratory prototype has been used to study how choice of fluorophore, antibody density, and flow rate affect the signal intensity and column lifetime. A self-contained commercial instrument which can analyze up to seven different compounds from a single sample is currently being engineered under a Cooperative Research and Development Agreement.

  14. Progress of Interdigitated Array Microelectrodes Based Impedance Immunosensor%基于叉指阵列微电极的阻抗免疫传感器研究进展

    Institute of Scientific and Technical Information of China (English)

    颜小飞; 汪懋华; 安冬

    2011-01-01

    叉指阵列微电极(Interdigitated array microelectrodes,IDAM)具有检出限低、灵敏度高和信噪比好等优点,近年来在分析化学领域引起了极大的关注.阻抗免疫传感器将IDAM与免疫测定技术相结合,通过抗原抗体的特异性反应引起IDAM之间介质的阻抗变化实现对目标物的检测.本文分析了 IDAM的特点、制作材料以及电极设计参数对系统检测性能的影响,评述了IDAM阻抗免疫传感器的工作原理、等效电路分析,综述了IDAM阻抗免疫传感器在食品安全分析和临床诊断领域中的应用,并讨论了目前研究中存在的问题及其发展趋势.%Interdigitated array microclectrodes ( IDAM ) present promising advantages in terms of low detection limit, high sensitivity and increased signal-to-noise ratio, thus have received great attention in the area of analytical chemistry. According to the impedance signal induced by immune reaction of antibody and antigen on the surface of microelectrods, IDAM impedance immunosensor could be used to detect target analyte in samples specifically. This review introduced the characteristics, materials and design parameters of IDAM, discussed the principle and equivalent circuit of impedance immunosensor, and summarized their applications in food safety and clinical diagnosis. Additionally, the shortcomings of current research and developing trends in the future are also discussed.

  15. Detection of hexavalent uranium with inline and field-portable immunosensors

    Energy Technology Data Exchange (ETDEWEB)

    Melton, Scott J.; Yu, Haini; Ali, Mehnaaz F.; Williams, Kenneth H; Wilkins, Michael J.; Long, Philip E.; Blake, Diane A.

    2008-10-02

    An antibody that recognizes a chelated form of hexavalent uranium was used in the development of two different immunosensors for uranium detection. Specifically, these sensors were utilized for the analysis of groundwater samples collected during a 2007 field study of in situ bioremediation in a aquifer located at Rifle, CO. The antibody-based sensors provided data comparable to that obtained using Kinetic Phosphorescence Analysis (KPA). Thus, these novel instruments and associated reagents should provide field researchers and resource managers with valuable new tools for on-site data acquisition.

  16. Determination of phenolic compounds by a polyphenol oxidase amperometric biosensor and artificial neural network analysis.

    Science.gov (United States)

    Gutés, A; Céspedes, F; Alegret, S; del Valle, M

    2005-02-15

    The determination of phenolic compounds is significant given its toxicity, even at very low concentration levels. Amperometric determination of phenols is a simple technique available. Direct oxidation of phenols can be used, but another possibility is the use of polyphenol oxidase (tyrosinase) enzyme biosensors that oxidises the phenolic compounds into their corresponding quinones. Reduction of the resulting quinones accomplishes the amplification of the amperometric signal, as long as the result of the reduction process is the corresponding cathecol, this being able to be oxidised again by the polyphenol oxidase immobilized on the surface of the biosensor. In this communication, simultaneous determination of different phenols was carried out combining biosensor measurements with chemometric tools, in what is known as electronic tongue. The departure information used was the overlapped reduction voltammogram generated with the amperometric biosensor based on polyphenol oxidase. Artificial Neural Networks (ANN) were used for extraction and quantification of each compound. Phenol, cathecol and m-cresol formed the three-analyte study case resolved in this work. Good prediction ability was attained, and so, the separate quantification of these three phenols was accomplished.

  17. PPB级锆基安培型NO气体传感器性能的研究%Characteristics Study of the Zirconia-Based Amperometric Gas Sensors Response to the PPB-Level Concentration of NO

    Institute of Scientific and Technical Information of China (English)

    朱卫学; 金涵; 朱丽娜; 张帆; 简家文

    2015-01-01

    采用丝网印刷技术制备了以In2O3为敏感电极的锆基安培型三电极NO传感器用以探测10-9级NO气体。用X射线衍射仪(XRD)和扫描电镜(SEM)对该传感器进行了理化分析;通过测量其在不同温度和不同NO浓度的气氛中的伏安特性曲线和时间响应曲线,研究了传感器的电流输出信号和NO浓度的关系以及时间响应特性。实验表明:在350℃~500℃测试温度范围内,极化电压为-60 mV,NO浓度变化为0~900×10-9时传感器响应电流的变化值Δcurrent和NO浓度之间存在较好的线性关系并且传感器在400℃时响应值最大。在被测气体总流量为100 cm3/min时,传感器信号90%的响应和恢复时间分别为18 s和12 s。传感器信号不受CO2浓度变化的影响,传感器的响应信号在测试时间里具有较好短期稳定性,但长期稳定性有待进一步提高。本文还采用阻抗谱分析方法对传感器的响应机理进行了初步的探索。%Amperometric NO sensor based on yttria-stabilized zirconia(YSZ)with In2O3sensing electrode was pre⁃pared by screen-printing technique to detect the ppblevel of NO. The physical characteristics of the sensor were studied by the X-ray diffraction(XRD)and scanning electron microscope(SEM).The sensitivity of the sensor was studied by measuring the current-voltagecurves and response/recovery time characteristics atdifferent NO concentra⁃tions and temperatures.The results show that,inthe range of 350℃~500℃and with the polarization voltage of-60 mV,there is a good linear relationship between the variation of the current change∆currentand NO concentration , whilethe volume fraction of NO changes from 0 to 900 × 10-9.The sensor response best when the temperature is at 400℃. When the total gas flow rate is 100 cm3/min,the response and recovery time of the 90%signal of the sensors are respectively 18 s and 12 s. Sensor signal is not affected by changes in CO2 concentration

  18. Label free sensing of creatinine using a 6 GHz CMOS near-field dielectric immunosensor.

    Science.gov (United States)

    Guha, S; Warsinke, A; Tientcheu, Ch M; Schmalz, K; Meliani, C; Wenger, Ch

    2015-05-07

    In this work we present a CMOS high frequency direct immunosensor operating at 6 GHz (C-band) for label free determination of creatinine. The sensor is fabricated in standard 0.13 μm SiGe:C BiCMOS process. The report also demonstrates the ability to immobilize creatinine molecules on a Si3N4 passivation layer of the standard BiCMOS/CMOS process, therefore, evading any further need of cumbersome post processing of the fabricated sensor chip. The sensor is based on capacitive detection of the amount of non-creatinine bound antibodies binding to an immobilized creatinine layer on the passivated sensor. The chip bound antibody amount in turn corresponds indirectly to the creatinine concentration used in the incubation phase. The determination of creatinine in the concentration range of 0.88-880 μM is successfully demonstrated in this work. A sensitivity of 35 MHz/10 fold increase in creatinine concentration (during incubation) at the centre frequency of 6 GHz is gained by the immunosensor. The results are compared with a standard optical measurement technique and the dynamic range and sensitivity is of the order of the established optical indication technique. The C-band immunosensor chip comprising an area of 0.3 mm(2) reduces the sensing area considerably, therefore, requiring a sample volume as low as 2 μl. The small analyte sample volume and label free approach also reduce the experimental costs in addition to the low fabrication costs offered by the batch fabrication technique of CMOS/BiCMOS process.

  19. A self-amplified transistor immunosensor under dual gate operation: highly sensitive detection of hepatitis B surface antigen.

    Science.gov (United States)

    Lee, I-K; Jeun, M; Jang, H-J; Cho, W-J; Lee, K H

    2015-10-28

    Ion-sensitive field-effect transistors (ISFETs), although they have attracted considerable attention as effective immunosensors, have still not been adopted for practical applications owing to several problems: (1) the poor sensitivity caused by the short Debye screening length in media with high ion concentration, (2) time-consuming preconditioning processes for achieving the highly-diluted media, and (3) the low durability caused by undesirable ions such as sodium chloride in the media. Here, we propose a highly sensitive immunosensor based on a self-amplified transistor under dual gate operation (immuno-DG ISFET) for the detection of hepatitis B surface antigen. To address the challenges in current ISFET-based immunosensors, we have enhanced the sensitivity of an immunosensor by precisely tailoring the nanostructure of the transistor. In the pH sensing test, the immuno-DG ISFET showed superior sensitivity (2085.53 mV per pH) to both standard ISFET under single gate operation (58.88 mV per pH) and DG ISFET with a non-tailored transistor (381.14 mV per pH). Moreover, concerning the detection of hepatitis B surface antigens (HBsAg) using the immuno-DG ISFET, we have successfully detected trace amounts of HBsAg (22.5 fg mL(-1)) in a non-diluted 1× PBS medium with a high sensitivity of 690 mV. Our results demonstrate that the proposed immuno-DG ISFET can be a biosensor platform for practical use in the diagnosis of various diseases.

  20. Electrochemical impedance spectroscopy for analytical determination of paraquat in meconium samples using an immunosensor modified with fullerene, ferrocene and ionic liquid

    Energy Technology Data Exchange (ETDEWEB)

    Sun Xiulan [State Key Laboratory of Food Science and Technology, Wuxi 214122 (China); Li Zaijun, E-mail: zaijunli@263.ne [School of Chemical and Materials Engineering, Jiangnan University, Lihu Road 1800, Wuxi 214122 (China); Cai, Yan; Wei, Zhilei [School of Chemical and Materials Engineering, Jiangnan University, Lihu Road 1800, Wuxi 214122 (China); Fang Yinjun; Ren Guoxiao; Huang Yaru [Zhejiang Zanyu Technology Limited Corporation, Hangzhou 311215 (China)

    2011-01-01

    The paper reports a highly sensitive electrochemical immunosensor for the detection of paraquat. The immunosensor bases on glassy carbon electrode modified with a composite made from fullerene, ferrocene and the ionic liquid. The components were immobilized on the electrode surface by chitosan. The antibody of paraquat was covalently conjugated to the surface which was then blocked with bovine serum albumin. Analytical characteristics of the immunosensor were investigated by electrochemical impedance spectroscopy. It offers good repeatability (RSD = 1.5%), a stability of more than 150 days, an impedimetric response to paraquat in the range from 3.89 x 10{sup -11} to 4.0 x 10{sup -8} mol L{sup -1}, and a detection limit (S/N = 3) of 9.0 x 10{sup -12} mol L{sup -1}. The effects of omitting fullerene and the ionic liquid were well tested. The results indicated that sensitivity of the immunosensor is 3.7-fold better if fullerene and ionic liquid are used. This demonstrates that fullerene facilitates electron transfer on surface of the electrode due to unique electrochemical properties, while the ionic liquid provides biocompatible microenvironment for the antibody, which results in the enhanced sensitivity and stability. Moreover, surface morphology feature and electrochemical properties of the electrode were also examined. The method was satisfactorily applied to the determination of paraquat in meconium.

  1. Amperometric electrochemical microsystem for a miniaturized protein biosensor array.

    Science.gov (United States)

    Chao Yang; Yue Huang; Hassler, B L; Worden, R M; Mason, A J

    2009-06-01

    Protein-based bioelectrochemical interfaces offer great potential for rapid detection, continuous use, and miniaturized sensor arrays. This paper introduces a microsystem platform that enables multiple bioelectrochemical interfaces to be interrogated simultaneously by an onchip amperometric readout system. A post-complementary metal-oxide semiconductor (CMOS) fabrication procedure is described that permits the formation of planar electrode arrays and self assembly of biosensor interfaces on the electrodes. The onchip, 0.5-mum CMOS readout electronics include a compact potentiostat that supports a very broad range of input currents-6 pA to 10 muA-to accommodate diverse biosensor interfaces. The 2.3 times 2.2-mm chip operates from a 5-V supply at 0.6 mA. A prototype electrochemical sensor platform, including an onchip potentiostat and miniaturized biosensor array, was characterized by using cyclic voltammetry. The linear relationship between the oxidization peak values and the concentrations of target analytes in the solution verifies functionality of the system and demonstrates the potential for future implementations of this platform in high sensitivity, low cost, and onchip protein-based sensor arrays.

  2. Amperometric Bioelectronic Tongue for glucose determination

    Directory of Open Access Journals (Sweden)

    Yazan Al-Issa

    2015-03-01

    Full Text Available An amperometric Bioelectronic Tongue is reported for glucose determination that contains eight sensor electrodes constructed using different metal electrodes (Pt, Au, oxidoreductase enzymes (glucose oxidase, ascorbate oxidase, uricase, and membrane coatings (Nafion, chitosan. The response to varying concentrations of glucose, ascorbic acid, uric acid, and acetaminophen was tested for two models, concentration determination by current density measurements at individual electrodes and concentration determination by a linear regression model for the entire electrode array. The reduced chi-squared for the full array model was found to be about one order of magnitude lower than that for the individual-electrode model. Discrimination of glucose from chemical interference by the other three species is accomplished through a combination of enzyme catalysis, metal electrocatalysis, and membrane surface charge. The benefit of incorporating enzyme electrodes into the sensor array is illustrated by the lower correlation coefficients between different enzyme electrodes relative to non-enzyme coated electrodes. This approach can be more generally applied to detection of other substrates of oxidoreductase enzymes.

  3. Optimization of bioselective membrane of amperometric enzyme sensor on basis of glucose oxidase using NH2-modified multi-wall carbone nanotubes

    Directory of Open Access Journals (Sweden)

    Korpan Ya. I.

    2010-02-01

    Full Text Available Aim. To investigate a possibility of application of multi-wall carbone nanotubes modified with NH2-groups (MWCNT-NH2 for creation of sensitive elements of the amperometric biosensor based on immobilized oxidoreductases, in particular, glucose oxidase (GOD. To study electrochemical properties of the membranes obtained. Methods. Experiments were carried out with amperometric methods using the ìStat 200 device («DropSens», Spain. The enzymes were immobilised in glutaraldehyde vapour. Results. The method of formation of bioselective matrix based on immobilised GOD with MNP-NH2 on the surface of gold amperometric electrodes was optimised. Optimal working conditions of the biosensor developed were determined. Conclusion. MWCNT integration into a bioselective matrix improves the biosensor analytical characteristics which means: higher signal value, wider linear range of glucose analysis, and possibility of substrate determination in wide range of working potential.

  4. Novel electrode systems for amperometric sensing: the case of titanium

    Science.gov (United States)

    Terzi, F.; Pigani, L.; Zanardi, C.; Zanfrognini, B.; Ruggeri, S.; Maccaferri, G.; Seeber, R.

    2014-10-01

    After working for years on organic materials, e.g., polythiophenes and relevant composites with metal nanoparticles, we shifted our attention to unusual metals, chosen as candidates to effective amperometric sensing on the basis of the atomic structure and crystalline properties. The present contribution aims at proposing an electrode material rarely employed in electroanalysis, namely Ti. We have experimented that the peculiar nature of Ti leads to electrochemical behavior quite different with respect to the conventional electrode materials, including those based on TiO2 (nano)particles. Our work focuses on the determination of strong oxidizing species, namely H2O2 and HClO, and noble metal ions, namely Au(III). Strong oxidizing species are commodity chemicals employed in a number of different industrial processes, in which usually high concentration levels should be monitored. The procedures proposed have been successfully applied also in complex matrices, such as detergent samples. As to Au(III) determination, it also constitutes a crucial tool in order to increase the efficiency of hydrometallurgic processes and of the recovery of precious materials from electronic waste. Ti electrodes allow the determination of dissolved Au species in the presence of other metal ions. In any cases the electrodes exhibit reproducible and repeatable electrochemical responses, even in the presence of high concentration of organic fouling species typical of bio-sorption processes.

  5. Prediction of wastewater quality using amperometric bioelectronic tongues.

    Science.gov (United States)

    Czolkos, Ilja; Dock, Eva; Tønning, Erik; Christensen, Jakob; Winther-Nielsen, Margrethe; Carlsson, Charlotte; Mojzíková, Renata; Skládal, Petr; Wollenberger, Ulla; Nørgaard, Lars; Ruzgas, Tautgirdas; Emnéus, Jenny

    2016-01-15

    Wastewater samples from a Swedish chemi-thermo-mechanical pulp (CTMP) mill collected at different purification stages in a wastewater treatment plant (WWTP) were analyzed with an amperometric enzyme-based biosensor array in a flow-injection system. In order to resolve the complex composition of the wastewater, the array consists of several sensing elements which yield a multidimensional response. We used principal component analysis (PCA) to decompose the array's responses, and found that wastewater with different degrees of pollution can be differentiated. With the help of partial least squares regression (PLS-R), we could link the sensor responses to the Microtox® toxicity parameter, as well as to global organic pollution parameters (COD, BOD, and TOC). From investigating the influences of individual sensors in the array, it was found that the best models were in most cases obtained when all sensors in the array were included in the PLS-R model. We find that fast simultaneous determination of several global environmental parameters characterizing wastewaters is possible with this kind of biosensor array, in particular because of the link between the sensor responses and the biological effect onto the ecosystem into which the wastewater would be released. In conjunction with multivariate data analysis tools, there is strong potential to reduce the total time until a result is yielded from days to a few minutes.

  6. Microfluidic immunosensor for rapid and highly-sensitive salivary cortisol quantification.

    Science.gov (United States)

    Pinto, V; Sousa, P; Catarino, S O; Correia-Neves, M; Minas, G

    2017-04-15

    This paper presents a novel poly(dimethylsiloxane) (PDMS) microfluidic immunosensor that integrates a complementary metal-oxide-semiconductor (CMOS) optical detection system for a rapid and highly-sensitive quantification of salivary cortisol. The simple and non-invasive method of saliva sampling provides an interesting alternative to the blood, allowing a fast sampling at short intervals, relevant for many clinical diagnostic applications. The developed approach is based on the covalent immobilization of a coating antibody (Ab), a polyclonal anti-IgG, onto a treated PDMS surface. The coating Ab binds the capture Ab, an IgG specific for cortisol, allowing its correct orientation. Horseradish peroxidase (HRP)-labelled cortisol is added to compete with the cortisol in the sample, for the capture Ab binding sites. The HRP-labelled cortisol, bonded to the capture Ab, is measured through the HRP enzyme and the tetramethylbenzidine (TMB) substrate reaction. The cortisol quantification is performed by colorimetric detection of HRP-labelled cortisol, through optical absorption at 450nm, using a CMOS silicon photodiode as the photodetector. Under the developed optimized conditions presented here, e.g., microfluidic channels geometry, immobilization method and immunoassay conditions, the immunosensor shows a linear range of detection between 0.01-20ng/mL, a limit of detection (LOD) of 18pg/mL and an analysis time of 35min, featuring a great potential for point-of-care applications requiring continuous monitoring of the salivary cortisol levels during a circadian cycle.

  7. An impedimetric immunosensor for the detection of autoantibodies directed against gliadins.

    Science.gov (United States)

    Balkenhohl, T; Lisdat, F

    2007-04-01

    An immunosensor has been developed for the detection of autoantibodies directed against wheat gliadin, a protein fraction of cereal gluten which is involved in celiac disease. The immunosensor is based on the immobilization of gliadins onto gold electrodes covered with a polyelectrolyte layer of poly(4-styrenesulfonic acid sodium salt). The immobilization was monitored by quartz crystal microbalance (QCM) analysis. The antigen-antibody interaction signal was amplified by an incubation step with peroxidase-labeled immunoglobulins and subsequent peroxidase-catalyzed oxidation of 3-amino-9-ethylcarbazole (AEC). Changes in the insulating properties of the electrode layer were measured by electrochemical impedance spectroscopy (EIS) in the presence of ferri/ferro-cyanide. Impedance spectra could be fitted to a Randles equivalent circuit with high accuracy. Exposing the sensor electrodes to various antigliadin antibody concentrations resulted in proportional changes in the charge transfer resistance. A calibration graph for the detection of antigliadin antibodies was established for antibody concentrations between 10(-8) and 10(-6) M. Finally, the sensor was used for the determination of antigliadin autoantibodies of the IgG and IgA type in several human sera.

  8. Electrochemical Immunosensor Based on AuNPs/PDDA-GO for Detection of SirT1%基于AuNPs/PDDA-GO纳米复合物的电化学免疫传感器的构建及对SirT1蛋白的检测

    Institute of Scientific and Technical Information of China (English)

    毕文姬; 陆丹琴; 符莹; 黄齐林; 徐志爱; 张文

    2013-01-01

    基于AuNPs/PDDA-GO纳米复合物制备了一种新型电化学免疫传感器,并将其用于SirT1的检测.首先,在电极表面修饰复合材料AuNPs/PDDA-GO,然后将目标蛋白SirT1固定到修饰了AuNPs/PDDA-GO的电极表面,再通过特异性免疫反应结合一抗(Ab1)和辣根过氧化酶标记的二抗分子(HRP-Ab2),最后用示差脉冲伏安法检测电流信号,实现了对SirT1蛋白水平的测定.在优化的实验条件下,SirT1蛋白的浓度在0.1~100 ng/mL范围内与响应电流呈良好线性关系,检出限为0.029 ng/mL.%AuNPs/PDDA-GO nanocomposite was produced by combining poly(diallyldimethylammonium chloride) (PDDA) functionalized graphene oxide nanosheets (PDDA-GO) and gold nanoparticles (AuNPs)through self-assembly method,which offered an efficient platform for antibody immobilization with good biocompatibility.A new type of electrochemical immunosensor based on AuNPs/PDDA-GO nanocomposite for the detection of SirT1 was then developed.The immunosensor construction consisted of the immobilization of the hybrid architecture,the immunoreaction of target antigen SirT1 and capture antibody(Ab1),followed by the binding of HRP-Ab2.The designed immunoassay showed an excellent analytical performance,and exhibited a wide dynamic response range for SirT1 from 0.1 ng/mL to 100 ng/mL with a detection limit of 0.029 ng/mL.Overall,this developed strategy is proved as a simple,cost-effective method,and can be easily extended to other protein analysis.

  9. 基于纳米酞菁钴修饰的石墨烯作为过氧化物模拟酶的信号放大型免疫传感器的研究%A signal amplification immunosensor based on nano-cobalt phthalocyanine modified graphene as a peroxidase simulated enzyme

    Institute of Scientific and Technical Information of China (English)

    杨哲涵; 汪娟; 卓颖; 柴雅琴

    2015-01-01

    In this work, a signal-amplified electrochemical immunosensor based on cobalt phthalocyanine nanoparticles decorated graphene oxide nanocomposite (NanoCoPc/GO) was proposed for sensitive detection procalcitonin (PCT). Here, both cobalt phthalocyanine nanoparticles and graphene oxide exhibited an intrinsic enzyme mimetic activity similar to natural peroxidase. In the present of H2O2, an enhanced electrochemical signal was obtained as the catalysis of NanoCoPc/GO to H2O2. Importantly, NanoCoPc/GO as peroxidase mimetic applied to electrochemical amplification system could avoid the disadvantage of nature enzymes such as expensive and easily denatured by environmental changes. The proposed immunosensor showed good sensitivity for quantitative determination of PCT.%该文利用酞菁钴纳米粒子修饰氧化石墨烯(NanoCoPc/GO)用于构建无酶的信号放大型电化学免疫传感器来灵敏地检测降钙素原(PCT)。纳米酞菁钴和氧化石墨烯都具有类似于天然过氧化物酶的性质可以催化氧化H2O2。因此,当H2O2的存在时,NanoCoPc/GO通过催化H2O2实现对电活性物质的信号的放大。NanoCoPc/GO作为模拟酶用于电化学放大时,可以避免天然酶的缺点比如价格昂贵和容易随着环境变化而发生变性。结果表明,该免疫传感器检测PCT的线性范围在0.025~5.0 ng/mL,最低检测限为8 pg/mL。

  10. Studies of Active Ingredients in Cough Syrup by Capillary Zone Electrophoresis with Amperometric Detection

    Institute of Scientific and Technical Information of China (English)

    ZHOU Tian-shu; WANG Ai-fang; WU Fang; SHI Guo-yue; FANG Yu-zhi

    2003-01-01

    The present paper covers a simple, reliable and reproducible method, based on capillary zone electrophoresis(CZE) with amperometric detection(AD), for the separation and the determination of ephedrine hydrochloride, promethazine hydrochloride and codeine phosphate. Under the optimal conditions, the three analytes were base-line separated completely within 16 min. Good linear relationships between the peak heights and the concentrations of the three analytes were obtained with the correlation coefficients better than 0.9993. The method was directly applied to the determination of the active ingredients in pharmaceutical preparations and the assay results were satisfactory.

  11. A novel photoelectrochemical immunosensor by integration of nanobody and TiO{sub 2} nanotubes for sensitive detection of serum cystatin C

    Energy Technology Data Exchange (ETDEWEB)

    Mi, Li [School of Chemistry and Chemical Engineering, Southeast University, Nanjing 211189 (China); Wang, Pingyan; Yan, Junrong [Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203 (China); Qian, Jing; Lu, Jusheng; Yu, Jiachao [School of Chemistry and Chemical Engineering, Southeast University, Nanjing 211189 (China); Wang, Yuzhen; Liu, Hong [Plexera LLC, WA 98072 (United States); Zhu, Min [Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203 (China); Wan, Yakun, E-mail: ywansystemsbiology@gmail.com [Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203 (China); Jiangsu Nanobody Engineering and Research Center, Nantong 226010 (China); Liu, Songqin, E-mail: liusq@seu.edu.cn [School of Chemistry and Chemical Engineering, Southeast University, Nanjing 211189 (China)

    2016-01-01

    Cystatin C (CysC) is a sensitive marker for the estimation of the glomerular filtration rate and the clinical diagnosis of different diseases. In this paper, CysC-specific nanobodies (Nbs) were isolated from a phage display nanobody library. A simple and sensitive photoelectrochemical immunosensor based on TiO{sub 2} nanotube arrays (TNAs) was proposed for the sensitive detection of CysC. The TiO{sub 2} nanotube arrays deposited by electrochemical anodization displayed a high and stable photocurrent response under irradiation. After coupling CysC-specific nanobody to TNA (Nb/TNA), the proposed immunosensor for CysC can be utilized for tracking the photocurrent change of Nb/TNA caused by immunoreactions between CysC and the immobilized CysC-specific Nb. This allowed for the determination of CysC with a calibration range from 0.72 pM to 7.19 nM. The variation of the photocurrent was in a linear relationship with the logarithm of the CysC concentration in the range of 0.72 pM–3.6 nM. The immunosensor had a correlation coefficient of 0.97 and a detection limit of 0.14 pM at a signal-to-noise ratio of 3. The proposed immunosensor showed satisfactory intra- and inter-assay accuracy, high selectivity and good stability. As a result, this proposed strategy would offer a novel and simple approach for the detection of immunoreactions, provide new insights in popularizing the diagnosis of CysC, and extend the application of TiO{sub 2} nanotubes. - Highlights: • CysC-specific nanobody to CysC is isolated from phage display nanobody library. • A photoelectrochemical immunosensor for CysC develops by Nb modified TNA. • An excellent sensitivity and good selectivity of CysC sensing was obtained.

  12. Development and characterization of a microfluidic glucose sensing system based on an enzymatic microreactor and chemiluminescence detection

    NARCIS (Netherlands)

    Moon, B. -U; de Vries, M. G.; Westerink, B. H. C.; Verpoorte, E.

    2012-01-01

    Chemiluminescence detection was developed as an alternative to amperometric detection for glucose analysis in a portable, microfluidics-based continuous glucose monitoring system. Amperometric detection allows easy determination of hydrogen peroxide, a product of the glucose oxidase-catalyzed reacti

  13. Electrochemical immunosensor for ultrasensitive detection of microcystin-LR based on graphene-gold nanocomposite/functional conducting polymer/gold nanoparticle/ionic liquid composite film with electrodeposition.

    Science.gov (United States)

    Ruiyi, Li; Qianfang, Xia; Zaijun, Li; Xiulan, Sun; Junkang, Liu

    2013-06-15

    The study developed an electrochemical immunosensor for ultrasensitive detection of microcystin-LR in water. Graphene oxide and chloroauric acid were alternately electrodeposited on the surface of glassy carbon electrode for 20 cycles to fabricate graphene-gold nanocomposite. The composite was characterized and its apparent heterogeneous electron transfer rate constant (37.28±0.16 cm s (-1)) was estimated by Laviron's model. To immobilize microcystin-LR antibody and improve the electrical conductivity, 2,5-di-(2-thienyl)-1-pyrrole-1-(p-benzoic acid) and chloroauric acid were electrodeposited on the modified electrode in sequence. The ionic liquid was then dropped on the electrode surface and finally microcystin-LR antibody was covalently connected to the conducting polymer film. Experiment showed the electrochemical technique offers control over reaction parameters and excellent repeatability. The graphene-gold nanocomposite and gold nanoparticles enhance electron transfer of Fe(CN)6(3-/4-) to the electrode. The ionic liquid, 1-isobutyl-3-methylimidazolium bis(trifluoromethane-sulfonyl)imide, improves stability of the antibody. The sensor displays good repeatability (RSD=1.2%), sensitive electrochemical response to microcystin-LR in the range of 1.0×10(-16)-8.0×10(-15)M and detection limit of 3.7×10(-17)M (S/N=3). The peak current change of the sensor after and before incubation with 2.0×10(-15)M of microcystin-LR can retain 95% over a 20-weeks storage period. Proposed method presents remarkable improvement of sensitivity, repeatability and stability when compared to present microcystin-LR sensors. It has been successfully applied to the microcystin-LR determination in water samples with a spiked recovery in the range of 96.3-105.8%.

  14. An Ultrasensitive Electrochemical Immunosensor for HIV p24 Based on Fe3O4@SiO2 Nanomagnetic Probes and Nanogold Colloid-Labeled Enzyme–Antibody Copolymer as Signal Tag

    Directory of Open Access Journals (Sweden)

    Tianhua Li

    2013-03-01

    Full Text Available An ultrasensitive portable electrochemical immunosensor for human immunodeficiency virus p24 (HIV p24 antigen detection has been developed, whereby the detection sensitivity was 1000 times higher than that of the ELISA method. Firstly, a novel HRP enzyme–antibody copolymer (EV-p24 Ab2 was synthesized through an EnVision regent (EV, a dextrin amine skeleton anchoring more than 100 molecules of HRP and 15 molecules of anti IgG, then incubated in the secondary antibody of p24. Secondly, the copolymer was immobilized on the gold nanocolloids (AuNPs to fabricate a novel signal tag (AuNPs/EV-p24 Ab2. Subsequently, a sandwich-type immunoreaction would take place between the capture probe (silicon dioxide-coated magnetic Fe3O4 nanoparticles (MNPs labeled with the primary p24 antibody (MNPs-p24 Ab1, p24 (different concentrations and the signal tag [AuNPs/EV-p24 Ab2] to form the immunocomplex. Finally, the immunocomplex was absorbed on the surface of screen printed carbon electrode (SPCE by a magnet and immersed in the o-hydroxyl phenol (HQ and H2O2. The large amounts of HRP on the signal tag can catalyze the oxidation of HQ by H2O2, which can induce an amplified reductive current. Moreover, the capture probe could improve the accumulation ability of p24 and facilitate its separation from the substrate through the magnet. Under optimal conditions, the proposed immunoassay exhibited good sensitivity to p24 within a certain concentration range from 0.001 to 10.00 ng/mL, with a detection limit of 0.5 pg/mL (S/N = 3. The proposed method can be used for real-time and early detection of HIV-infected people.

  15. Determination of cyanide in microsamples by means of capillary flow injection analysis with amperometric detection

    Energy Technology Data Exchange (ETDEWEB)

    Backofen, U. [Leipzig Univ. (Germany). Sektion 8 - Chemie; Matysik, F.-M. [Department of Chemistry, University of Coimbra (Portugal); Werner, G. [Leipzig Univ. (Germany). Sektion 8 - Chemie

    1996-10-01

    A new approach for determining cyanide in microsamples is described. The method is based on capillary flow injection analysis (CFIA) with amperometric detection. The sensing electrode is a silver-plated microdisk electrode, where cyanide can react under formation of a dicyanoargentate complex. A remarkably low mass detection limit of 231 fmol cyanide is obtained for an injection volume of 60 nl. The sample throughput of the CFIA-arrangement is comparable with a conventional sized FIA-system. A practical application is given by analyzing the cyanide (amygdalin) concentration in apple kernels. (orig.). With 5 figs.

  16. Amperometric biosensor for Salmonella typhimurium detection in milk

    Science.gov (United States)

    This paper reports an amperometric biosensor for rapid and sensitive Salmonella Typhimurium detection in milk. The biosensor was assembled from the self-assembled monolayers technique on a gold surface. In this device, polyclonal antibodies were oriented by protein A. The biosensor structure was cha...

  17. 0.3 MM Diameter Flexible Amperometric Lactate Probe.

    Science.gov (United States)

    1993-01-26

    also the oxidation of the FADH 2 by 02, whereby H202 is formed. LOX-FAD + Lactate -, Pvruvate + LOX-FADH2 (1) Amperometric lactate sensors have been...tile reduced forms of a diffusional mediator, such :is 0- H-,O, or Fc-/Fc (where Fc is a ferrocene derivativei. 5 -10 FAD/ FADH - centers of LOX were also

  18. Highly sensitive potentiometric immunosensor for hepatitis B surface antigen diagnosis

    Institute of Scientific and Technical Information of China (English)

    YUAN; Ruo; TANG; Dianping; CHAI; Yaqin; ZHANG; Lingyan; LI

    2005-01-01

    A highly sensitive potentiometric immunosensor for the diagnoses of epidemic diseases has been developed by means of self-assembly to immobilize hepatitis B surface antibody (HBsAb) for the detection of hepatitis B surface antigen (HBsAg) as a model. At first, the Nafion containing -SO3- groups was immobilized on a platinum electrode surface to absorb the -NH3+ groups of antibody molecules via the opposite-charged adsorption technique, in the meantime, hepatitis B surface antibodies were adsorbed onto the surface of Au nanoparticles, then hepatitis B surface antibodies and Au nanopartilces were entrapped into polyvinyl butyral on the surface of Nafion film. The modified procedure was further characterized by electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). The influence and factors influencing the performance of resulting immunosensor were studied in detail. The resulting immunosensor exhibited sigmoid curve with log HBsAg concentrations, high sensitivity, wide linear range from 26 to 1280 ng·mL-1 with a detection limit of 3.1 ng·mL-1, rapid potentiometric response (4 months). Analytical results of clinical samples show that the developed immunoassay is comparable with the enzyme-linked immunosorbent assays (ELISAs) method, implying a promising alternative approach for detecting HBsAg in the clinical diagnosis.

  19. Bioethanol in Biofuels Checked by an Amperometric Organic Phase Enzyme Electrode (OPEE Working in “Substrate Antagonism” Format

    Directory of Open Access Journals (Sweden)

    Mauro Tomassetti

    2016-08-01

    Full Text Available The bioethanol content of two samples of biofuels was determined directly, after simple dilution in decane, by means of an amperometric catalase enzyme biosensor working in the organic phase, based on substrate antagonisms format. The results were good from the point of view of accuracy, and satisfactory for what concerns the recovery test by the standard addition method. Limit of detection (LOD was on the order of 2.5 × 10−5 M.

  20. Automatic bionalyzer using an integrated amperometric biosensor for the determination of L-malic acid in wines.

    Science.gov (United States)

    Vargas, E; Ruiz, M A; Ferrero, F J; Campuzano, S; Ruiz-Valdepeñas Montiel, V; Reviejo, A J; Pingarrón, J M

    2016-09-01

    A new automatic bioanalyzer for L-malic acid using an integrated amperometric biosensor as detector is reported for the first time in this work. The biosensor is constructed by gold film sputtering deposition on a stainless steel disk electrode and co-immobilization of the enzymes malate dehydrogenase (MDH) and diaphorase (DP) together with the redox mediator tetrathiafulvalene (TTF) by means of dialysis membrane. The analytical performance of the biosensor was evaluated when it was used as amperometric detector in three different analytical methodologies: stirred solutions, semiautomatic FIA system and automatic bioanalyzer. The bienzyme biosensor exhibited great analytical performance in terms of sensitivity, selectivity and reproducibility of the measurements and its usefulness was demonstrated by analyzing wine reference materials with certified content of L-malic acid. The attractive analytical and operational characteristics demonstrated by the automatic bioanalyzer make it a promising simple, rapid and field-based tool for routine wine and fruit control.

  1. Current Fluctuation Measurements of Amperometric Gas Sensors Constructed with Three Different Technology Procedures

    Directory of Open Access Journals (Sweden)

    Sedlak Petr

    2016-12-01

    Full Text Available Electrochemical amperometric gas sensors represent a well-established and versatile type of devices with unique features: good sensitivity and stability, short response/recovery times, and low power consumption. These sensors operate at room temperature, and therefore have been applied in monitoring air pollutants and detection of toxic and hazardous gases in a number of areas. Some drawbacks of classical electrochemical sensors are overcome by the solid polymer electrolyte (SPE based on ionic liquids. This work presents evaluation of an SPE-based amperometric sensor from the point of view of current fluctuations. The sensor is based on a novel three-electrode sensor platform with solid polymer electrolytes containing ionic liquid for detection of nitrogen dioxide − a highly toxic gas that is harmful to the environment and presenting a possible threat to human health even at low concentrations. The paper focuses on using noise measurement (electric current fluctuation measurement for evaluation of electrochemical sensors which were constructed by different fabrication processes: (i lift-off and drop-casting technology, (ii screen printing technology on a ceramic substrate and (iii screen printing on a flexible substrate.

  2. Thermally reduced graphene oxide: The study and use for reagentless amperometric D-fructose biosensors.

    Science.gov (United States)

    Šakinytė, Ieva; Barkauskas, Jurgis; Gaidukevič, Justina; Razumienė, Julija

    2015-11-01

    Aiming to create reagentless amperometric D-fructose biosensor, graphene based electrode materials have been synthesized by newly proposed thermal reduction of graphene oxide. The method allowed to separate and collect different fractions of thermally reduced graphene oxide (TRGO) with different physicochemical properties. The structural characteristics and surface morphologies of TRGO fractions were evaluated using SEM, XRD, TGA analysis, Raman spectroscopy and BET measurements. Three different fractions of TRGO were tested as electrode materials for D-fructose amperometric biosensors. The direct electron transfer (DET) from the active site of D-fructose dehydrogenase (FDH) to the electrode was achieved with all TRGO fractions. High values of the sensitivity (up to 14.5 μA mM(-1) cm(-2)) are of the same order as these for other D-fructose sensors based on the synergistic mediated processes. The relationships between the structure of TRGO fractions and the molecular processes determining the effect of DET in bioelectrocatalysis by FDH have been studied. Stability of the D-fructose biosensors was also assessed. The best results were achieved when immobilization of FDH was performed using a crosslinking with glutaraldehyde. For the best group, after a period of 5 days the sensitivity of the biosensor for D-fructose determination decreased by less than 20%.

  3. Iridium Oxide Film-Enhanced Impedance Immunosensor for Rapid Detection of Carcinoembyronic Antigen

    Institute of Scientific and Technical Information of China (English)

    DING,Yan-Jun; WANG,Hua; JIANG,Jian-Hui; SHEN,Guo-Li; YU,Ru-Qin

    2007-01-01

    A simple, rapid and sensitive impedance immunosensor based on iridium oxide (IrOx) thin film for the detection of carcinoembyronic antigen (CEA) in human sera has been proposed. Gold electrode was electrochemically modified with IrOx thin film and simultaneously functionalized with protein A (PA) to bind anti-CEA antibodies in an orientated way. It has been found that the antibody loading amount was dependent on the PA concentration and the deposition time of IrOx matrix. Under the optimized experimental conditions, the electron transfer resistances obtained were linearly related to the CEA concentration ranging from 36.2 to 460.0 ng/mL, with a detection limit of 28.0 ng/mL. Analytical results of clinical samples from cancer patients show that the proposed immunoassay is reasonably comparable with the chemiluminescence immunoassay (CLIA), indicating the feasibility of using the proposed method for CEA immunoassay in clinical laboratory.

  4. Separation of natural antioxidants using PDMS electrophoresis microchips coupled with amperometric detection and reverse polarity.

    Science.gov (United States)

    Lucca, Bruno Gabriel; Lunte, Susan Marie; Tomazelli Coltro, Wendell Karlos; Ferreira, Valdir Souza

    2014-12-01

    This report describes the use of PDMS ME coupled with amperometric detection for rapid separation of ascorbic, gallic , ferulic, p-coumaric acids using reverse polarity. ME devices were fabricated in PDMS by soft lithography and detection was accomplished using an integrated carbon fiber working electrode aligned in the end-channel configuration. Separation and detection parameters were investigated and the best conditions were obtained using a run buffer consisting of 5 mM phosphate buffer (pH 6.9) and a detection voltage of 1.0 V versus Ag/AgCl reference electrode. All compounds were separated within 70 s using gated injection mode with baseline resolution and separation efficiencies between 1200 and 9000 plates. Calibration curves exhibited good linearity and the LODs achieved ranged from 1.7 to 9.7 μM. The precision for migration time and peak height provided maximum values of 4% for the intrachip studies. Lastly, the analytical method was successfully applied for the analysis of ascorbic and gallic acids in commercial beverage samples. The results achieved using ME coupled with amperometric detection were in good agreement with the values provided by the supplier. Based on the data reported here, the proposed method shows suitability to be applied for the routine analysis of beverage samples.

  5. Hot-wire amperometric monitoring of flowing streams.

    Science.gov (United States)

    Wang, J; Jasinski, M; Flechsig, G U; Grundler, P; Tian, B

    2000-01-10

    This paper describes the design of a hot-wire electrochemical flow detector, and the advantages accrued from the effects of locally increased temperature, mainly thermally induced convection, upon the amperometric monitoring of flowing streams. A new hydrodynamic modulation voltammetric approach is presented, in which the solution flow rate remains constant while the temperature of the working electrode is modulated. Factors influencing the response, including the flow rate, temperature pulse, or applied potential, have been investigated. The hot-wire operation results also in a significant enhancement of the flow injection amperometric response. The minimal flow rate dependence observed with the heated electrode should benefit the on-line monitoring of streams with fluctuated natural convection, as well as various in-situ remote sensing applications.

  6. Electrochemical immunosensor with NiAl-layered double hydroxide/graphene nanocomposites and hollow gold nanospheres double-assisted signal amplification.

    Science.gov (United States)

    Qiao, Lu; Guo, Yemin; Sun, Xia; Jiao, Yancui; Wang, Xiangyou

    2015-08-01

    A sensitive electrochemical immunosensor based on NiAl-layered double hydroxide/graphene nanocomposites (NiAl-LDH/G) and hollow gold nanospheres (HGNs) was proposed for chlorpyrifos detection. The NiAl-LDH/G was prepared using a conventional coprecipitation process and reduction of the supporting graphene oxide. Subsequently, the nanocomposites were dispersed with chitosan (CS). The NiAl-LDH/G possessed good electrochemical behavior and high binding affinity to the electrode. The high surface areas of HGNs and the vast aminos and hydroxyls of CS provided a platform for the covalently crosslinking of antibody. Under optimal conditions, the immunosensor exhibited a wide linear range from 5 to 150 μg/mL and from 150 to 2 μg/mL, with a detection limit of 0.052 ng/mL. The detection results showed good agreement with standard gas chromatography method. The constructed immunosensor exhibited good reproducibility, high specificity, acceptable stability and regeneration performance, which provided a new promising tool for chlorpyrifos detection in real samples.

  7. Implementation of a SPR immunosensor for the simultaneous detection of the 22K and 20K hGH isoforms in human serum samples.

    Science.gov (United States)

    de Juan-Franco, Elena; Rodríguez-Frade, J M; Mellado, M; Lechuga, Laura M

    2013-09-30

    We have implemented a Surface Plasmon Resonance (SPR) immunosensor based on a sandwich assay for the simultaneous detection of the two main hGH isoforms, of 22 kDa (22K) and 20 kDa (20K). An oriented-antibody sensor surface specific for both hormone isoforms was assembled by using the biotin-streptavidin system. The immunosensor functionality was checked for the direct detection of the 22K hGH isoform in buffer, which gave high specificity and reproducibility (intra and inter-assay mean coefficients of variation of 8.23% and 9% respectively). The selective determination of the 22K and 20K hGH isoforms in human serum samples in a single assay was possible by using two specific anti-hGH monoclonal antibodies. The detection limit for both hormone isoforms was 0.9 ng mL(-1) and the mean coefficient of variation was below 7.2%. The excellent reproducibility and sensitivity obtained indicate the high performance of this immunosensor for implementing an anti-doping test.

  8. A novel electrochemical immunosensor using β-cyclodextrins functionalized silver supported adamantine-modified glucose oxidase as labels for ultrasensitive detection of alpha-fetoprotein.

    Science.gov (United States)

    Gao, Jian; Ma, Hongmin; Lv, Xiaohui; Yan, Tao; Li, Na; Cao, Wei; Wei, Qin

    2015-09-17

    In this work, a novel sandwich-type electrochemical immunosensor based on host-guest interaction was fabricated for the detection of alpha-fetoprotein (AFP). Due to the large specific surface area of multiwalled carbon nanotubes and the unique supramolecular recognition ability of β-cyclodextrins, ferrocenecarboxylic acid (Fc) was incorporated into this sensor platform by host-guest interaction to generate an electrochemical signal. And β-cyclodextrins functionalized silver supported adamantine-modified glucose oxidase (GOD-CD-Ag), was used as a label to improve the analytical performance of the immunosensor by the dual amplification strategy. The obtained GOD-CD-Ag conjugates could convert glucose into gluconic acid with the formation of hydrogen peroxide (H2O2). And then silver nanoparticles could in situ catalyze the reduction of the generated H2O2, dramatically improving the oxidation reaction of Fc. The developed immunosensor shows a wide linear calibration range from 0.001 to 5.0 ng/mL with a low detection limit (0.2 pg/mL) for the detection of AFP. The method, with ideal reproducibility and selectivity, has a wide application prospect in clinical research.

  9. Nanosilver-penetrated polyion graphene complex membrane for mediator-free amperometric immunoassay of alpha-fetoprotein using nanosilver-coated silica nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Tang Juan [Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, Department of Chemistry, Fuzhou University, Fuzhou 350108 (China); Tang Dianping, E-mail: dianping.tang@fzu.edu.c [Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, Department of Chemistry, Fuzhou University, Fuzhou 350108 (China); Su Biling; Li Qunfang; Qiu Bin [Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, Department of Chemistry, Fuzhou University, Fuzhou 350108 (China); Chen Guonan, E-mail: gnchen@fzu.edu.c [Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, Department of Chemistry, Fuzhou University, Fuzhou 350108 (China)

    2011-04-15

    Research highlights: {yields} We fabricate a polyion graphene complex membrane-based immunosensing platform for sensitive electrochemical immunoassay of alpha-fetoprotein. {yields} Nanosilver-coated silica nanocomposites as bionanolabels. {yields} Graphene nanosheets, single-stranded DNA and silver nanoparticles as matrices. {yields} Direct electron transfer without electron mediator. {yields} Analysis of real samples and method comparison. - Abstract: A facile and sensitive mediator-free electrochemical immunosensor for detection of alpha-fetoprotein (AFP) was designed by using nanosilver-coated silica nanoparticles (Ag-SiO{sub 2}) as bionanolabels. To construct such an electrochemical immunosensor, silver ions/single-stranded DNA/graphene nanosheets were initially immobilized on a gold electrode in turn, then silver ions were in situ reduced to silver nanoparticles with the aid of NaBH{sub 4}, and anti-AFP antibodies conjugated to silver nanoparticles were used. In the presence of AFP analyte, the sandwiched immunocomplex was formed on the electrode surface by using horseradish peroxidase-anti-AFP conjugate-labeled Ag-SiO{sub 2} (HRP-anti-AFP-Ag-SiO{sub 2}) as secondary antibodies. Compared with pure silver nanoparticles, Ag-SiO{sub 2} nanocomposites could provide a large room for the immobilization of HRP-anti-AFP, and improve the electrochemical responses of the immunosensor. Meanwhile, the presence of highly conductive graphene nanosheets and silver nanoparticles provided a good pathway for electron transfer. Under optimal conditions, the immunosensor exhibited good electrochemical responses toward AFP ranging from 0.3 to 200 ng/mL with a detection limit (LOD) of 0.05 ng/mL (at 3{sigma}) in pH 6.0 PBS-H{sub 2}O{sub 2} system. Intra- and inter-assay displayed good precisions with coefficient of variation below 9.5%. In addition, the method was evaluated with 23 clinical serum samples, receiving good correlation with results from commercially available

  10. Antibody immobilization on a nanoporous aluminum surface for immunosensor development

    Science.gov (United States)

    Chai, Changhoon; Lee, Jooyoung; Park, Jiyong; Takhistov, Paul

    2012-12-01

    A method of antibody (Ab) immobilization on a nanoporous aluminum surface for an electrochemical immunosensor is presented. To achieve good attachment and stability of Ab on an aluminum surface, aluminum was silanized with 3-aminopropyltryethoxysilane (APTES), and then covalently cross-linked to self-assembled layers (SALs) of APTES. Both the APTES concentration and the silanization time affected the formation of APTES-SALs as Ab immobilization. The formation of APTES-SALs was confirmed using the water contact angle on the APTES-SALs surface. The reactivity of APTES-SALs with Ab was investigated by measuring the fluorescence intensity of fluorescein isothiocyanate-labeled Ab-immobilized on the aluminum surface. Silanization of aluminum in 2% APTES for 4 h resulted in higher water contact angles and greater amounts of immobilized Ab than other APTES concentrations or silanization times. More Ab was immobilized on the nanoporous surface than on a planar aluminum surface. Electrochemical immunosensors developed on the nanoporous aluminum via the Ab immobilization method established in this study responded functionally to the antigen concentration in the diagnostic solution.

  11. A highly sensitive impedimetric label free immunosensor for Ochratoxin measurement in cocoa beans.

    Science.gov (United States)

    Malvano, Francesca; Albanese, Donatella; Pilloton, Roberto; Di Matteo, Marisa

    2016-12-01

    In this work the development and optimization of an impedimetric label free immunosensor for the detection of Ochratoxin A (OTA) is reported. Two antibody immobilization methods (oriented and not oriented) were compared highlighting a lower limit of detection (5pg/ml) for the not oriented immobilization but a closer linear range in contrast to oriented anti-OTA immunosensors which showed linearity in the range of 0.01-5ng/mL OTA. The analysis of the Atomic Force Microscopy (AFM) images showed two different nanostructures indicating that the use of oriented immobilization created a more ordered and highly dense antibody surface. Finally the oriented immunosensor was used to quantify OTA in spiked cocoa bean samples and the results were compared with those registered with competitive ELISA kit. The immunosensor was sensitive to OTA lower than 2μg/kg that represents the lower acceptable limit of OTA established by European legislation for the common food products.

  12. Studies towards the development of label-free AC impedimetric immunosensors for healthcare and food quality control

    OpenAIRE

    Tsekenis, Georgios

    2008-01-01

    This thesis describes work focused towards the fabrication and characterisation of immunosensor platforms for the label-free detection of analytes of importance in the health and food industries. Due to their low unit cost and ease of fabrication, the immunosensor market has significantly increased recently, resulting in a constant demand for new immunosensor applications. Within this thesis, therefore, a novel fabrication protocol is reported towards the production of immunose...

  13. CHARACTERISTICS AND OPTIMAL WORKING CONDITIONS OF AMPEROMETRIC BIOSENSOR FOR ADENOSINE TRIPHOSPHATE DETERMINATION

    Directory of Open Access Journals (Sweden)

    Kucherenko I. S.

    2014-02-01

    Full Text Available Analytical characteristics of a biosensor based on glucose oxidase and hexokinase and intended for ATP determination were studied. Platinum disc electrodes were used as amperometric transducers. Range of working potentials for biosensor functioning was shown. An optimal time of enzymes immobilization was determined. Optimal conditions for biosensor functioning during work with biological fluids were selected. Biosensor work in three buffer solutions (PBS, tris and HEPES was investigated and it was shown that it was possible to obtain various operational characteristics of the biosensor depending on tasks that are assigned to it by varying the composition of sample. Reproducibility of biosensor responses to ATP and glucose during a day and of biosensor preparation was shown. The proposed biosensor can be further used for analysis of glucose and ATP content in water solutions.

  14. A screen-printed, amperometric biosensor for the determination of organophosphorus pesticides in water samples

    Institute of Scientific and Technical Information of China (English)

    Junfeng Dou; Fuqiang Fan; Aizhong Ding; Lirong Cheng; Raju Sekar; Hongting Wang; Shuairan Li

    2012-01-01

    An amperometric biosensor based on screen-printed electrodes (SPEs) was developed for the determination of organophosphorus pesticides in water samples.The extent of acetylcholinesterase (AChE) deactivation was determined and quantified for pesticideconcentrations in water samples.An enzyme immobilization adsorption procedure and polyacrylamide gel matrix polymerization were used for fabrication of the biosensor,with minimal losses in enzyme activity.The optimal conditions for enzyme catalytic reaction on the SPEs surfaces were acetylthiocholine chloride (ATChC(1)) concentration of 5 mmol/L,pH 7 and reaction time of 4 min.The detectionlimits for three organophosphorus pesticides (dichlorvos,monocrotophs and parathion) were in the range of 4 to 7 μg/L when an AChE amount of 0.1 U was used for immobilization.

  15. APPLICATIONS OF ELECTROCHEMICAL IMMUNOSENSORS TO ENVIRONMENTAL MONITORING

    Science.gov (United States)

    This paper discusses basic electrochemical immunoassay technology. Factors limiting the practical application of antibodies to anlaytical problems are also presented. It addresses the potential use of immunoassay methods based on electrochemical detection for the analysis of env...

  16. Cellulose antibody films for highly specific evanescent wave immunosensors

    Science.gov (United States)

    Hartmann, Andreas; Bock, Daniel; Jaworek, Thomas; Kaul, Sepp; Schulze, Matthais; Tebbe, H.; Wegner, Gerhard; Seeger, Stefan

    1996-01-01

    For the production of recognition elements for evanescent wave immunosensors optical waveguides have to be coated with ultrathin stable antibody films. In the present work non amphiphilic alkylated cellulose and copolyglutamate films are tested as monolayer matrices for the antibody immobilization using the Langmuir-Blodgett technique. These films are transferred onto optical waveguides and serve as excellent matrices for the immobilization of antibodies in high density and specificity. In addition to the multi-step immobilization of immunoglobulin G(IgG) on photochemically crosslinked and oxidized polymer films, the direct one-step transfer of mixed antibody-polymer films is performed. Both planar waveguides and optical fibers are suitable substrates for the immobilization. The activity and specificity of immobilized antibodies is controlled by the enzyme-linked immunosorbent assay (ELISA) technique. As a result reduced non-specific interactions between antigens and the substrate surface are observed if cinnamoylbutyether-cellulose is used as the film matrix for the antibody immobilization. Using the evanescent wave senor (EWS) technology immunosensor assays are performed in order to determine both the non-specific adsorption of different coated polymethylmethacrylat (PMMA) fibers and the long-term stability of the antibody films. Specificities of one-step transferred IgG-cellulose films are drastically enhanced compared to IgG-copolyglutamate films. Cellulose IgG films are used in enzymatic sandwich assays using mucine as a clinical relevant antigen that is recognized by the antibodies BM2 and BM7. A mucine calibration measurement is recorded. So far the observed detection limit for mucine is about 8 ng/ml.

  17. A wafer-level liquid cavity integrated amperometric gas sensor with ppb-level nitric oxide gas sensitivity

    Science.gov (United States)

    Gatty, Hithesh K.; Stemme, Göran; Roxhed, Niclas

    2015-10-01

    A miniaturized amperometric nitric oxide (NO) gas sensor based on wafer-level fabrication of electrodes and a liquid electrolyte chamber is reported in this paper. The sensor is able to detect NO gas concentrations of the order of parts per billion (ppb) levels and has a measured sensitivity of 0.04 nA ppb-1 with a response time of approximately 12 s. A sufficiently high selectivity of the sensor to interfering gases such as carbon monoxide (CO) and to ammonia (NH3) makes it potentially relevant for monitoring of asthma. In addition, the sensor was characterized for electrolyte evaporation which indicated a sensor operation lifetime allowing approximately 200 measurements.

  18. Ring-Resonator/Sol-Gel Interferometric Immunosensor

    Science.gov (United States)

    Bearman, Gregory; Cohen, David

    2007-01-01

    A proposed biosensing system would be based on a combination of (1) a sensing volume containing antibodies immobilized in a sol-gel matrix and (2) an optical interferometer having a ring resonator configuration. The antibodies would be specific to an antigen species that one seeks to detect. In the ring resonator of the proposed system, light would make multiple passes through the sensing volume, affording greater interaction length and, hence, greater antibody- detection sensitivity.

  19. Cleanup and analysis of sugar phosphates in biological extracts by using solid phase extraction and anion-exchange chromatography with pulsed amperometric detection

    DEFF Research Database (Denmark)

    Smith, Hans Peter; Cohen, A.; Buttler, T.

    1998-01-01

    A cleanup method based on anion-exchange solid-phase extraction (SPE) was developed to render biological extracts suitable for the analysis of hexose phosphates with a modified anion-exchange chromatography method and pulsed amperometric detection. The method was applied to cell extracts of Sacch......A cleanup method based on anion-exchange solid-phase extraction (SPE) was developed to render biological extracts suitable for the analysis of hexose phosphates with a modified anion-exchange chromatography method and pulsed amperometric detection. The method was applied to cell extracts...... of Saccharomyces cerevisiae obtained by using cold methanol as quenching agent and chloroform as extraction solvent. It was shown that pretreatment of the cell extract with SPE markedly improved the quality of the liquid chromatography analysis with recoveries of the sugar phosphates close to 100%. Furthermore...

  20. Efficient streptavidin-functionalized nitrogen-doped graphene for the development of highly sensitive electrochemical immunosensor.

    Science.gov (United States)

    Yang, Zhanjun; Lan, Qingchun; Li, Juan; Wu, Jiajia; Tang, Yan; Hu, Xiaoya

    2017-03-15

    In this work, an efficient and universal streptavidin-functionalized nitrogen-doped graphene (NG) was for the first time proposed and used to develop a highly sensitive electrochemical immunosensor for the detection of tumor markers. Transmission electron microscopy, electrochemical impedance spectrum, static water contact measurement, and cyclic voltammetry were used to characterize the streptavidin-functionalized NG platform and immunosensor. The biofunctionalized NG showed excellent hydrophilicity, larger specific surface area, and high electrochemical activity. These properties of the platform enhanced the loading capacity of proteins, and retained the bioactivity of the immobilized proteins, and thus remarkably improved the sensitivity of the immunosensor. Using carcinoembryonic antigen (CEA) as model analyte, the proposed immunosensor demonstrated a wide linear range of 0.02-12ngmL(-1) with a low detection limit of 0.01ngmL(-1). The CEA immunosensor could be applied to detect human serum samples with satisfactory results. The streptavidin-functionalized NG material provided an universal and promising platform for the electrochemical immunosensing applications.

  1. Impedimetric Label-Free Immunosensor on Disposable Modified Screen-Printed Electrodes for Ochratoxin A.

    Science.gov (United States)

    Malvano, Francesca; Albanese, Donatella; Crescitelli, Alessio; Pilloton, Roberto; Esposito, Emanuela

    2016-06-30

    An impedimetric label-free immunosensor on disposable screen-printed carbon electrodes (SPCE) for quantitative determination of Ochratoxin A (OTA) has been developed. After modification of the SPCE surface with gold nanoparticles (AuNPs), the anti-OTA was immobilized on the working electrode through a cysteamine layer. After each coating step, the modified surfaces were characterized by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The capacitance was chosen as the best parameter that describes the reproducible change in electrical properties of the electrode surface at different OTA concentrations and it was used to investigate the analytical parameters of the developed immunosensor. Under optimized conditions, the immunosensor showed a linear relationship between 0.3 and 20 ng/mL with a low detection limit of 0.25 ng/mL, making it suitable to control OTA content in many common food products. Lastly, the immunosensor was used to measure OTA in red wine samples and the results were compared with those registered with a competitive ELISA kit. The immunosensor was sensitive to OTA lower than 2 μg/kg, which represents the lower acceptable limit of OTA established by European legislation for common food products.

  2. Impedimetric Label-Free Immunosensor on Disposable Modified Screen-Printed Electrodes for Ochratoxin A

    Directory of Open Access Journals (Sweden)

    Francesca Malvano

    2016-06-01

    Full Text Available An impedimetric label-free immunosensor on disposable screen-printed carbon electrodes (SPCE for quantitative determination of Ochratoxin A (OTA has been developed. After modification of the SPCE surface with gold nanoparticles (AuNPs, the anti-OTA was immobilized on the working electrode through a cysteamine layer. After each coating step, the modified surfaces were characterized by cyclic voltammetry (CV and electrochemical impedance spectroscopy (EIS. The capacitance was chosen as the best parameter that describes the reproducible change in electrical properties of the electrode surface at different OTA concentrations and it was used to investigate the analytical parameters of the developed immunosensor. Under optimized conditions, the immunosensor showed a linear relationship between 0.3 and 20 ng/mL with a low detection limit of 0.25 ng/mL, making it suitable to control OTA content in many common food products. Lastly, the immunosensor was used to measure OTA in red wine samples and the results were compared with those registered with a competitive ELISA kit. The immunosensor was sensitive to OTA lower than 2 μg/kg, which represents the lower acceptable limit of OTA established by European legislation for common food products.

  3. A green synthetic strategy of nickel hexacyanoferrate nanoparticals supported on the graphene substrate and its non-enzymatic amperometric sensing application

    Science.gov (United States)

    xue, Zhonghua; He, Nan; Rao, Honghong; Hu, Chenxian; Wang, Xiaofen; Wang, Hui; Liu, Xiuhui; Lu, Xiaoquan

    2017-02-01

    Rapid glucose detection is a key requirement for both diagnosis and treatment of diabetes. A facile and green strategy to achieve spherical-shaped nickel hexacyanoferrate (NiHCF) nanoparticals supported on electrochemical reduction graphene oxide by using electrochemical cyclic voltammetry is explored. As a sensing substrate, electrochemical reduction graphene oxide deposited on a glassy carbon electrode surface exhibited obvious positive effect on the electrodeposition of NiHCF nanoparticals with spherical structure and thus effectively improved the electrical conductivity and electrochemical sensing of the proposed amperometric sensor. Proof-concept experiments demonstrated that the proposed nanocomposites modified electrode exhibited excellent sensitivity toward glucose oxidation as well as with a satisfying detection limit of 0.11 μM. More importantly, we also explore that as a simple, green and facile method, electrochemical technology can be employed and provide a new strategy for developing GO and metal hexacyanoferrate based amperometric sensing platform toward glucose and other biomolecules.

  4. Visual and efficient immunosensor technique for advancing biomedical applications of quantum dots on Salmonella detection and isolation

    Science.gov (United States)

    Tang, Feng; Pang, Dai-Wen; Chen, Zhi; Shao, Jian-Bo; Xiong, Ling-Hong; Xiang, Yan-Ping; Xiong, Yan; Wu, Kai; Ai, Hong-Wu; Zhang, Hui; Zheng, Xiao-Li; Lv, Jing-Rui; Liu, Wei-Yong; Hu, Hong-Bing; Mei, Hong; Zhang, Zhen; Sun, Hong; Xiang, Yun; Sun, Zi-Yong

    2016-02-01

    It is a great challenge in nanotechnology for fluorescent nanobioprobes to be applied to visually detect and directly isolate pathogens in situ. A novel and visual immunosensor technique for efficient detection and isolation of Salmonella was established here by applying fluorescent nanobioprobes on a specially-designed cellulose-based swab (a solid-phase enrichment system). The selective and chromogenic medium used on this swab can achieve the ultrasensitive amplification of target bacteria and form chromogenic colonies in situ based on a simple biochemical reaction. More importantly, because this swab can serve as an attachment site for the targeted pathogens to immobilize and immunologically capture nanobioprobes, our mAb-conjugated QD bioprobes were successfully applied on the solid-phase enrichment system to capture the fluorescence of targeted colonies under a designed excitation light instrument based on blue light-emitting diodes combined with stereomicroscopy or laser scanning confocal microscopy. Compared with the traditional methods using 4-7 days to isolate Salmonella from the bacterial mixture, this method took only 2 days to do this, and the process of initial screening and preliminary diagnosis can be completed in only one and a half days. Furthermore, the limit of detection can reach as low as 101 cells per mL Salmonella on the background of 105 cells per mL non-Salmonella (Escherichia coli, Proteus mirabilis or Citrobacter freundii, respectively) in experimental samples, and even in human anal ones. The visual and efficient immunosensor technique may be proved to be a favorable alternative for screening and isolating Salmonella in a large number of samples related to public health surveillance.It is a great challenge in nanotechnology for fluorescent nanobioprobes to be applied to visually detect and directly isolate pathogens in situ. A novel and visual immunosensor technique for efficient detection and isolation of Salmonella was established here

  5. Poly 3,4-ethylenedioxythiophene as an entrapment support for amperometric enzyme sensor

    OpenAIRE

    Fabiano, Silvia; Tran-Minh, Canh; Piro, Benoît; Anh Dang, Lan; Chau Pham, Minh; Vittori, Olivier

    2002-01-01

    International audience; A conducting polymer of poly 3,4-ethylenedioxythiophene (PEDT) was used as a matrix for entrapment of enzymes onto a platinum electrode surface in order to construct amperometric biosensors. Glucose oxidase (GOD) was used as an example, and it was entrapped in the polymer during the electrochemical polymerization. Glucose in oxygenated solutions was tested by amperometric measurements at +650 mV (vs. SCE) in a batch system. The influence of several experimental paramet...

  6. Direct immobilization of antibodies on a new polymer film for fabricating an electrochemical impedance immunosensor.

    Science.gov (United States)

    Zhang, Xiangyang; Shen, Guangyu; Shen, Youming; Yin, Dan; Zhang, Chunxiang

    2015-09-15

    A new polymer bearing aldehyde groups was designed and synthesized by grafting 4-pyridinecarboxaldehyde onto poly(epichlorohydrin). Antibodies can be directly immobilized on the surface of the polymer film through the covalent bonding of aldehyde groups of the film with amino groups of antibodies. In this study, human immunoglobulin G (IgG) was used as a model analyte for the fabrication of an electrochemical impedance immunosensor. Using the proposed immunosensor, IgG in the range from 0.1 to 80 ng ml(-1) was detected with a detection limit of 0.07 ng ml(-1) (signal/noise [S/N]=3). In addition, the electrochemical impedance immunosensor displays good stability and reproducibility.

  7. Development of SPR Immunosensor by Indirect Competitive Method for Rapid and Highly Sensitive Salivary Cortisol Detection

    Directory of Open Access Journals (Sweden)

    Yusuke eTahara

    2014-05-01

    Full Text Available The monitoring of salivary cortisol as a key biomarker of an individual’s stress response has been increasingly focused on. This paper describes the development of a novel cortisol biosensor based on an indirect competitive method using a surface plasmon resonance (SPR immunosensor. The surface of a Au chip was modified with PEG6-COOH aromatic dialkanethiol self-assembled monolayers (SAMs and hydrocortisone 3-(O-carboxymethyl oxime (hydrocortisone 3-CMO as a cortisol analogue. A detection limit of 38 ppt range with a measurement range of 10 ppt to 100 ppb was accomplished without the incubation of a mixing solution consisting of standard cortisol and an anti-cortisol antibody, and the response time was 5 min from the sample injection. We experimentally compared our biosensor with a commercialized salivary cortisol enzyme-linked immunosorbent assay (ELISA kit using human saliva samples. It was found that the results obtained by the cortisol biosensor had a high correlation with those obtained by ELISA assay (R = 0.96. Our findings indicate the potential utility of the cortisol biosensor for on-site diagnosis and bedside point-of-care testing (POCT from bedside testing.

  8. Surface plasmon resonance immunosensor for the detection of Salmonella typhi antibodies in buffer and patient serum.

    Science.gov (United States)

    Gupta, Garima; Sharma, P K; Sikarwar, B; Merwyn, S; Kaushik, S; Boopathi, M; Agarwal, G S; Singh, Beer

    2012-01-01

    Surface plasmon resonance (SPR) immunosensor using 4-mercaptobenzoic acid (4-MBA) modified gold SPR chip was developed first time for the detection of flagellin specific antibodies of Salmonella typhi (S. typhi). Flagellin protein of S. typhi was prepared by recombinant DNA technology. The modification of gold chip with 4-MBA was in-situ characterized by SPR and electrochemical impedance spectroscopy. By using kinetic evaluation software, K(D) and B(max) values were calculated and found to be 26.3 fM and 62.04 m°, respectively, for the immobilized monoclonal antibody (Moab) of recombinant flagellin (r-fla) protein of S. typhi (r-fla S. typhi). In addition, thermodynamic parameters such as ΔG, ΔH and ΔS were determined first time for r-fla S. typhi and Moab of r-fla S. typhi interactions and the values revealed the interaction between r-fla S. typhi and Moab of r-fla S. typhi as spontaneous, endothermic and entropy driven one. Moreover, healthy human serum samples and patient sera (Widal positive and Widal negative) were subjected to SPR analysis. The present SPR based approach provides an alternative way for S. typhi detection in less than 10 min.

  9. Sensitivity Improvement of an Impedimetric Immunosensor Using Functionalized Iron Oxide Nanoparticles

    Directory of Open Access Journals (Sweden)

    Imen Hafaid

    2009-01-01

    Full Text Available This work has explored the development of impedimetric immunosensors based on magnetic iron nanoparticles (IrNP functionalized with streptavidin to which a biotinylated FAB part of the antibody has been bound using a biotin-streptavidin interaction. SPR analysis shows a deviation on the measured (angle during antigen-antibody recognition whereas label free detection using by EIS allows us to monitor variation of polarization resistance. Before detection, layers were analyzed by FTIR and AFM. Compared to immobilization of antibody on bare gold surface using aminodecanethiol SAM, antibody immobilization on nanoparticles permitted to reach lower detection limit: 500 pg/ml instead of 1 ng/ml to in the case of EIS and 300 ng/ml instead of 4.5 μg/ml in the case of SPR. Thus, it permitted to improve the sensitivity: from 257.3  Ω⋅cm2⋅μg−1⋅ml to 1871 Ω⋅cm2⋅μg−1⋅ml in the case of EIS and from 0.003°μg−1⋅ml to 0.094°μg−1⋅ml in the case of SPR.

  10. An electrochemical immunosensor for carcinoembryonic antigen enhanced by self-assembled nanogold coatings on magnetic particles.

    Science.gov (United States)

    Li, Jianping; Gao, Huiling; Chen, Zhiqiang; Wei, Xiaoping; Yang, Catherine F

    2010-04-14

    A quick and reproducible electrochemical-based immunosensor technique, using magnetic core/shell particles that are coated with self-assembled multilayer of nanogold, has been developed. Magnetic particles that are structured from Au/Fe(3)O(4) core-shells were prepared and aminated after a reaction between gold and thiourea, and additional multilayered coatings of gold nanoparticles were assembled on the surface of the core/shell particles. The carcinoembryonic antibody (anti-CEA) was immobilized on the modified magnetic particles, which were then attached on the surface of solid paraffin carbon paste electrode (SPCE) by an external magnetic field. This is an assembly of a novel immuno biosensor for carcinoembryonic antigen (CEA). The sensitivity and response features of this immunoassay are significantly affected by the surface area and the biological compatibility of the multilayered nanogold. The linear range for the detection of CEA was from 0.005 to 50 ng mL(-1) and the limit of detection (LOD) was 0.001 ng mL(-1). The LOD is approximately 500 times more sensitive than that of the traditional enzyme-linked immunosorbent assay for CEA detection.

  11. Magneto immunosensor for gliadin detection in gluten-free foodstuff: towards food safety for celiac patients.

    Science.gov (United States)

    Laube, T; Kergaravat, S V; Fabiano, S N; Hernández, S R; Alegret, S; Pividori, M I

    2011-09-15

    Gliadin is a constituent of the cereal protein gluten, responsible for the intolerance generated in celiac disease. Its detection is of high interest for food safety of celiac patients, since the only treatment known until now is a lifelong avoidance of this protein in the diet. Therefore, it is essential to have an easy and reliable method of analysis to control the contents in gluten-free foods. An electrochemical magneto immunosensor for the quantification of gliadin or small gliadin fragments in natural or pretreated food samples is described for the first time and compared to a novel magneto-ELISA system based on optical detection. The immunological reaction was performed on magnetic beads as solid support by the oriented covalent immobilization, of the protein gliadin on tosyl-activated beads. Direct, as well as indirect competitive immunoassays were optimized, achieving the best analytical performance with the direct competitive format. Excellent detection limits (in the order of μg L(-1)) were achieved, according to the legislation for gluten-free products. The matrix effect, as well as the performance of the assays was successfully evaluated using spiked gluten-free foodstuffs (skimmed milk and beer), obtaining excellent recovery values in the results.

  12. Detection of estradiol at an electrochemical immunosensor with a Cu UPD|DTBP-Protein G scaffold.

    Science.gov (United States)

    Liu, Xiaoqiang; Wang, Xinhai; Zhang, Jiamei; Feng, Heqing; Liu, Xiuhua; Wong, Danny K Y

    2012-05-15

    A copper monolayer was formed on a gold electrode surface via underpotential deposition (UPD) method to construct a Cu UPD|DTBP-Protein G immunosensor for the sensitive detection of 17β-estradiol. Copper UPD monolayer can minimize the non-specific adsorption of biological molecules on the immunosensor surface and enhance the binding efficiency between immunosensor surface and thiolated Protein G. The crosslinker DTBP (Dimethyl 3,3'-dithiobispropionimidate · 2HCl) has strong ability to immobilize Protein G molecules on the electrode surface and the immobilized Protein G provides an orientation-controlled binding of antibodies. A monolayer of propanethiol was firstly self-assembled on the gold electrode surface, and a copper monolayer was deposited via UPD on the propanethiol modified electrode. Propanethiol monolayer helps to stabilize the copper monolayer by pushing the formation and stripping potentials of the copper UPD monolayer outside the potential range in which copper monolayer can be damaged easily by oxygen in air. A droplet DTBP-Protein G was then applied on the modified electrode surface followed by the immobilization of estradiol antibody. Finally, a competitive immunoassay was conducted between estradiol-BSA (bovine serum albumin) conjugate and free estradiol for the limited binding sites of estradiol antibody. Square wave voltammetry (SWV) was employed to monitor the electrochemical reduction current of ferrocenemethanol and the SWV current decreased with the increase of estradiol-BSA conjugate concentration at the immunosensor surface. Calibration of immunosensors in waste water samples spiked with 17β-estradiol yielded a linear response up to ≈ 2200 pg mL(-1), a sensitivity of 3.20 μA/pg mL(-1) and a detection limit of 12 pg mL(-1). The favorable characteristics of the immunosensors such as high selectivity, sensitivity and low detection limit can be attributed to the Cu UPD|DTBP-Protein G scaffold.

  13. A Gold Nanoparticles Enhanced Surface Plasmon Resonance Immunosensor for Highly Sensitive Detection of Ischemia-Modified Albumin

    Directory of Open Access Journals (Sweden)

    Guang Li

    2013-09-01

    Full Text Available In this study a novel sensitive nanogold particle sensor enhancement based on mixed self-assembled monolayers was explored and used to construct a Surface Plasmon Resonance (SPR immunosensor to detect Ischemia Modified Albumin (IMA. Compared with a direct binding SPR assay at a limit of detection (LOD of 100 ng/L, gold nanoparticles (AuNPs of 10 nm dramatically improved the LOD of IMA to 10 ng/L. Meanwhile, no interfering substance that may lead to false positive results was identified. These results suggested that the SPR biosensor presented superior properties, and provided a simple label-free strategy to increase assay sensitivity for further acute coronary syndrome (ACS diagnosis.

  14. Sensitive immunoassay of human chorionic gonadotrophin based on multi-walled carbon nanotube-chitosan matrix.

    Science.gov (United States)

    Li, Na; Yuan, Ruo; Chai, Yaqin; Chen, Shihong; An, Haizhen

    2008-10-01

    A novel amperometric immunosensor for human chorionic gonadotropin (HCG) assay has been fabricated through incorporating toluidine blue (TB) and hemoglobin (Hb) on the multiwall carbon nanotube (MWNT)-chitosan (CS) modified glassy carbon electrode, followed by electrostatic adsorption of a conducting gold nanoparticles (nanogold) film as sensing interface. The MWNT-CS matrix provided a congenial microenvironment for the immobilization of biomolecules and promoted the electron transfer to enhance the sensitivity of the immunosensor. Due to the strong electrocatalytic properties of Hb and MWNT toward H(2)O(2), the Hb and MWNT significantly amplified the current signal of the antigen-antibody reaction. The immobilized toluidine blue as an electron transfer mediator exhibited excellent electrochemical redox property. After the immunosensor was incubated with HCG solution, the access of activity center of the Hb to toluidine blue was partly inhibited, which leaded to a linear decrease in the catalytic efficiency of the Hb to the oxidation of immobilized toluidine blue by H(2)O(2) over HCG concentration ranges from 0.8 to 500 mIU/mL. Under optimal condition, the detection limit for the HCG immunoassay was 0.3 mIU/mL estimated at a signal-to-noise ratio of 3. Moreover, the proposed immunosensor displayed a satisfactory stability and reproducibility.

  15. Fiber optic immunosensor for cross-linked fibrin concentration

    Science.gov (United States)

    Moskowitz, Samuel E.

    2000-08-01

    Working with calcium ions in the blood, platelets produce thromboplastin which transforms prothrombin into thrombin. Removing peptides, thrombin changes fibrinogen into fibrin. Cross-linked insoluble fibrin polymers are solubilized by enzyme plasmin found in blood plasma. Resulting D-dimers are elevated in patients with intravascular coagulation, deep venous thrombosis, pulmonary embolism, myocardial infarction, multiple trauma, cancer, impaired renal and liver functions, and sepsis. Consisting principally of a NIR 780 nm GaAlAs laser diode and a 800 nm avalanche photodiode (APD), the fiber-optic immunosensor can determined D-dimer concentration to levels detected by a second antibody which is labeled with NN 382 fluorescent dye. An evanescent wave traveling on an excitation optical fiber excites the antibody-antigen fluorophore complex. Concentration of cross-linked fibrin is directly proportional to the APD measured intensity of fluorescence. NIR fluorescence has advantages of low background interference, short fluorescence lifetime, and large difference between excitation and emission peaks. Competitive ELISA test for D-dimer concentration requires trained personnel performing a time consuming operation.

  16. Immunosensor Incorporating Anti-His (C-term IgG F(ab’ Fragments Attached to Gold Nanorods for Detection of His-Tagged Proteins in Culture Medium

    Directory of Open Access Journals (Sweden)

    Michal Wąsowicz

    2010-06-01

    Full Text Available Immunosensors based on gold electrodes (electrochemical or gold discs (optical modified with 1,6-hexanedithiol, gold nanorods and Anti-His (C-term monoclonal antibody F(ab’ fragment are described. The antigen detected by the sensing platform is a recombinant histidine-tagged silk proteinase inhibitor (rSPI2-His6. Electrochemical impedance spectroscopy (EIS and surface plasmon resonance (SPR techniques were used as methods for detection of the antigen. This approach allows to detect the antigen protein in concentration of 10 pg per mL (0.13 pM of culture medium. The immunosensor shows good reproducibility due to covalent immobilization of F(ab’ fragments to gold nanorods layer

  17. Novel impedimetric immunosensor for the detection and quantitation of Adenovirus using reduced antibody fragments immobilized onto a conducting copolymer surface.

    Science.gov (United States)

    Caygill, Rebecca L; Hodges, Christopher S; Holmes, Joanne L; Higson, Séamus P J; Blair, G Eric; Millner, Paul A

    2012-02-15

    The number of Adenovirus (Ad) infections detected in immunocompromised people has increased due to the number of patients receiving transplants, as well as the HIV pandemic. Ads cause life-threatening diseases specific to the infected organs of immunocompromised hosts, with discontinuation of immunosuppressive agents necessary to prevent morbidity. The methodology in this paper has been employed to develop a novel impedimetric based assay platform to detect and quantify human Ads, which is comparable in performance to current methods, such as ELISA and PCR, but is also less expensive and faster. Novel immunosensors have been fabricated using polyclonal antibodies raised against a human Ad (Ad5) capsid protein, which were selectively cleaved into antibody fragments by 2-mercaptoethylamine. The fragments were immobilized onto a functionalized conducting copolymer matrix comprising polyaniline and 2-aminobenzylamine. Fully fabricated sensors were incubated with two immunologically distinct serotypes of Ad, Ad5 and Ad3, with between 10 and 10(12)virus particles/mL prior to sensor interrogation. Electrochemical impedance spectroscopy was used to measure the charge transfer resistance of the sensors over a range of frequencies from 25 kHz to 0.1 Hz. Our data demonstrate that the immunosensors specifically detect, and differentiate between, closely related human Ad serotypes with a limit of detection of 10(3)virus particles/mL. In addition, atomic force microscopy was applied to study the sensor surface nanostructure. Future work looks to test virus containing clinical samples but this could be a viable and valuable alternative for point-of-care virus detection and quantification.

  18. An approach to conductometric immunosensor based on phthalocyanine thin film.

    Science.gov (United States)

    Sergeyeva, T A; Lavrik, N V; Rachkov, A E; Kazantseva, Z I; El'skaya, A V

    1998-03-01

    A new approach to conductometric biosensors utilizing iodine-sensitive phthalocyanine thin films has been proposed. The excellent sensitivity of the tetra-tert-butyl copper phthalocyanine (ttb-CuPc) to free iodine was used for the first time to detect a peroxidase-initiated reaction in an aqueous medium. To minimize the interfering effect of aqueous electrolytes on the impedance responses of the ttb-CuPc film itself, Au/Cr interdigitated planar electrodes bearing ttb-CuPc thin films were protected with hydrophobic gas-permeable membranes, namely thermally evaporated calixarene or plasma polymerized hexamethyldisiloxane films. Impedance spectroscopy data were analyzed in order to define the optimal operating frequency. An enzyme sensor with peroxidase immobilized in a cross-linked albumin matrix was tested. Its impedance responses were studied under variation of the substrate concentration, pH, ionic strength and buffer capacity. These results were used to define conditions for peroxidase-linked immunoassay in subsequent tests. With the developed sensor, concentrations of IgG in 0.2-2 micrograms/ml range were measured in a competitive mode with satisfactory accuracy. The detection of IgG in both test solutions and blood serum samples has been demonstrated.

  19. Feasibility of evanescent wave interferometer immunosensors for pesticide detection: chemical aspects

    NARCIS (Netherlands)

    Lechuga, L.M.; Lenferink, A.T.M.; Kooyman, R.P.H.; Greve, J.

    1995-01-01

    A waveguide Mach-Zehnder Interferometer (MZI) immunosensor has been developed which can detect, in a direct way, a minimum average layer growth thickness of the analyte of 2×10−3 nm (bound mass, I′=1×10−4 δ cm−2). The design and fabrication of the sensor and the experimental set-up are aimed at decr

  20. Rapid Determination of Phytophthora infestans sporangia Using a Surface Plasmon Resonance Immunosensor

    DEFF Research Database (Denmark)

    Skottrup, Peter; Nicolaisen, Mogens; Justesen, Annemarie Fejer

    2007-01-01

    Phytophthora infestans is the cause of late blight disease in potato and is an economically important pathogen worldwide. Early disease detection is important to implement disease control measures. In this study a surface plasmon resonance (SPR) immunosensor for detection of P. infestans sporangia...

  1. Rapid Determination of Phytophthora infestans sporangia Using a Surface Plasmon Resonance Immunosensor

    DEFF Research Database (Denmark)

    Skottrup, Peter Durand; Nicolaisen, Mogens; Justesen, Annemarie Fejer

    2007-01-01

    Phytophthora infestans is the cause of late blight disease in potato and is an economically important pathogen worldwide. Early disease detection is important to implement disease control measures. In this study a surface plasmon resonance (SPR) immunosensor for detection of P. infestans sporangi...

  2. Comparative study of random and oriented antibody immobilization techniques on the binding capacity of immunosensor.

    Science.gov (United States)

    Kausaite-Minkstimiene, A; Ramanaviciene, A; Kirlyte, J; Ramanavicius, A

    2010-08-01

    A comparative study of four different antibody immobilization techniques that are suitable for modification of surface plasmon resonance (SPR) chip (SPR-chip) is reported. Antibodies against human growth hormone (anti-HGH) were used as the model system. The evaluated SPR-chip modification techniques were (i) random immobilization of intact anti-HGH (intact-anti-HGH) via self-assembled monolayer (SAM) based on 11-mercaptoundecanoic acid (MUA); (ii) random immobilization of intact-anti-HGH within carboxymethyl dextran (CMD) hydrogel by direct covalent amine coupling technique; (iii) oriented coupling of intact-anti-HGH via Fc-fragment to protein-G layer assembled on SAM consisting of MUA (MUA/pG); (iv) oriented immobilization of fragmented anti-HGH antibodies (frag-anti-HGH) via their native thiol-groups directly coupled to the gold. To liberate these thiol groups, the intact-anti-HGH was chemically "divided" into two frag-anti-HGH fragments by chemical reduction with 2-mercaptoethylamine (2-MEA). Optimal concentration of 2-MEA for preparation of anti-HGH was 15 mM. The surface concentration of immobilized antibodies and the antigen binding capacity for all four differently modified SPR-chips was evaluated and compared. The maximum surface concentration of immobilized intact-anti-HGH was obtained by immobilizing the antibody within CMD-hydrogel. The maximal antigen binding capacity was obtained by SPR-chip based on intact-anti-HGH immobilized via MUA/pG. The immobilization based on application of frag-anti-HGH was found to be the most suitable for design of SPR-immunosensor for HGH detection, due to its sufficient antigen binding capacity, simplicity, and low cost in respect to the currently evaluated techniques.

  3. Novel signal amplification strategy for ultrasensitive sandwich-type electrochemical immunosensor employing Pd-Fe3O4-GS as the matrix and SiO2 as the label.

    Science.gov (United States)

    Wang, Yulan; Ma, Hongmin; Wang, Xiaodong; Pang, Xuehui; Wu, Dan; Du, Bin; Wei, Qin

    2015-12-15

    An ultrasensitive sandwich-type electrochemical immunosensor based on a novel signal amplification strategy was developed for the quantitative determination of human immunoglobulin G (IgG). Pd nanocubes functionalized magnetic graphene sheet (Pd-Fe3O4-GS) was employed as the matrix to immobilize the primary antibodies (Ab1). Owing to the synergetic effect between Pd nanocubes and magnetic graphene sheet (Fe3O4-GS), Pd-Fe3O4-GS can provide an obviously increasing electrochemical signal by electrochemical catalysis towards hydrogen peroxide (H2O2). Silicon dioxide (SiO2) was functionalized as the label to conjugate with the secondary antibodies (Ab2). Due to the larger steric hindrance of the obtained conjugate (SiO2@Ab2), the sensitive decrease of the electrochemical signal can be achieved after the specific recognition between antibodies and antigens. In this sense, this proposed immunosensor can achieve a high sensitivity, especially in the presence of low concentrations of IgG. Under optimum conditions, the proposed immunosensor offered an ultrasensitive and specific determination of IgG down to 3.2 fg/mL. This immunoassay method would open up a new promising platform to detect various tumor markers at ultralow levels for early diagnoses of different cancers.

  4. A study of biofunctionalized silica nanospring surface for immunosensor applications

    Science.gov (United States)

    Timalsina, Yukta P.; McIlroy, David N.

    2012-02-01

    A study of biofunctionalized VANS (vertically aligned (silica) nanospring) surface for immunosensor applications is presented. VANS surface treated with 3-aminopropyltriethoxysilane (APTES) leaves a primary amine groups on the VANS surface. Glutaraldehyde (GA) reacts with APTES modified VANS surface forming imine bonds at one end of glutaraldehyde, leaving aldehyde groups at the other end to react with the antibody. X-ray photoelectron study verifies each step of VANS surface functionalization. A goat anti mouse antibody (GαM IgG I) is immobilized as a biorecognition layer on the APTES-GA modified surface and targeted to mouse IgG. It is investigated that mouse IgG captured from the solution phase specifically binds to goat anti mouse IgG on APTES-GA- GαM IgG I. Then layer of GαM IgG II attached to the APTES-GA- GαM IgG I-mouse IgG surface reacts only when there is mouse IgG instead of rabbit IgG. A modeling of a resistor-inductor-capacitor (RLC) circuit of impedance spectra measured after the addition of successive layer indicates the these biological layers behave as insulating layers. It is explored that there is a greater magnitude of change between successive bio-layers below 10 kHz. Changes in the magnitudes of the elements of the RLC equivalent circuit indicate that the addition of biological layers impedes ionic motion thereby changing the effective dielectric response by the biomolecule polarization.

  5. Amperometric microsensor for direct probing of ascorbic acid in human gastric juice.

    Science.gov (United States)

    Hutton, Emily A; Pauliukaitė, Rasa; Hocevar, Samo B; Ogorevc, Božidar; Smyth, Malcolm R

    2010-09-30

    This article reports on a novel microsensor for amperometric measurement of ascorbic acid (AA) under acidic conditions (pH 2) based on a carbon fiber microelectrode (CFME) modified with nickel oxide and ruthenium hexacyanoferrate (NiO-RuHCF). This sensing layer was deposited electrochemically in a two-step procedure involving an initial galvanostatic NiO deposition followed by a potentiodynamic RuHCF deposition from solutions containing the precursor salts. Several important parameters were examined to characterize and optimize the NiO-RuHCF sensing layer with respect to its current response to AA by using cyclic voltammetry, and scanning electron microscopy-energy dispersive X-ray spectroscopy methods. With the NiO-RuHCF coated CFME, the AA oxidation potential under acidic conditions was shifted to a less positive value for about 0.2 V (E(p) of ca. 0.23 V vs. Ag/AgCl) as compared to a bare CFME, which greatly improves the electrochemical selectivity. Using the hydrodynamic amperometry mode, the current vs. AA concentration in 0.01 M HCl, at a selected operating potential of 0.30 V, was found to be linear over a wide range of 10-1610 μM (n=22, r=0.999) with a calculated limit of detection of 1.0 μM. The measurement repeatability was satisfactory with a relative standard deviation (r.s.d.) ranging from 4% to 5% (n=6), depending on the AA concentration, and with a sensor-to-sensor reproducibility (r.s.d.) of 6.9% at 100 μM AA. The long-term reproducibility, using the same microsensor for 112 consecutive measurements of 20 μM AA over 11 h of periodic probing sets over 4 days, was 16.1% r.s.d., thus showing very good stability at low AA levels and suitability for use over a prolonged period of time. Moreover, using the proposed microsensor, additionally coated with a protective cellulose acetate membrane, the calibration plot obtained in the extremely complex matrix of real undiluted gastric juice was linear from 10 to 520 μM (n=14, r=0.998). These results

  6. Experimental Study of A Novel Piezoelectric Tumor Marker Micro-array Immunosensor

    Institute of Scientific and Technical Information of China (English)

    Bo Zhang; Weiling Fu; Xue Zhang; Qinghai Chen; Shijun Xu; Daihua Tang

    2006-01-01

    A novel 2× 5 model of insert-plug piezoelectric quartz crystal tumor marker micro-array immunosensor constructed with screw clamp apparatus has been developed for quantitative detection of the tumor markers such as alpha-fetoprotein (AFP), carcino-embryonic antigen (CEA), prostate specific antigen (PSA), and human chorionic gonadotropin (hCG) in serum, in which every crystal unit can oscillate independently with the stability of ± 1 hertz (Hz)in air and ± 2 Hz in liquid. These response characteristics of Pz tumor marker micro-array immunosensor such as temperature,time-cost, reproducibility and specificity etc were also investigated. The detection ranges for AFP, CEA, PSA, and hCG obtained by Pz micro-array immunosensor were 20 ng/ml~640 ng/ml, 1.56 ng/ml~50 ng/ml, 1.25 ng/ml~50 ng/ml, and 2.5 mIU/ml~250 mIU/ml respectively with the coefficient of variance (CV) less than 5%. No cross-reactivates with other tumor markers in serum were observed. The results of AFP, CEA, PSA, and hCG obtained by this method from 68 serum samples were in good agreement with those given by chemiluminescence immunoassay with the correlation coefficients of 0.92, 0.90, 0.91, and 0.94 respectively. The Pz immunosensor regenerated by urea solution could be reused for five times without appreciable loss of response activity. Therefore, the proposed insert-plug immunosensor provides a rapid,sensitive, specific, reusable, convenient and reliable alternative for the detection of tumor markers in clinical laboratory.

  7. Wireless Amperometric Neurochemical Monitoring Using an Integrated Telemetry Circuit

    Science.gov (United States)

    Roham, Masoud; Halpern, Jeffrey M.; Martin, Heidi B.; Chiel, Hillel J.

    2015-01-01

    An integrated circuit for wireless real-time monitoring of neurochemical activity in the nervous system is described. The chip is capable of conducting high-resolution amperometric measurements in four settings of the input current. The chip architecture includes a first-order ΔΣ modulator (ΔΣM) and a frequency-shift-keyed (FSK) voltage-controlled oscillator (VCO) operating near 433 MHz. It is fabricated using the AMI 0.5 μm double-poly triple-metal n-well CMOS process, and requires only one off-chip component for operation. Measured dc current resolutions of ~250 fA, ~1.5 pA, ~4.5 pA, and ~17 pA were achieved for input currents in the range of ±5, ±37, ±150, and ±600 nA, respectively. The chip has been interfaced with a diamond-coated, quartz-insulated, microneedle, tungsten electrode, and successfully recorded dopamine concentration levels as low as 0.5 μM wirelessly over a transmission distance of ~0.5 m in flow injection analysis experiments. PMID:18990633

  8. Electrochemical fabrication and amperometric sensor application of graphene sheets

    Science.gov (United States)

    Öztürk, Ayşe; Alanyalıoğlu, Murat

    2016-07-01

    Graphene sheets have been fabricated by applying two-step electrochemical processes in two-electrode cell system containing 0.1 M sodium dodecyl sulfate (SDS). First step is intercalation of SDS into graphite anode electrode and this process has been applied at different intercalation potential values of 1, 3, 5, and 7 V. Second step includes exfoliation of SDS-intercalated graphite electrode in the same medium by acting as cathode. Stable graphene dispersions are obtained after these two electrochemical steps. Characterization of graphene sheets have been carried out using scanning electron microscopy, electron dispersive spectroscopy, fourier transform infrared spectroscopy, UV-Vis. absorption spectroscopy, X-ray diffraction, and cyclic voltammetry techniques. Graphene sheets have been modified onto glassy carbon electrode (GCE) by drop-casting of graphene dispersion. Graphene/GCE having a good electrocatalytic activity has been used for amperometric determination of nitrite in both standard laboratory and real samples. The oxidation current density was linearly proportional to the nitrite concentration in a range between 1 and 250 μM. The sensitivity of the sensor was calculated as 0.843 μAμM-1 cm-2 with a detection limit of 0.24 μM at a signal-to-noise ratio of 3.0.

  9. Development of an immunosensor using oriented immobilized anti-OmpW for sensitive detection of Vibrio cholerae by surface plasmon resonance.

    Science.gov (United States)

    Taheri, Ramezan Ali; Rezayan, Ali Hossein; Rahimi, Fereshteh; Mohammadnejad, Javad; Kamali, Mehdi

    2016-12-15

    The first SPR sensor for detection of bacteria was reported in 1998 with high detection limit as much as 10(7)cfu/mL. Since then, a lot of effort has been made to lower detection limit and increase sensitivity of detection mainly by using of different assay formats, immobilization strategies, suitable antibodies, minimizing non-specific adsorption and improving the quality of SPR devices. The aim of this paper is to introduce the potential of an antibody against recombinant outer membrane protein (anti-OmpW) in sensitive detection of Vibrio cholerae by developing an immunosensor based on SPR and compare the sensitivity of this method with former report for detection of V. cholerae published in 2006. Recombinant OmpW antigen (a bacterial outer-membrane protein) of V. cholerae was expressed and purified and raising of polyclonal rabbit anti-OmpW was done. Protein G was covalently immobilized on 11-MUA SAM via amine coupling and bioaffinity-based oriented immobilization of anti-OmpW was done on protein G layer. The results showed high affinity interaction between OmpW and anti-OmpW (KD=2.4×10(-9)M) and the detection limit of fabricated immunosensor was 43 cells/mL. The apparent reasons for achieving this low LOD are discussed.

  10. Open Tubular Microreactor with Enzyme Functionalized Micro- fluidic Channel for Amperometric Detection of Glucose

    Institute of Scientific and Technical Information of China (English)

    张蕾; 曲平; 盛金; 雷建平; 鞠烷先

    2012-01-01

    A simple and efficient method using enzyme immobilized microfluidic channel as open tubular microreactor was designed for amperometric detection of glucose. The microreactor was composed of a polydimethylsilicone/ glass hybrid device with three reservoirs, a cooling cave and a 6 cm capillary with a sampling fracture as micro-channel. The microchannel was further modified by thermal polymerization, followed by covalently attaching with glucose oxidase. Through fracture sampling and electrochromatography separation, the production via enzymatic reaction was determinated by Pt electrode at the end of capillary. The linear range for the detection of glucose was 0.05--7.5 mmol·L-1 with detection limit of 23μmol.L-1 The inter-and intra-chip reproducibilities for determination of 2.5 mmol-L-1 glucose were 98.5% (n=5) and 96.0% (n=5), respectively. With the advantage of flexible assembly, rapid efficiency, good stability and low-cost, this microreactor provided a potential platform for estab- lishing a portable enzyme-based chemical detection system in practical application.

  11. Determination of Four Active Ingredients in Vc Yinqiao Tablets by Capillary Zone Electrophoresis with Amperometric Detection

    Institute of Scientific and Technical Information of China (English)

    L(U),Jin; WANG,Qing-Jiang; CHENG,Xi; LIU,Hai-Yan; HE,Pin-Gang; FANG,Yu-Zhi

    2006-01-01

    A simple, reliable and reproducible method, based on capillary zone electrophoresis with amperometric detection (CZE-AD), has been developed for simultaneous determination of four active ingredients in Vc Yinqiao tablets including paracetamol, vitamin C, caffeic acid and chlorogenic acid. A carbon-disk electrode was used as working electrode and 0.95 V (versus SCE) was selected as detection potential. The optimal conditions of CZE experiment were 30 mmol·L-1 borate solution (pH 9.5) as running buffer, 14 kV as separation voltage and 8 s (14 kV) as electro-kinetic sampling time. Under the selected optimum conditions, paracetamol, vitamin C, caffeic acid and chlorogenic acid could be perfectly separated within 22 min, and their detection limits (S/N=3) ranged from 5 × 10-7 to 1×10-6 mol·L-1. This proposed method demonstrated good reproducibility with relative standard deviations of less than 3% for both migration time and peak current (n=7). The utility of this method was demonstrated by monitoring a kind of compound medicine named Vc Yinqiao tablets and the assay results were satisfactory.

  12. Assessment of the uncertainty budget for the amperometric measurement of dissolved oxygen.

    Science.gov (United States)

    Fisicaro, Paola; Adriaens, Annemie; Ferrara, Enzo; Prenesti, Enrico

    2007-07-30

    This work aimed at identifying the main sources of uncertainty for the measurement of dissolved oxygen concentration in aqueous solutions. The experimental apparatus consists of an amperometric cell based on the Clark-type sensor. The corresponding uncertainty budget was assessed, this being a fundamental step for the validation of a measurement method. The principle of the measurement, as well as the procedure for the set-up and the characterisation of the cell, are described. The measurement equation was defined as a combination of Faraday's and Fick's laws, and a method was worked out for the empirical determination of the diffusivity parameter. In this connection, the solutions of oxygen were standardised by way of the Winkler's titration, as suggested by the ISO Guide 5813 and 5814. With this approach we aimed at contributing to the development of a potential primary method of measurement. A discussion of all the contributions to the overall uncertainty is reported, allowing operators to locate the largest ones and plan specific improvements.

  13. Simultaneous Voltammetric/Amperometric Determination of Sulfide and Nitrite in Water at BDD Electrode

    Directory of Open Access Journals (Sweden)

    Anamaria Baciu

    2015-06-01

    Full Text Available This work reported new voltammetric/amperometric-based protocols using a commercial boron-doped diamond (BDD electrode for simple and fast simultaneous detection of sulfide and nitrite from water. Square-wave voltammetry operated under the optimized working conditions of 0.01 V step potential, 0.5 V modulation amplitude and 10 Hz frequency allowed achieving the best electroanalytical parameters for the simultaneous detection of nitrite and sulfide. For practical in-field detection applications, the multiple-pulsed amperometry technique was operated under optimized conditions, i.e., −0.5 V/SCE for a duration of 0.3 s as conditioning step, +0.85 V/SCE for a duration of 3 s that assure the sulfide oxidation and +1.25 V/SCE for a duration of 0.3 s, where the nitrite oxidation occurred, which allowed the simultaneously detection of sulfide and nitrite without interference between them. Good accuracy was found for this protocol in comparison with standardized methods for each anion. Also, no interference effect was found for the cation and anion species, which are common in the water matrix.

  14. Simultaneous Voltammetric/Amperometric Determination of Sulfide and Nitrite in Water at BDD Electrode

    Science.gov (United States)

    Baciu, Anamaria; Ardelean, Magdalena; Pop, Aniela; Pode, Rodica; Manea, Florica

    2015-01-01

    This work reported new voltammetric/amperometric-based protocols using a commercial boron-doped diamond (BDD) electrode for simple and fast simultaneous detection of sulfide and nitrite from water. Square-wave voltammetry operated under the optimized working conditions of 0.01 V step potential, 0.5 V modulation amplitude and 10 Hz frequency allowed achieving the best electroanalytical parameters for the simultaneous detection of nitrite and sulfide. For practical in-field detection applications, the multiple-pulsed amperometry technique was operated under optimized conditions, i.e., −0.5 V/SCE for a duration of 0.3 s as conditioning step, +0.85 V/SCE for a duration of 3 s that assure the sulfide oxidation and +1.25 V/SCE for a duration of 0.3 s, where the nitrite oxidation occurred, which allowed the simultaneously detection of sulfide and nitrite without interference between them. Good accuracy was found for this protocol in comparison with standardized methods for each anion. Also, no interference effect was found for the cation and anion species, which are common in the water matrix. PMID:26102487

  15. Amperometric Choline Biosensor Fabricated through Electrostatic Assembly of Bienzyme/Polyelectrolyte Hybrid Layers on Carbon Nanotubes

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Jun; Liu, Guodong; Lin, Yuehe

    2006-03-01

    We report a flow injection amperometric choline biosensors based on the electrostatic assembly of an enzyme of choline oxidase (ChO) and a bi-enzyme of ChO and horseradish peroxidase (HRP) onto multi-wall carbon nanotubes (MWCNT) modified glassy carbon (GC) electrodes. These choline biosensors were fabricated by immobilization of enzymes on the negatively charged MWCNT surface through alternatively assembling a cationic polydiallydiimethylammonium chloride (PDDA) layer and an enzyme layer. Using this layer-by-layer assembling approach, bioactive nanocomposite film of a PDDA/ChO/PDDA/HRP/PDDA/CNT (ChO/HRP/CNT) and a PDDA/ChO/PDDA/ CNT (ChO/ CNT) were fabricated on GC surface, respectively. Owning to the electrocatalytic effect of carbon nanotubes, the measurement of faradic responses resulting from enzymatic reactions has been realized at low potential with acceptable sensitivity. It is found the ChO/HRP/CNT biosensor is more sensitive than the ChO/CNT one. Experimental parameters affecting the sensitivity of biosensors, e.g. applied potential, flow rate, etc. were optimized and potential interference was examined. The response time for this choline biosensor is fast (less than a few seconds). The linear range of detection for the choline biosensor is from 5 x 10-5 to 5 x 10-3 M and the detection limit is determined to be about 1.0 x 10-5 M.

  16. Fast determination of aldehyde preservatives by miniaturized capillary electrophoresis with amperometric detection.

    Science.gov (United States)

    Li, Ying; Chen, Fang; Ge, Jinyuan; Tong, Fanghong; Deng, Zhaoyue; Shen, Fengwu; Gu, Qianxia; Ye, Jiannong; Chu, Qingcui

    2014-02-01

    A novel miniaturized CE with amperometric detection (mini-CE-AD) method has been developed for fast determination of aliphatic aldehyde preservatives, namely formaldehyde and glyoxal, in commodities. After derivatization with an electroactive compound 2-thiobarbituric acid, these two nonelectroactive aldehydes were converted to electroactive adducts, therefore detectable by mini-CE-AD approach. Under the optimum conditions, two aldehydes can be well-separated with the coexisting interferents as well as their homologs (acetaldehyde and methyl-glyoxal), and the LODs (S/N = 3) were achieved at nanogram-per-milliliter level (1.64-2.80 ng/mL) based on the online enrichment method of transient moving chemical reaction boundary. The proposed method has been applied for the analyses of above aldehyde preservatives in different real commodity samples including skincare products, baby lotion, and toothpaste, and the average recoveries were in the range of 94-105%, which should find a wide range of analytical applications as an alternative to conventional and microchip CE approaches.

  17. Ceramic Carbon/Polypyrrole Materials for the Construction of Bienzymatic Amperometric Biosensor for Glucose

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    A novel amperometric glucose biosensor was constructed by electrochemical formation of a polypyrrole (PPy) membrane in the presence of glucose oxidase (GOD) on the surface of a horseradish peroxidase (HRP) modified ferrocenecarboxylic acid (FCA) mediated sol-gel derived ceramic carbon electrode. The amperometric detection of glucose was carried out at +0.16 V (vs. SCE) in 0.1 mol/L phosphate buffer solution (pH 6.9) with a linear response range between 8.0×10-5 and 1.3×10-3 mol/L of glucose. The biosensor showed a good suppression of inter- ference and a negligible deviation in the amperometric detection.

  18. A Label-Free Impedance Immunosensor Using Screen-Printed Interdigitated Electrodes and Magnetic Nanobeads for the Detection of E. coli O157:H7.

    Science.gov (United States)

    Wang, Ronghui; Lum, Jacob; Callaway, Zach; Lin, Jianhan; Bottje, Walter; Li, Yanbin

    2015-12-15

    Escherichia coli O157:H7 is one of the leading bacterial pathogens causing foodborne illness. In this study, an impedance immunosensor based on the use of magnetic nanobeads and screen-printed interdigitated electrodes was developed for the rapid detection of E. coli O157:H7. Magnetic nanobeads coated with anti-E. coli antibody were mixed with an E. coli sample and used to isolate and concentrate the bacterial cells. The sample was suspended in redox probe solution and placed onto a screen-printed interdigitated electrode. A magnetic field was applied to concentrate the cells on the surface of the electrode and the impedance was measured. The impedance immunosensor could detect E. coli O157:H7 at a concentration of 10(4.45) cfu·mL(-1) (~1400 bacterial cells in the applied volume of 25 μL) in less than 1 h without pre-enrichment. A linear relationship between bacteria concentration and impedance value was obtained between 10(4.45) cfu·mL(-1) and 10(7) cfu·mL(-1). Though impedance measurement was carried out in the presence of a redox probe, analysis of the equivalent circuit model showed that the impedance change was primarily due to two elements: Double layer capacitance and resistance due to electrode surface roughness. The magnetic field and impedance were simulated using COMSOL Multiphysics software.

  19. Study on the immobilization of anti-IgG on Au-colloid modified gold electrode via potentiometric immunosensor, cyclic voltammetry, and electrochemical impedance techniques.

    Science.gov (United States)

    Fu, Yingzi; Yuan, Ruo; Tang, Dianping; Chai, Yaqin; Xu, Lan

    2005-01-15

    The immobilization of anti-IgG on Au-colloid modified gold electrodes has been investigated. A cleaned gold electrode was first immersed in a mercaptoethylamine (AET) solution, and then gold nanoparticles were chemisorbed onto the thiol groups of the mercaptoethylamine. Finally, anti-IgG was adsorbed onto the surface of the gold nanoparticles. Potentiometric immunosensor, cyclic voltammetry, and electrochemical impedance techniques were used to investigate the immobilization of anti-IgG on Au colloids. In the impedance spectroscopic study, an obvious difference of the electron transfer resistance between the Au-colloid modified electrode and the bare gold electrode was observed. The cyclic voltammogram tends to be more irreversible with increased anti-IgG concentration. Using the potentiometric immunosensor, the proposed technique is based on that the specific agglutination of antibody-coated gold nanoparticles, averaging 16 nm in diameter, in the presence of the corresponding antigen causes a potential change that is monitored by a potentiometry. It is found that the developed immunoagglutination assay system is sensitive to the concentration of IgG antigen as low as 12 ng mL(-1). Experimental results showed that the developed technique is in satisfactory agreement with the ELISA method, and that gold nanoparticles can be used as a biocompatible matrix for antibody or antigen immobilization.

  20. Demonstration of submersible high-throughput microfluidic immunosensors for underwater explosives detection.

    Science.gov (United States)

    Adams, André A; Charles, Paul T; Deschamps, Jeffrey R; Kusterbeck, Anne W

    2011-11-15

    Significant security threats posed by highly energetic nitroaromatic compounds in aquatic environments and the demilitarization and pending cleanup of areas previously used for munitions manufacture and storage represent a challenge for less expensive, faster, and more sensitive systems capable of analyzing groundwater and seawater samples for trace levels of explosive materials. Presented here is an inexpensive high throughput microfluidic immunosensor (HTMI) platform intended for the rapid, highly selective quantitation of nitroaromatic compounds in the field. Immunoaffinity and fluorescence detection schemes were implemented in tandem on a novel microfluidic device containing 39 parallel microchannels that were 500 μm tall, 250 μm wide, and 2.54 cm long with covalently tethered antibodies that was engineered for high-throughput high-volume sample processing. The devices were produced via a combination of high precision micromilling and hot embossing. Mass transfer limitations were found in conventional microsystems and were minimized due to higher surface area to volume ratios that exceeded those possessed by conventional microdevices and capillaries. Until now, these assays were limited to maximum total volume flow rates of ~1 mL/min due in part to kinetics and high head pressures of single microchannels. In the design demonstrated here, highly parallelized microchannels afforded up to a 100-fold increase in total volume flow rate while maintaining favorable kinetic constraints for efficient antigen-antibody interaction. The assay employed total volume throughput of up to 6 mL/min while yielding signal-to-noise ratios of >15 in all cases. In addition to samples being processed up to 60 times faster than in conventional displacement-based immunoassays, the current system was capable of quantitating 0.01 ng/mL TNT samples without implementing offline preconcentration, thereby, demonstrating the ability to improve sensitivity by as much as 2 orders of magnitude

  1. Fabrication of multiwalled carbon nanotube-polyaniline/platinum nanocomposite films toward improved performance for a cholesterol amperometric biosensor.

    Science.gov (United States)

    Xu, ZeHong; Cheng, XiaoDan; Tan, JianHong; Gan, Xianxue

    2016-11-01

    A simple and high sensitive cholesterol amperometric biosensor, which is based on in situ electropolymerization of multi-walled carbon nanotube-polyaniline (MWCNT-PANI) nanocomposite and electrodeposition of platinum nanoparticle (nano-Pt) films onto the glassy carbon electrode surface for cholesterol oxidase immobilization, was constructed in this study. The preparation process of the modified electrode was characterized by cyclic voltammetry, electrochemical impedance spectroscopy, scanning electron microscopy, and chronoamperometry. Because of the synergistic electrocatalytic activity between MWCNT-PANI nanocomposites and nano-Pt, the cholesterol biosensor exhibited an excellent performance with a linear range of 2.0-510.0 µM, a detection limit of 0.8 µM (signal-to-noise ratio = 3), a high sensitivity of 109.9 µA mM(-1) , and a short response time within 5 Sec. Moreover, the reproducibility, stability, and selectivity of the biosensor were also investigated.

  2. Amperometric detection of heavy metal ions in ion pair chromatography at an array of water/nitrobenzene micro interfaces

    Energy Technology Data Exchange (ETDEWEB)

    Wilke, S. [Martin-Luther-Univ. Halle-Wittenberg, Merseburg (Germany). Fachbereich Chemie; Wang, H. [Martin-Luther-Univ. Halle-Wittenberg, Merseburg (Germany). Fachbereich Chemie; Muraczewska, M. [Martin-Luther-Univ. Halle-Wittenberg, Merseburg (Germany). Fachbereich Chemie; Mueller, H. [Martin-Luther-Univ. Halle-Wittenberg, Merseburg (Germany). Fachbereich Chemie

    1996-10-01

    A novel amperometric detector for heavy metal ions has been developed and successfully applied for ion pair chromatography. The detector is based on the electrochemical transfer of the metal ions across an array of water/nitrobenzene micro interfaces. The ion transfer is facilitated by the neutral ionophores methylenebis(diphenylphosphineoxide) and methylenebis(di- phenylphosphinesulfide). More than eight metals are separated in less than 15 min on an RP18 column using octyl sulfonate as ion pair reagent. For the heavy metals, the limits of decision are 19(Pb{sup 2+}), 9(Zn{sup 2+}), 9l (Co{sup 2+}), 8(Cd{sup 2+}) and 1.6(Mn{sup 2+}) {mu}g/L. The applicability of the new method for water samples is demonstrated. (orig.). With 3 figs., 2 tabs.

  3. Development of a novel, sensitive amperometric-FIA glucose biosensor by packing up the amperometric cell with glucose oxidase modified anion exchange resin.

    Science.gov (United States)

    Su, Yuhua; Huang, Weixiong; Hu, Rongzong; Ding, Haodong; Hu, Kangkang

    2009-04-15

    In this work, the anion exchange resin (AER) was modified with a layer of glucose oxidase (GOD) and poly(diallyldimethylammonium chloride) (PDDA), respectively, via layer-by-layer electrostatic self-assembling strategy. The PDDA and GOD modified AER (PDDA/GOD/AER) was then packed into a home-made amperometric cell for flow injection analysis (FIA) of glucose. This design simplified the setup by integrating the enzyme reactor into the amperometric cell. And the AER in the cell behaved bifunctional, it was not only the support of enzymes, but also an anti-interference tool due to its retention effect toward ascorbic acid (AA) and uric acid (UA). A platinum modified porous titanium (Pt/PTi) electrode was utilized in the cell as the working electrode (WE), due to its large effective surface area it could increase the response by 8.3 times as compared with the planar pure platinum electrode. The proposed biosensor was very sensitive (22.4 microA cm(-2) mM(-1)) in glucose quantification, and the linear range was from 1 micromol L(-1) to 2 mmol L(-1) with the detection limit of 0.8 micromol L(-1). The biosensor was used for serum glucose determination, and the result obtained was satisfying. This work may have provided a reference design of the amperometric cell which could be adopted in other enzymatic-FIA biosensors.

  4. Sensitive voltammetric and amperometric responses of respiratory toxins at hemin-adsorbed carbon-felt

    Institute of Scientific and Technical Information of China (English)

    Yasushi Hasebe; Yue Wang

    2013-01-01

    A hemin [iron-Fe(Ⅲ) protoporphyrin Ⅸ chloride] was adsorbed onto a carbon-felt (CF),which is a microelectrode ensemble of micro carbon fiber (ca.7 μm diameter).The resulting hemin-adsorbed-CF (hemin-CF) showed a well-defined redox wave based on the hemin-Fe(2)/Fe(Ⅱ) redox process with the formal potential of-0.225 Ⅴ vs.Ag/AgCI in deoxygenated phosphate/citrate buffer solution (0.1mol/L,pH 5.0).The apparent heterogenous electron transfer rate constant was estimated to be 8.6 sec-1.In air-saturated electrolyte solution,the hemin-CF exhibited an excellent electrocatalytic activity for the reduction of dioxygen (O2).This activity was reversibly inhibited by respiratory toxins such as cyanide and azide,which bind sixth coordination position of iron active center of hemin.The electrocatalytic O2 reduction current at the hemin-CF was modulated by the toxins in a concentration-depending manner.Based on the relationship between the %inhibition and the toxin concentration,apparent inhibition constants of cyanide and azide were evaluated to be 4.52 and 1.98 μmol/L,respectively.When the hemin-CF was used as a working electrode unit of the CF-based electrochemical flow-through detector with air-saturated carrier,the injection of the azide induced peak-shape current responses,which allowed rapid and continuous flow-amperometric determination of azide with high sensitivity.

  5. Electrochemical magneto immunosensor for the detection of anti-TG2 antibody in celiac disease.

    Science.gov (United States)

    Kergaravat, Silvina V; Beltramino, Luis; Garnero, Nidia; Trotta, Liliana; Wagener, Marta; Isabel Pividori, Maria; Hernandez, Silvia R

    2013-10-15

    An electrochemical magneto immunosensor for the detection of anti-transglutaminase antibodies (ATG2) in celiac disease was developed. The immunological reaction is performed on magnetic beads (MBs) as a solid support in which the transglutaminase enzyme (TG2) is covalently immobilized (TG2-MB) and then ATG2 were revealed by an antibody labeled with peroxidase. The electrochemical response of the enzymatic reaction with o-phenilendiamine and H₂O₂ as substrates by square wave voltammetry was correlated with the ATG2. Graphite-epoxi composite cylindrical electrodes and screen printed electrodes were used as transducers in the immunosensor. A total number of 29 sera from clinically confirmed cases of celiac disease and 19 negative control sera were tested by the electrochemical magneto immunosensor. The data were submitted to the receiver-operating characteristic plot (ROC) analysis which indicated that 16.95 units was the most effective cut-off value (COV) to discriminate correctly between celiac and non-celiac patients. Using this point for prediction, sensitivity was found to be 100%, while specificity was 84%.

  6. A disposable electrochemical immunosensor for prolactin involving affinity reaction on streptavidin-functionalized magnetic particles

    Energy Technology Data Exchange (ETDEWEB)

    Moreno-Guzman, Maria; Gonzalez-Cortes, Araceli [Department of Analytical Chemistry, Faculty of Chemistry, University Computense of Madrid, 28040 Madrid (Spain); Yanez-Sedeno, Paloma, E-mail: yseo@quim.ucm.es [Department of Analytical Chemistry, Faculty of Chemistry, University Computense of Madrid, 28040 Madrid (Spain); Pingarron, Jose M. [Department of Analytical Chemistry, Faculty of Chemistry, University Computense of Madrid, 28040 Madrid (Spain)

    2011-04-29

    A novel electrochemical immunosensor was developed for the determination of the hormone prolactin. The design involved the use of screen-printed carbon electrodes and streptavidin-functionalized magnetic particles. Biotinylated anti-prolactin antibodies were immobilized onto the functionalized magnetic particles and a sandwich-type immunoassay involving prolactin and anti-prolactin antibody labelled with alkaline phosphatase was employed. The resulting bio-conjugate was trapped on the surface of the screen-printed electrode with a small magnet and prolactin quantification was accomplished by differential pulse voltammetry of 1-naphtol formed in the enzyme reaction using 1-naphtyl phosphate as alkaline phosphatase substrate. All variables involved in the preparation of the immunosensor and in the electrochemical detection step were optimized. The calibration plot for prolactin exhibited a linear range between 10 and 2000 ng mL{sup -1} with a slope value of 7.0 nA mL ng{sup -1}. The limit of detection was 3.74 ng mL{sup -1}. Furthermore, the modified magnetic beads-antiprolactin conjugates showed an excellent stability. The immunosensor exhibited also a high selectivity with respect to other hormones. The analytical usefulness of the immnunosensor was demonstrated by analyzing human sera spiked with prolactin at three different concentration levels.

  7. Optical immunosensors for detection of Listeria monocytogenes and Salmonella enteritidis from food

    Science.gov (United States)

    Bhunia, Arun K.; Geng, Tao; Lathrop, Amanda; Valadez, Angela; Morgan, Mark T.

    2004-03-01

    Listeria monocytogenes and Salmonella are two major foodborne pathogens of significant concern. Two optical evanescent wave immunosensors were evaluated for detection: Antibody-coupled fiber-optic biosensor and a surface plasmon resonant (SPR) immunosensor. In the fiber-optic sensor, polyclonal antibodies for the test organisms were immobilized on polystyrene fiber wave -guides using streptavidin - biotin chemistry. Cyanine 5 -labeled monoclonal antibodies C11E9 (for L. monocytogenes) and SF-11 (for Salmonella Enteritidis) were used to generate a specific fluorescent signal. Signal acquisition was performed by launching a laser-light (635 nm) from an Analyte-2000. This immunosensor was able to detect 103 - 109 cfu/ml of L. monocytogenes or 106-109 cfu/ml of Salmonella Enteritidis and the assays were conducted at near real-time with results obtained within one hour of sampling. The assays were specific and showed signal even in the presence of other microorganisms such as E. coli, Enterococcus faecalis or Salmonella Typhimurium. In the SPR system, IAsys instrument (resonant mirror sensor) was used. Monoclonal antibody-C11E9 was directly immobilized onto a carboxylate cuvette. Whole Listeria cells at various concentrations did not yield any signal while surface protein extracts did. Crude protein extracts from L. monocytogenes and L. innocua had average binding responses of around 150 arc sec (0.25 ng/mm2), which was significantly different from L. grayi, L. ivanovii, or L. welshimeri with average responses of Salmonella Enteritidis.

  8. A Doped Polyaniline Modified Electrode Amperometric Biosensor for Gluconic Acid Determination in Grapes

    Directory of Open Access Journals (Sweden)

    Donatella Albanese

    2014-06-01

    Full Text Available In winemaking gluconic acid is an important marker for quantitative evaluation of grape infection by Botrytis cinerea. A screen-printed amperometric bienzymatic sensor for the determination of gluconic acid based on gluconate kinase (GK and 6-phospho-D-gluconate dehydrogenase (6PGDH coimmobilized onto polyaniline/poly (2-acrylamido-2-methyl-1-propanesulfonic acid; PANI-PAAMPSA is reported in this study. The conductive polymer electrodeposed on the working electrode surface allowed the detection of NADH at low potential (0.1 V with a linear range from 4 × 10−3 to 1 mM (R2 = 0.99 and a sensitivity of 419.44 nA∙mM−1. The bienzymatic sensor has been optimized with regard to GK/6PGDH enzymatic unit ratio and ATP/NADP+ molar ratio which resulted equal to 0.33 and 1.2, respectively. Under these conditions a sensitivity of 255.2 nA∙mM−1, a limit of detection of 5 μM and a Relative Standard Deviation (RSD of 4.2% (n = 5 have been observed. Finally, the biosensor has been applied for gluconic acid measurements in must grape samples and the matrix effect has been taken into consideration. The results have been compared with those obtained on the same samples with a commercial kit based on a spectrophotometric enzyme assay and were in good agreement, showing the capability of the bienzymatic PANI-PAAMPSA biosensor for gluconic acid measurements and thus for the evaluation of Botrytis cinerea infection in grapes.

  9. Biossensores amperométricos para determinação de compostos fenólicos em amostras de interesse ambiental Amperometric biosensors for phenolic compounds determination in the environmental interess samples

    Directory of Open Access Journals (Sweden)

    Simone Soares Rosatto

    2001-02-01

    Full Text Available Phenols are widely used in many areas and commonly found as industrial by-products. A great number of agricultural and industrial activities realise phenolic compounds in the environmental. Waste phenols are produced mainly by the wood-pulp industry and during production of synthetic polymers, drugs, plastics, dyes, pesticides and others. Phenols are also released into the environmental by the degradation of pesticides with phenolic skeleton. The phenols level control is very important for the environmental protection. Amperometric biosensor has shown the feasibility to complement laboratory-based analytical methods for the determination of phenolic compounds, providing alternatives to conventional methods which have many disadvantages. This brief review considers the evolution of an approach to amperometric measurement using the catalytic properties of some enzymes for phenolic compounds monitoring.

  10. Extended-gate-type IGZO electric-double-layer TFT immunosensor with high sensitivity and low operation voltage

    Science.gov (United States)

    Liang, Lingyan; Zhang, Shengnan; Wu, Weihua; Zhu, Liqiang; Xiao, Hui; Liu, Yanghui; Zhang, Hongliang; Javaid, Kashif; Cao, Hongtao

    2016-10-01

    An immunosensor is proposed based on the indium-gallium-zinc-oxide (IGZO) electric-double-layer thin-film transistor (EDL TFT) with a separating extended gate. The IGZO EDL TFT has a field-effect mobility of 24.5 cm2 V-1 s-1 and an operation voltage less than 1.5 V. The sensors exhibit the linear current response to label-free target immune molecule in the concentrations ranging from 1.6 to 368 × 10-15 g/ml with a detection limit of 1.6 × 10-15 g/ml (0.01 fM) under an ultralow operation voltage of 0.5 V. The IGZO TFT component demonstrates a consecutive assay stability and recyclability due to the unique structure with the separating extended gate. With the excellent electrical properties and the potential for plug-in-card-type multifunctional sensing, extended-gate-type IGZO EDL TFTs can be promising candidates for the development of a label-free biosensor for public health applications.

  11. A Facile Electrochemical Preparation of Reduced Graphene Oxide@Polydopamine Composite: A Novel Electrochemical Sensing Platform for Amperometric Detection of Chlorpromazine

    Science.gov (United States)

    Palanisamy, Selvakumar; Thirumalraj, Balamurugan; Chen, Shen-Ming; Wang, Yi-Ting; Velusamy, Vijayalakshmi; Ramaraj, Sayee Kannan

    2016-01-01

    We report a novel and sensitive amperometric sensor for chlorpromazine (CPZ) based on reduced graphene oxide (RGO) and polydopamine (PDA) composite modified glassy carbon electrode. The RGO@PDA composite was prepared by electrochemical reduction of graphene oxide (GO) with PDA. The RGO@PDA composite modified electrode shows an excellent electro-oxidation behavior to CPZ when compared with other modified electrodes such as GO, RGO and GO@PDA. Amperometric i-t method was used for the determination of CPZ. Amperometry result shows that the RGO@PDA composite detects CPZ in a linear range from 0.03 to 967.6 μM. The sensor exhibits a low detection limit of 0.0018 μM with the analytical sensitivity of 3.63 ± 0.3 μAμM–1 cm–2. The RGO@PDA composite shows its high selectivity towards CPZ in the presence of potentially interfering drugs such as metronidazole, phenobarbital, chlorpheniramine maleate, pyridoxine and riboflavin. In addition, the fabricated RGO@PDA modified electrode showed an appropriate recovery towards CPZ in the pharmaceutical tablets. PMID:27650697

  12. Microchip Capillary Electrophoresis with an End-Channel Amperometric Detector and Its Preliminary Application

    Institute of Scientific and Technical Information of China (English)

    吴友谊; 屈锋; 林金明

    2005-01-01

    An end-channel amperometric detector with a guide tube for working electrode was designed and integrated on a home-made glass microchip. The guide tube was directly patterned and fabricated at the end of the detection reservoir, which made the fixation and alignment of working electrode relatively easy. The fabrication was carried out in a two-step etching process. A 30 μm carbon fiber microdisk electrode and Pt cathode were also integrated onto the amperometric detector. The baseline separation of dopamine (DA), catechol (CA) and epinephrine (EP) was achieved within 80 s. Relative standard deviations of not more than 5.2% were obtained for both peak currents and migration times of DA and CA (n=5). Using standard adding method, DA in tLrine and plasma samples was detected. The recoveries were in the range of 83%—103%.

  13. A reagentless enzymatic amperometric biosensor using vertically aligned carbon nanofibers (VACNF)

    Energy Technology Data Exchange (ETDEWEB)

    Weeks, Martha L [University of Tennessee, Knoxville (UTK); Rahman, Touhidur [ORNL; Frymier, Paul Dexter [ORNL; Islam, Syed K [University of Tennessee, Knoxville (UTK); McKnight, Timothy E [ORNL

    2008-01-01

    A reagentless amperometric enzymatic biosensor is constructed on a carbon substrate for detection of ethanol. Yeast alcohol dehydrogenase (YADH), an oxidoreductase, and its cofactor nicotinamide adenine dinucleotide (NAD+) are immobilized by adsorption and covalent attachment to the carbon substrate. Carbon nanofibers grown by plasma enhanced chemical vapor deposition (PECVD) are chosen as the electrode material due to their excellent structural and electrical properties. Electrochemical techniques are employed to test the functionality and performance of the biosensor using reduced form of nicotinamide adenine dinucleotide (NADH) which also determines the oxidation peak potential of NADH. Subsequently, amperometric measurements are conducted for detection of ethanol to determine the electrical current response due to the increase in analyte concentration. The detection range, storage stability, reusability, and response time of the biosensor are also examined.

  14. Determination of catecholamines in plasma by HPLC and amperometric detection. Comparison with a radioenzymatic method.

    Science.gov (United States)

    Bauersfeld, W; Ratge, D; Knoll, E; Wisser, H

    1986-03-01

    The determination of norepinephrine and epinephrine in plasma by HPLC with amperometric detection was modified, giving detection limits of 25 ng/l for norepinephrine and epinephrine, respectively, using 1 ml plasma. In order to achieve this sensitivity, it was necessary to minimize the background noise by modification of instrumentation and specimen handling. Particularly important was the extra purification of the reagents, the application of micro-bore HPLC, the enzymatic cleavage of uric acid and temperature control of the amperometric cell and the amplifier. Comparison of the present method with the radioenzymatic determination of catecholamines resulted in coefficients of correlation of r = 0.924 and 0.919 for norepinephrine and epinephrine, resp. (n = 38). The concentrations of the 38 different samples used for the comparison were in the physiological range.

  15. Construction and evaluation of a novel end-column amperometric detection system for electrophoresis microchips

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    A set of integrated end-column amperometric detection system has been developed,onto which an electrophoresis microchip can be conveniently integrated.Finely machined by a piece of transparent organic glass,the system consists of an electrophoresis microchip platform and an amperometric detection reservoir,in which the microchip can be fixed onto the platform by microchip grooves and with stainless steel fixture.Each detection electrode can be directly fixed in the amperometric detection reservoir by screws and nuts.With dopamine as the model analyte,we take platinum disc electrodes with different diameters of 100 μm,300 μm and 500 μm and a carbon fiber electrode with the diameter of 240 μm as the working electrode,all of which accomplish sensitive detection.The detection parameters of the system are optimized with the carbon fiber electrode.The detection results show that in the electrochemical cleaning procedure,the relative deviations of 3.2% and 0.5% for the peak current and retention time,respectively,can be obtained for the successive detections of 100 μM dopamine,and the limit of detection for dopamine can reach 0.4 μM(S/N = 3).This system is small,but its performance of detection is stable and sensitive,and the replacement of its working electrodes is convenient,so it is very suitable as a universal platform of end-column amperometric detection for electrophoresis microchips.

  16. Amperometric Determination of Sulfite by Gas Diffusion-Sequential Injection with Boron-Doped Diamond Electrode

    OpenAIRE

    Orawon Chailapakul; Toshihiko Imato; Narong Praphairaksit; Kulwadee Pinwattana; Chakorn Chinvongamorn

    2008-01-01

    A gas diffusion sequential injection system with amperometric detection using aboron-doped diamond electrode was developed for the determination of sulfite. A gasdiffusion unit (GDU) was used to prevent interference from sample matrices for theelectrochemical measurement. The sample was mixed with an acid solution to generategaseous sulfur dioxide prior to its passage through the donor channel of the GDU. Thesulfur dioxide diffused through the PTFE hydrophobic membrane into a carrier solution...

  17. Amperometric post spike feet reveal most exocytosis is via extended kiss-and-run fusion

    OpenAIRE

    Lisa J Mellander; Raphaël Trouillon; Svensson, Maria I.; Ewing, Andrew G.

    2012-01-01

    The basis for communication between nerve cells lies in the process of exocytosis, the fusion of neurotransmitter filled vesicles with the cell membrane resulting in release of the signaling molecules. Even though much is known about this process, the extent that the vesicles are emptied upon fusion is a topic that is being debated. We have analyzed amperometric peaks corresponding to release at PC12 cells and find stable plateau currents during the decay of peaks, indicating closing of the v...

  18. A Fully Automated Sequential-Injection Analyser for Dual Electrogenerated Chemiluminescence/Amperometric Detection

    OpenAIRE

    Economou, Anastasios; Nika, Maria

    2006-01-01

    This work describes the development of a dedicated, fully automated sequential-injection analysis (SIA) apparatus suitable for simultaneous electrogenerated chemiluminescence (ECL) and amperometric detection. The instrument is composed of a peristaltic pump, a multiposition selection valve, a home-made potentiostat, a thin-layer electrochemical/optical flow-through cell, and a light detector. Control of the experimental sequence and simultaneous data acquisition of the light and the current i...

  19. A New Amperometric Glucose Biosensor with Naphthol Green B as Mediator

    Institute of Scientific and Technical Information of China (English)

    Qin ZHAO; Ruo YUAN; Chang Li MO; Ya Qin CHAI; Xia ZHONG

    2004-01-01

    Naphthol green B was used, for the first time, as a new mediator in an amperometric glucose biosensor. It is a good mediator, promoting electron transfer from glucose oxidase to graphite electrode. The biosensor shows high sensitivity to glucose at low potential with response time of 30 seconds. The linear range is from 1.5 to 18 μmol/L glucose with detection limit of 0.5 μmol/L glucose.

  20. Anodic Voltammetry of Thioacetamide and its Amperometric Determination in Aqueous Media

    Directory of Open Access Journals (Sweden)

    Daniela Dascălu

    2008-08-01

    Full Text Available TAA is a harmful, presumptive pollutant in tap waters and waste waters. Several alternatives have been tested as new possibilities for the anodic determination of TAA in aqueous solutions, simulated waste waters and tap water. The electrochemical behaviour of thioacetamide (TAA was investigated at a boron-doped diamond (BDD electrode both in unbuffered 0.1 M Na2SO4 and buffered solutions as supporting electrolytes. The anodic oxidation of TAA showed well-defined limiting currents or current peaks and a good linearity of the amperometric signal vs. concentration plots. The analytical parameters of sensitivity, RSD and LOD, obtained under various experimental conditions, suggest the suitability of the BDD electrode for electroanalytical purposes. Low fouling effects, good reproducibility and stability, as well as the sharpness of the amperometric signals in both unbuffered/ buffered acidic or neutral media, highly superior to those obtained using a glassy carbon (GC electrode, recommend the unmodified BDD electrode as a promising potential amperometric sensor for environmental applications, regarding the direct anodic determination of TAA in aqueous media.

  1. Increasing amperometric biosensor sensitivity by length fractionated single-walled carbon nanotubes

    DEFF Research Database (Denmark)

    Tasca, Federico; Gorton, Lo; Wagner, Jakob Birkedal

    2008-01-01

    In this work the sensitivity-increasing effect of single-walled carbon nanotubes (SWCNTs) in amperometric biosensors, depending on their average length distribution, was studied. For this purpose the SWCNTs were oxidatively shortened and subsequently length separated by size exclusion chromatogra......In this work the sensitivity-increasing effect of single-walled carbon nanotubes (SWCNTs) in amperometric biosensors, depending on their average length distribution, was studied. For this purpose the SWCNTs were oxidatively shortened and subsequently length separated by size exclusion...... chromatography. Transmission electron micrographs of different fractions of SWCNTs were collected. Diaphorase ``wired'' to an osmium redox polymer was blended with the shortened SWCNTs of different lengths. Depending on the average length of the SWCNTs the sensitivity of the amperometric biosensor model system...... limit was 1 mu M. The biosensor exhibited excellent electrocatalytic properties. Even at relatively high NADH concentrations the oxidative current was limited by the diffusion rate of NADH. (C) 2008 Elsevier B.V. All rights reserved....

  2. 基于聚邻苯二胺/石墨烯修饰电极的肠道病毒71型(EV71)电化学免疫传感器%Electrochemical Immunosensor for Detection of Human Enterovirus 71 Based on Graphene-Poly (o-Phenylenediamine) Modified Electrode

    Institute of Scientific and Technical Information of China (English)

    谢爱娟; 周民; 罗士平; 尹俊; 孔泳

    2013-01-01

    采用改性的Hummers法制备石墨烯氧化物(GO),并将其滴涂在玻碳电极(GCE)表面.通过电化学还原将GO还原为石墨烯,并进一步采用电化学聚合法在石墨烯表面形成聚邻苯二胺(PoPD)膜,从而制备了PoPD/石墨烯修饰GCE.将EV71抗体固定在修饰电极表面,制备新型的电化学免疫传感器.当发生免疫反应时,由于EV71抗原与抗体生成的免疫复合物阻碍了电子的传递,PoPD的氧化峰电流下降.当抗原的浓度在0.1-80ng/mL范围内,PoPD氧化峰电流的降低值与抗原的浓度成正比.该免疫传感器对EV71的检测限为0.08ng/mL(信噪比为3).%Graphene oxide (GO) was prepared by a modified Hummers method and dropped onto the surface of a glassy carbon electrode (GCE). The GO was electrochemically reduced to graphene, and poly (o -phenylenediamine) (PoPD) film was then electrosynthesized on the surface of graphene to prepare the PoPD/graphene modified GCE. A novel electrochemical immunosensor was obtained by immobilizing anti - EV71 onto the surface of this modified electrode. When the immunoreaction between anti - EV71 and EV71 antigen in the solution took place, the electron transfer would be hindered by the produced immuno-complex and thus the oxidation peak current of PoPD would decrease. The decrease in the oxidation peak current of PoPD was proportional to the concentration of EV71 antigen in the range from 0. 1 to 80ng/mL, and the detection limit was as low as 0. 08ng/mL (signal-to-noise ratio of 3).

  3. Microfabricated biosensor for the simultaneous amperometric and luminescence detection and monitoring of Ochratoxin A.

    Science.gov (United States)

    Tria, Scherrine A; Lopez-Ferber, David; Gonzalez, Catherine; Bazin, Ingrid; Guiseppi-Elie, Anthony

    2016-05-15

    The low molecular weight hapten, Ochratoxin A (OTA), is a natural carcinogenic mycotoxin produced by Aspergillus and Penicillium fungi and so it commonly appears in wines, other foods, and in the environment. An amperometric biosensor has been developed that uses the immobilized synthetic peptide, NFO4; which possesses a high binding affinity and thus provides for molecular recognition of OTA; simulating the mycotoxin-specific antibody. Biotransducers were produced from a microlithographically fabricated electrochemical cell-on-a-chip that uses the microdisc electrode array working electrode format augmented with microporous graphitized carbon (MGC) that was electrodeposited within a poly(aniline-co-meta-aminoaniline) electroconductive polymer layer. A redox mediator, iron-nickel hexacyanoferrate (Fe|NiHCF) was amperometrically deposited onto the MGC. The device was then dip-coated with monomer cocktail that yielded poly(HEMA-co-AEMA) foam that was prepared in-situ by UV crosslinking and by sequentially freezing followed by freeze drying of the chip to yield a 3-D support for the chelation of Zn(2+) ions (ZnCl2) and the subsequent immobilization of N-terminus his-tagged peptide, NFO4. To conduct the biosensors assay, HRP conjugated OTA was added to the free OTA solutions and together competitively incubated on the biospecific MDEA ECC 5037-Pt|MGC|HCF|Hydrogel-NFO4 biotransducer. The amperometric response to peroxide was determined after 5 min of enzymatic reaction following addition of standard substrate H2O2/luminol. Simultaneous analysis of light emission signals (λmax=425 nm) allowed direct comparison of amperometric and luminescence performance. Using chitosan foam and a luminescence bioassay we obtained maximum inhibition at 10 μg L(-1) and half inhibition occurred at 2.1 μg L(-1). Using poly(HEMA-co-AEMA) hydrogel and an amperometric bioassay (50s) we obtained maximum inhibition at 10 μg L(-1) and half inhibition occurred at 2.8 μg L(-1).

  4. Dual-responsive immunosensor that combines colorimetric recognition and electrochemical response for ultrasensitive detection of cancer biomarkers.

    Science.gov (United States)

    Hong, Wooyoung; Lee, Sooyeon; Cho, Youngnam

    2016-12-15

    We developed a nanoroughened, biotin-doped polypyrrole immunosensor for the detection of tumor markers through dual-signal (electrochemical and colorimetric) channels, electrochemical and colorimetric, that demonstrates remarkable analytical performance. A rapid, one-step electric field-mediated method was employed to fabricate the immunosensor with nanoscale roughness by simply modulating the applied electrical potential. We demonstrated the successful detection of three tumor markers (CA125, CEA, and PSA) via the double enzymatic signal amplifications in the presence of a target antigen, ultimately leading to desired diagnostic accuracy and reliability. The addition of multiple horseradish peroxidase (HRP)- and antibody-labeled nanoparticles greatly amplified the signal and simplified the measurement of cancer biomarker proteins by sequentially magnifying electrochemical and colorimetric signals in a single platform. The two parallel assays performed using the proposed immunosensor have yielded highly consistent and reproducible results. Additionally, for the analysis of plasma samples in a clinical setting, the values obtained with our immunosensor were validated by correlating the results with those of a standard radioimmunoassay (RIA), which obtained very similar clinically valid responses.

  5. Application of a Label-Free Immunosensor for White Spot Syndrome Virus (WSSV) in Shrimp Cultivation Water.

    Science.gov (United States)

    Waiyapoka, Thanyaporn; Deachamag, Panchalika; Chotigeat, Wilaiwan; Bunsanong, Nittaya; Kanatharana, Proespichaya; Thavarungkul, Panote; Loyprasert-Thananimit, Suchera

    2015-10-01

    White spot syndrome virus (WSSV) is a major pathogen affecting the shrimp industry worldwide. In a preliminary study, WSSV binding protein (WBP) was specifically bound to the VP26 protein of WSSV. Therefore, we have developed the label-free affinity immunosensor using the WBP together with anti-GST-VP26 for quantitative detection of WSSV in shrimp pond water. When the biological molecules were immobilized on a gold electrode to form a self-assembled monolayer, it was then used to detect WSSV using a flow injection system with optimized conditions. Binding between the different copies of WSSV and the immobilized biological molecules was detected by an impedance change (ΔZ″) in real time. The sensitivity of the developed immunosensor was in the linear range of 1.6 × 10(1)-1.6 × 10(6) copies/μl. The system was highly sensitive for the analysis of WSSV as shown by the lack of impedance change when using yellow head virus (YHV). The developed immunosensor could be reused up to 37 times (relative standard deviation (RSD), 3.24 %) with a good reproducibility of residual activity (80-110 %). The immunosensor was simple to operate, reliable, reproducible, and could be applied for the detection and quantification of WSSV in water during shrimp cultivation.

  6. Catalytic activity of iron hexacyanoosmate(II) towards hydrogen peroxide and nicotinamide adenine dinucleotide and its use in amperometric biosensors

    Energy Technology Data Exchange (ETDEWEB)

    Kotzian, Petr; Janku, Tereza [Department of Analytical Chemistry, University of Pardubice, Nam. Cs. Legii 565, CZ-532 10 Pardubice (Czech Republic); Kalcher, Kurt [Institute of Chemistry - Analytical Chemistry, Karl-Franzens University, Universitaetsplatz 1, A-8010 Graz (Austria); Vytras, Karel [Department of Analytical Chemistry, University of Pardubice, Nam. Cs. Legii 565, CZ-532 10 Pardubice (Czech Republic)], E-mail: karel.vytras@upce.cz

    2007-09-19

    Hydrogen peroxide and nicotinamide adenine dinucleotide (NADH) may be determined amperometrically using screen-printed electrodes chemically modified with iron(III) hexacyanoosmate(II) (Osmium purple) in flow injection analysis (FIA). The determination is based on the exploitation of catalytic currents resulting from the oxidation/reduction of the modifier. The performance of the sensor was characterized and optimized by controlling several operational parameters (applied potential, pH and flow rate of the phosphate buffer). Comparison has been made with analogous complexes of ruthenium (Ruthenium purple) and iron (Prussian blue). Taking into account the sensitivity and stability of corresponding sensors, the best results were obtained with the use of Osmium purple. The sensor exhibited a linear increase of the amperometric signal with the concentration of hydrogen peroxide in the range of 0.1-100 mg L{sup -1} with a detection limit (evaluated as 3{sigma}) of 0.024 mg L{sup -1} with a R.S.D. 1.5% for 10 mg L{sup -1} H{sub 2}O{sub 2} under optimized flow rate of 0.4 mL min{sup -1} in 0.1 M phosphate buffer carrier (pH 6) and a working potential of +0.15 V versus Ag/AgCl. Afterwards, a biological recognition element - either glucose oxidase or ethanol dehydrogenase - was incorporated to achieve a sensor facilitating the determination of glucose or ethanol, respectively. The glucose sensor gave linearity between current and concentration in the range from 1 to 250 mg L{sup -1} with a R.S.D. 2.4% for 100 mg L{sup -1} glucose, detection limit 0.02 mg L{sup -1} (3{sigma}) and retained its original activity after 3 weeks when stored at 6 deg. C. Optimal parameters in the determination of ethanol were selected as: applied potential +0.45 V versus Ag/AgCl, flow rate 0.2 mL min{sup -1} in 0.1 M phosphate buffer carrier (pH 7). Different structural designs of the ethanol sensor were tested and linearity obtained was up to 1000 mg L{sup -1} with a maximum R.S.D. of 5

  7. Comparison between amperometric and true potentiometric end-point detection in the determination of water by the Karl Fischer method.

    Science.gov (United States)

    Cedergren, A

    1974-06-01

    A rapid and sensitive method using true potentiometric end-point detection has been developed and compared with the conventional amperometric method for Karl Fischer determination of water. The effect of the sulphur dioxide concentration on the shape of the titration curve is shown. By using kinetic data it was possible to calculate the course of titrations and make comparisons with those found experimentally. The results prove that the main reaction is the slow step, both in the amperometric and the potentiometric method. Results obtained in the standardization of the Karl Fischer reagent showed that the potentiometric method, including titration to a preselected potential, gave a standard deviation of 0.001(1) mg of water per ml, the amperometric method using extrapolation 0.002(4) mg of water per ml and the amperometric titration to a pre-selected diffusion current 0.004(7) mg of water per ml. Theories and results dealing with dilution effects are presented. The time of analysis was 1-1.5 min for the potentiometric and 4-5 min for the amperometric method using extrapolation.

  8. Nanostructuring of hierarchical 3D cystine flowers for high-performance electrochemical immunosensor.

    Science.gov (United States)

    Pandey, Chandra Mouli; Sumana, Gajjala; Tiwari, Ida

    2014-11-15

    Here, we report a simple and reproducible method for large scale fabrication of novel flower and palm-leaf like 3D cystine microstructures (CMs) with high uniformity having a size of 50 µm and 10 µm respectively, through a facile aqueous solution route as a function of pH and concentration. In a proof-of-concept study, the 3D CMs have been further explored to fabricate a label-free high-performance electrochemical immunosensor by immobilizing monoclonal antibodies. Electrochemical methods were employed to study the stepwise modification of the system and the electronic transduction for the detection. The fabricated immunosensor design demonstrates high performance with enhanced sensitivity (4.70 cfu ml(-1)) and linear sensing range from 10 to 3 x 10(9) cfu ml(-1) a long shelf-life (35 days) and high selectivity over other bacterial pathogens. The enhanced performance originates from a novel nanostructuring in which the CMs provide higher surface coverage for the immobilization of antibodies providing excellent electronic/ionic conductivity which result in the enhanced sensitivity.

  9. A sensitive electrochemical impedance immunosensor for determination of malachite green and leucomalachite green in the aqueous environment.

    Science.gov (United States)

    Zhu, Dan; Li, Qiangqiang; Pang, Xiumei; Liu, Yue; Wang, Xue; Chen, Gang

    2016-08-01

    Application of malachite green (MG) and leucomalachite green (LMG) in fish farm water causes an environmental problem. This study proposes for the first time a sensitive and convenient electrochemical impedance spectroscopy (EIS) method for determining MG and LMG by a bovine serum albumin-decorated gold nanocluster (BSA-AuNC)/antibody composite film-based immunosensor. In order to improve the analytical performance, the glassy carbon electrode (GCE) was modified with 1, 4-phenylenediamine to form a stable layer, and then, BSA-AuNCs were covalently bound to the GCE. An adequate quantity of the polyclonal antibody of LMG was immobilized onto the surface of the BSA-AuNCs by the chemical reaction of EDC/NHS. The sensors can respond to the specific target based on specific covalent bonding. The experimental parameters, such as the pH, incubating concentration, and time, have been investigated and optimized. The calibration curve for LMG was linear in the range of 0.1~10.0 ng/mL with the limit of detection (LOD) 0.03 ng/mL. Furthermore, the sum of MG and LMG was detected in fish farm water by MG reduction. The recovery was between 89.7 % and 99.2 % in spiked samples. The EC sensor method was also compared with the ELISA method and validated by the LC-MS/MS method, which proves its great promise as a field instrument for the rapid monitoring of MG and LMG pollution. Graphical abstract 1, 4-Phenylenediamine and BSA-AuNC/antibody-decorated glassy carbon electrodes have been used for the impedimetric detection of the sum of malachite green and leucomalachite green via specific immuno-binding.

  10. Analysis of polyphenols in white wine by CZE with amperometric detection using carbon nanotube-modified electrodes.

    Science.gov (United States)

    Moreno, Mónica; Arribas, Alberto Sánchez; Bermejo, Esperanza; Zapardiel, Antonio; Chicharro, Manuel

    2011-04-01

    A method for the simultaneous detection of five polyphenols (caffeic, chlorogenic, ferulic and gallic acids and (+)-catechin) by CZE with electrochemical detection was developed. Separation of these polyphenols was performed in a 100 mM borate buffer (pH 9.2) within 15 min. Under optimized separation conditions, the performance of glassy carbon (GC) electrodes modified with multiwalled carbon nanotube layer obtained from different dispersions was examined. GC electrode modified with a dispersion of multi-walled carbon nanotubes (CNT) in polyethylenimine has proven to be the most suitable CNT-based electrode for its application as amperometric detector for the CZE separation of the studied compounds. The excellent electrochemical properties of this electrode allowed the detection of the selected polyphenols at +200 mV and improved the efficiency and the resolution of their CZE separation. Limits of detection below 3.1 μM were obtained with linear ranges covering the 10⁻⁵ to 10⁻⁴  M range. The proposed method has been successfully applied for the detection (ferulic, caffeic and gallic acids and (+)-catechin) and the quantification (gallic acid and (+)-catechin) of polyphenols in two different white wines without any preconcentration step. A remarkable signal stability was observed on the electrode performance despite the presence of potential fouling substances in wine.

  11. Classification of Spanish white wines using their electrophoretic profiles obtained by capillary zone electrophoresis with amperometric detection.

    Science.gov (United States)

    Arribas, Alberto Sánchez; Martínez-Fernández, Marta; Moreno, Mónica; Bermejo, Esperanza; Zapardiel, Antonio; Chicharro, Manuel

    2014-06-01

    A method was developed for the simultaneous detection of eight polyphenols (t-resveratrol, (+)-catechin, quercetin and p-coumaric, caffeic, sinapic, ferulic, and gallic acids) by CZE with electrochemical detection. Separation of these polyphenols was achieved within 25 min using a 200 mM borate buffer (pH 9.4) containing 10% methanol as separation electrolyte. Amperometric detection of polyphenols was carried out with a glassy carbon electrode (GCE) modified with a multiwalled carbon nanotubes (CNT) layer obtained from a dispersion of CNT in polyethylenimine. The excellent electrochemical properties of this modified electrode allowed the detection and quantification of the selected polyphenols in white wines without any pretreatment step, showing remarkable signal stability despite the presence of potential fouling substances in wine. The electrophoretic profiles of white wines, obtained using this methodology, have proven to be useful for the classification of these wines by means of chemometric multivariate techniques. Principal component analysis and discriminant analysis allowed accurate classification of wine samples on the basis of their grape varietal (verdejo and airén) using the information contained in selected zones of the electropherogram. The utility of the proposed CZE methodology based on the electrochemical response of CNT-modified electrodes appears to be promising in the field of wine industry and it is expected to be successfully extended to classification of a wider range of wines made of other grape varietals.

  12. Poly(3,4-ethylenedioxythiophene)-based glucose biosensors

    NARCIS (Netherlands)

    Kros, A.; Hövell, W.F.M. van; Sommerdijk, N.A.J.M.; Nolte, R.J.M.

    2001-01-01

    Amperometric biosensors for the recognition of glucose oxidase (GOx) based on poly(3,4-ethylenedioxythiophene) (PEDOT) were fabricated for the first time. The resulting biosensor has potential applications for long-term glucose measurements.

  13. Separation and determination of alditols and sugars by high-pH anion-exchange chromatography with pulsed amperometric detection

    DEFF Research Database (Denmark)

    Andersen, Rikke; Sørensen, A.

    2000-01-01

    Carbohydrates such as alditols (polyols or sugar alcohols), monosaccharides and disaccharides are separated as anions by anion-exchange chromatography with a sodium hydroxide eluent, MA1 CarboPac column and pulsed amperometric detection. We report a high-pH anion-exchange chromatographic......-pulsed amperometric detection (HPAEC-PAD) method that determines all the polyols used as food additives in food products and the most commonly found mono- and disaccharides on a routine basis. The linearity, repeatability, internal reproducibility and accuracy are described. The applicability of the method has been...

  14. Creation of a Databank for Content of Antioxidants in Food Products by an Amperometric Method

    Directory of Open Access Journals (Sweden)

    Polina A. Fedina

    2010-10-01

    Full Text Available Oxidative stress, i.e. excessive content of reactionary, oxygen, and nitrogen compounds (ROAC, including free radicals, is one of the causes of various dangerous diseases as well as premature aging. The adverse effect of free radicals can be neutralized by antioxidants. In order to carry out antioxidant therapy, one needs to know the contents of antioxidants in food products. We have created the databank for the contents of antioxidants in 1,140 food products, beverages, etc. Apart from water-soluble antioxidants, fat-soluble antioxidants in dairy and fish products, cacao, chocolate, nuts etc. were determined for the first time using an amperometric method.

  15. Horseradish peroxidase-catalyzed polymerization of L-DOPA for mono-/bi-enzyme immobilization and amperometric biosensing of H2O2 and uric acid.

    Science.gov (United States)

    Dai, Mengzhen; Huang, Ting; Chao, Long; Xie, Qingji; Tan, Yueming; Chen, Chao; Meng, Wenhua

    2016-01-01

    Horseradish peroxidase (HRP)-catalyzed polymerization of L-DOPA (vs. dopamine) in the presence of H2O2 (and uricase (UOx)) was exploited to immobilize mono-/bi-enzymes for hydroquinone-mediated amperometric biosensing of H2O2 and uric acid (UA). The relevant polymeric biocomposites (PBCs) were prepared in phosphate buffer solution containing HRP and L-DOPA (or plus UOx) after adding H2O2. The mono-/bi-enzyme amperometric biosensors were prepared simply by casting some of the PBCs on Au-plated Au (Au(plate)/Au) electrodes, followed by coating with an outer-layer chitosan (CS) film for each. UV-vis spectrophotometry, scanning electron microscopy, cyclic voltammetry and electrochemical impedance spectroscopy were used for film characterization and/or process monitoring. The HRP immobilized by enzyme catalysis well preserved its bioactivity, as confirmed by UV-vis spectrophotometry. Under optimized conditions, the monoenzyme CS/HRP-poly(L-DOPA) (PD)/Au(plate)/Au electrode potentiostated at -0.1V responded linearly to H2O2 concentration from 0.001 to 1.25mM with a sensitivity of 700μA mM(-1)cm(-2) and a limit of detection (LOD) of 0.1μM, and the bienzyme CS/UOx-HRP-PD/Au(plate)/Au electrode at -0.1V responded linearly to UA concentration from 0.001 to 0.4mM with a sensitivity of 349μA mM(-1)cm(-2) and a LOD of 0.1μM. The mono-/bi-enzyme biosensors based on biosynthesized PD performed better than many reported analogues and those based on similarly biosynthesized polydopamine.

  16. Mycobacterium tuberculosis 38 kDa Antigen Purification and Potential Diagnostic Use by Piezoelectric Immunosensors

    Directory of Open Access Journals (Sweden)

    Paula A. MARÍN

    2014-10-01

    Full Text Available The selection and procurance of the immunoreagents used in the immunoassay and biofunctionalisation transducer strategy, are a key in the piezoelectric immunosensor development for the detection of tuberculosis (TB. Many have reported the use of 38kDa protein antigen (Ag38kDa from Mycobacterium tuberculosis (Mtb such as good biomarker of TB disease and compliance with physical and biochemical characteristics to be immobilized by self-assembled monolayers (SAMs, in the gold electrode of piezoelectrics crystals surfaces. A piezoelectric immunosensor developed from purified native antigens of Mtb may be an alternative simple method for detection of Mtb with speed and reusable advantages, contributing to the control and early treatment of disease. In this paper, the purification process of Ag38kDa Mtb from secretory proteins filtered culture (CFP from Mtb is presented as an immunoreactive with potential application in the detection of Mtb by piezoelectric immunosensors. Functionalized crystals were obtained by using the modified self-assembled monolayers (SAMs technique, with purified native antigen and CFP. The functionalized surfaces were qualitatively characterized using atomic force microscopy (AFM in order to validate the immobilization protocol optimal conditions for secretion antigens from Mtb. These modified crystals may be coupled to piezoelectric immunosensor characterization system for detecting of Mtb by a direct competition immunoassay. PURIFICACIÓN DEL ANTÍGENO 38kDa DE Mycobacterium tuberculosis Y SU POTENCIAL USO EN DIAGNÓSTICO MEDIANTE INMUNOSENSORES PIEZOELÉCTRICOSUn paso crucial en el desarrollo de un inmunosensor piezoeléctrico para la detección de tuberculosis (TB, es la selección y obtención de los inmunoreactivos empleados en el inmunoensayo y la estrategia para la biofuncionalización del transductor. Diversos estudios han reportado el uso del antígeno proteico 38kDa (Ag38kDa de Mycobacterium

  17. Development of 170 MHz Electrodeless Quartz-Crystal Microbalance Immunosensor with Nonspecifically Immobilized Receptor Proteins

    Science.gov (United States)

    Ogi, Hirotsugu; Nagai, Hironao; Fukunishi, Yuji; Yanagida, Taiji; Hirao, Masahiko; Nishiyama, Masayoshi

    2010-07-01

    Staphylococcus aureus protein A (SPA) shows high nonspecific binding affinity on a naked quartz surface, and it can be used as the receptor protein for detecting immunoglobulin G (IgG), the most important immunoglobulin. The immunosensor ability, however, significantly depends on the immobilization procedure. In this work, the effect of the nonspecific immobilization procedure on the sensor sensitivity is studied using a home-built electrodeless quartz-crystal microbalance (QCM) biosensor. The pure-shear vibration of a 9.7-µm-thick AT-cut quartz plate is excited and detected in liquids by the line antenna located outside the flow channel. SPA molecules are immobilized on the quartz surfaces, and human IgG is injected to monitor the binding reaction between SPA and IgG. This study reveals that a long (nearly 24 h) immersion procedure is required for immobilizing SPA to achieve the tight biding with the quartz surfaces.

  18. Screening and confirmatory methods for the analysis of macrocyclic lactone mycotoxins by CE with amperometric detection.

    Science.gov (United States)

    Arribas, Alberto Sánchez; Bermejo, Esperanza; Zapardiel, Antonio; Téllez, Helena; Rodríguez-Flores, Juana; Zougagh, Mohammed; Ríos, Angel; Chicharro, Manuel

    2009-02-01

    A simple analytical scheme for the screening and quantification of zearalenone and its metabolites, alpha-zearalenol and beta-zearalenol, is reported. Extracts from maize flour samples were collected by supercritical fluid extraction and afterwards, they were analyzed by CE with amperometric detection. This scheme allowed a rapid and reliable identification of contaminated flour samples according to the reference value established for zearalenone by directive 2005/38/EC (200 microg/kg). The sample screening method was carried out by CZE using 25 mM borate separation buffer at pH 9.2 and 25.0 kV as separation voltage, monitoring the amperometric signal at +700 mV with a carbon paste electrode. In this way, total amount of mycotoxins was determined and samples were processed in 4 min with a detection limit of 12 microg/L, enough to discriminate between positive (more than 200 microg/L total mycotoxins) and negative samples (less than 200 microg/L total mycotoxins). Positive samples were then subjected to CZE separation and quantification of each analyte was done with 50 mM borate running buffer modified with 30% methanol at pH 9.7 and 17.5 kV as separation voltage. Under these conditions, separation was achieved in 15 min with detection limits from 20 to 35 microg/L for each analyte.

  19. Comparison of Electrochemical Immunosensors and Aptasensors for Detection of Small Organic Molecules in Environment, Food Safety, Clinical and Public Security

    Directory of Open Access Journals (Sweden)

    Benoit Piro

    2016-02-01

    Full Text Available We review here the most frequently reported targets among the electrochemical immunosensors and aptasensors: antibiotics, bisphenol A, cocaine, ochratoxin A and estradiol. In each case, the immobilization procedures are described as well as the transduction schemes and the limits of detection. It is shown that limits of detections are generally two to three orders of magnitude lower for immunosensors than for aptasensors, due to the highest affinities of antibodies. No significant progresses have been made to improve these affinities, but transduction schemes were improved instead, which lead to a regular improvement of the limit of detections corresponding to ca. five orders of magnitude over these last 10 years. These progresses depend on the target, however.

  20. Determination of Roxithromycin Tablets by Capillary Electrophoresis Employing Non-aqueous Media with Square-wave Amperometric Detection

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    A new method of determination for roxithromycin tablets by non-aqueous capillary electrophoresis (NACE) with square-wave amperometric detection was carried out. Several parameters affecting the NACE-AD determination were studied. The data was modified by spline wavelet least square (SWLS). The method is simple, rapid and highly reliable for routine analysis.

  1. Stainless steel modified with an aminosilane layer and gold nanoparticles as a novel disposable substrate for impedimetric immunosensors.

    Science.gov (United States)

    Rezaei, Behzad; Havakeshian, Elaheh; Ensafi, Ali A

    2013-10-15

    In this work, stainless steel (SS) was used as a substrate to fabricate an inexpensive and disposable impedimetric immunosensor. SS surface was modified with a stable thin layer of 3-aminopropyltriethoxysilane (APTES), and followed by electrodeposition of gold nanoparticles (GNPs). The morphology and size of the electrodeposited GNPs were studied using scanning electron microscopy (SEM) and atomic force microscopy (AFM). The interfacial properties of the SS electrode after each modification step were characterized by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) in a solution containing [Fe(CN)₆]³⁻/⁴⁻ as a redox probe. The results indicated that APTES layer was successfully formed on the electrode surface and GNPs enhanced the conductivity and sensitivity of the electrode. The applicability of the proposed assembled electrode in electrochemical immunosensors was followed by immobilizing doxorubicin-specific monoclonal antibodies onto the GNP-modified electrode to determine doxorubicin concentration using the EIS technique. The relative charge transfer resistance was found to increase linearly with doxorubicin concentration in two ranges from 2.5 to 30.0 and 30.0 to 100.0 pg mL⁻¹. The detection limit of the immunosensor was 1.7 pg mL⁻¹ (3s(b)/m) doxorubicin. The satisfactory results were obtained from determination of doxorubicin concentrations in spiked human serum samples. The recoveries were in the range of 88.0-122.2%. These results indicate that modified SS electrodes are promising sensing elements to construct economical electrochemical immunosensors for routine quantitative analyses.

  2. Label-Free Detection of Chondroitin Sulphate Proteoglycan 4 by a Polyaniline/Graphene Nanocomposite Functionalized Impedimetric Immunosensor

    Directory of Open Access Journals (Sweden)

    JingJing Fu

    2016-01-01

    Full Text Available The chondroitin sulphate proteoglycan 4 (CSPG4, also known as high molecular weight-melanoma associated antigen (HMW-MAA, is a tumor-associated antigen that is expressed in more than 85% of surgically removed melanoma lesions but has restricted distribution in normal tissues. The diagnostic and therapeutic value of CSPG4 drives a need for sensitive and low-cost detection approaches. To this end, we developed a polyaniline/graphene oxide nanocomposite (PANI@GO that was electrochemically codeposited on indium tin oxide (ITO electrode. Glutaraldehyde mediated the covalent immobilization of CSPG4 specific antibody mAbD2.8.5 to construct a CSPG4 immunosensor using cell culture media and cell lysate as samples. The fully assembled impedimetric immunosensor was used to detect CSPG4 in CSPG4-positive cell lines M14/CSPG4 and MV3. No impedance signal changes could be observed from CSPG4-negative cell lines M14 and mAbMk2-23 showing the specificity of the CSPG4-impedimetric immunosensor. This low-cost, simple, and label-free analytical method is an alternative to enzyme-linked immunosorbent assay and flow cytometry in screening of CSPG4 in complex biological samples.

  3. Surface-enhanced fluorescence immunosensor using Au nano-crosses for the detection of microcystin-LR.

    Science.gov (United States)

    Li, Yun; Sun, Jiadi; Wu, Longyun; Ji, Jian; Sun, Xiulan; Qian, Yongzhong

    2014-12-15

    A surface-enhanced fluorescence (SEF) immunosensor for the detection of microcystin-LR was developed using Au nano-crosses as fluorescence enhancement nanoparticles and cy5 as a fluorescence label molecule. The SEF effects of cy5 in the proximity of Au nanorods and gold nano-crosses was investigated by using Au nanorods or nano-crosses coated negative-charged glass surfaces. Fluorescence measurements indicated that SEF was influenced by the size, shape and distribution of the Au nanoparticles, with an appropriate spacer layer between the Au nanoparticles and the cy5. The enhancement factor was from 2.3- to 35-fold. Under optimal conditions, the SEF immunosensor exhibited a good linear response at microcystin-LR concentrations of 0.02-16 ng mL(-1) (R(2)=0.9981). The limit of detection was 0.007 ng mL(-1) with little adsorption of microcystin-RR, microcystin-LW, and microcystin-LF. High microcystin-LR recoveries were obtained from naturally contaminated fish samples. The SEF immunosensor allows the reliable detection of microcystin-LR in seafood, and has potential in simple, sensitive detection applications.

  4. Mediatorless amperometric glucose biosensing using 3-aminopropyltriethoxysilane-functionalized graphene.

    Science.gov (United States)

    Zheng, Dan; Vashist, Sandeep Kumar; Al-Rubeaan, Khalid; Luong, John H T; Sheu, Fwu-Shan

    2012-09-15

    A mediatorless glucose biosensor was developed by the immobilization of glucose oxidase (GOx) to graphene-functionalized glassy carbon electrode (GCE). The surface of GCE was functionalized with graphene by incubating it with graphene dispersed in 3-aminopropyltriethoxysilane (APTES), which acted both as a dispersion agent for graphene and as an amine surface modification agent for GCE and graphene. This was followed by the covalent binding of GOx to graphene-functionalized GCE using 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC) based crosslinking. Graphene provided signal enhancement by providing greater surface area for GOx binding, while APTES-functionalization led to a higher GOx immobilization density by providing free amino groups for crosslinking. The developed biosensor used a redox potential of -0.45 V (vs. Ag/AgCl) for detecting glucose in the diabetic pathophysiological range 0.5-32 mM. There was no interference from endogenous electroactive substances and drug metabolites. The developed biosensor was further validated for detecting blood glucose in commercial artificial blood glucose linearity standards in the range 1.4-27.9 mM. Therefore, it is ideal for diabetic blood glucose monitoring. The developed bioanalytical procedure for preparation of GOx-bound graphene-functionalized GCEs had high production reproducibility and high storage stability, which is appropriate for the commercial mass production of enzyme-bound electrodes.

  5. Determination of neomycin sulfate and impurities using high-performance anion-exchange chromatography with integrated pulsed amperometric detection.

    Science.gov (United States)

    Hanko, Valoran P; Rohrer, Jeffrey S

    2007-01-01

    Neomycin B is one of a class of aminoglycoside antibiotics that lack a good chromophore, and is therefore difficult to determine using reversed-phase HPLC with absorbance detection. This is especially true for determining the quantity of each impurity. We show that neomycin sulfate and its major impurities, including neamine (neomycin A), can be separated on a strong anion-exchange column using a weak potassium hydroxide eluent (2.40 mM) at a column temperature of 30 degrees C, and directly detected by integrated pulsed amperometric detection (IPAD). The resolution (United States Pharmacopeia (USP) definition) between neomycin B and the closest major impurity ranged from 6.56 and 7.45 over 10 days of consecutive analysis (7.24+/-0.10, n=836 injections). Due to the difficulty of producing weak hydroxide eluents of the required purity (i.e. carbonate-free), this method depends on automatic eluent generation to ensure method ruggedness. This method exhibited good long-term (10 days, 822 injections) retention time stability with a R.S.D. of 0.6%. Peak area R.S.D. (10 microM) was 1.3%. Method robustness was evaluated by intentionally varying the flow rate, eluent concentration, column temperature, and column. The spike recoveries of neomycin B from extractions of three different topical ointments and cream formulations ranged from 95 to 100%. The measured concentration of neomycin B in these formulations ranged from 119 to 154% of the label concentration. The R.S.D. for the measured concentration of one of the formulations tested over three separate days, n=11 extracts, was 3.2%. Based on the results of these evaluations, we believe this method can be used for neomycin sulfate identity, assay, and purity.

  6. Pristine multi-walled carbon nanotubes/SDS modified carbon paste electrode as an amperometric sensor for epinephrine.

    Science.gov (United States)

    Thomas, Tony; Mascarenhas, Ronald J; D' Souza, Ozma J; Detriche, Simon; Mekhalif, Zineb; Martis, Praveen

    2014-07-01

    An amperometric sensor for the determination of epinephrine (EP) was fabricated by modifying the carbon paste electrode (CPE) with pristine multi-walled carbon nanotubes (pMWCNTs) using bulk modification followed by drop casting of sodium dodecyl sulfate (SDS) onto the surface for its optimal potential application. The modified electrode showed an excellent electrocatalytic activity towards EP by decreasing the overpotential and greatly enhancing the current sensitivity. FE-SEM images confirmed the dispersion of pMWCNTs in the CPE matrix. EDX analysis ensured the surface coverage of SDS. A comparative study of pMWCNTs with those of oxidized MWCNTs (MWCNTsOX) modified electrodes reveals that the former is the best base material for the construction of the sensor with advantages of lower oxidation overpotential and the least background current. The performance of the modified electrode was impressive in terms of the least charge transfer resistance (Rct), highest values for diffusion coefficient (DEP) and standard heterogeneous electron transfer rate constant (k°). Analytical characterization of the modified electrode exhibited two linear dynamic ranges from 1.0×10(-7) to 1.0×10(-6)M and 1.0×10(-6) to 1.0×10(-4)M with a detection limit of (4.5±0.18)×10(-8)M. A 100-fold excess of serotonin, acetaminophen, folic acid, uric acid, tryptophan, tyrosine and cysteine, 10-fold excess of ascorbic acid and twofold excess of dopamine do not interfere in the quantification of EP at this electrode. The analytical applications of the modified electrode were demonstrated by determining EP in spiked blood serum and adrenaline tartrate injection. The modified electrode involves a simple fabrication procedure, minimum usage of the modifier, quick response, excellent stability, reproducibility and anti-fouling effects.

  7. Dual enzymatic biosensor for simultaneous amperometric determination of histamine and putrescine.

    Science.gov (United States)

    Henao-Escobar, W; del Torno-de Román, L; Domínguez-Renedo, O; Alonso-Lomillo, M A; Arcos-Martínez, M J

    2016-01-01

    A disposable electrodic system consisting of two working electrodes connected in array mode has been developed for the simultaneous determination of histamine (His) and putrescine (Put). Histamine deshydrogenase and putrescine oxidase enzymes were respectively immobilized by crosslinking on each working screen-printed electrode, both modified with tetrathiafulvalene. The dual system allowed the simultaneous amperometric determination of both species by measuring the oxidation current of the mediator in each working electrode. The effect of other potentially interfering biogenic amines was also evaluated. The capability of detection was of 8.1 ± 0.7 for His and 10 ± 0.6 μM for Put. The precision in terms of relative standard deviation was of 3.5% and 6.7% for His and Put, respectively. The developed biosensor was successfully applied to the determination of His and Put in different food samples.

  8. Determination of neomycin in water samples by high performance anion chromatography with pulsed amperometric detection

    Institute of Scientific and Technical Information of China (English)

    Bin Guan; Dong Xing Yuan

    2007-01-01

    A simple, fast and reliable method, using high performance anion chromatography with pulsed amperometric detection, had been developed for the analysis of neomycin in water samples. The elution and separation were carried out with an isocratic mobile phase, containing 10 mmol/L NaOH. The influence of the concentration and pH of the mobile phase on the separation and detection was investigated. A quadruple-potential waveform used for the detection was optimized. The detection limit of neomycin was down to 0.027 μg/mL. The linearity of neomycin calibration curve ranged from 0.050 to 0.505 μg/mL with correlation coefficient of0.9997. R.S.D. (n= 11) was 4.0%.

  9. Rapid measurement of free cyanide in liquor by ion chromatography with pulsed amperometric detection.

    Science.gov (United States)

    Wu, Wenlin; Xiao, Quanwei; Zhang, Ping; Ye, Mei; Wan, Yuping; Liang, Hengxing

    2015-04-01

    This study investigated the measurement of free cyanide in liquor by ion chromatography coupled with pulsed amperometric detection (IC-PAD). Eluent concentration, interferent evaluation and method performance were discussed. Results show that free cyanide in liquor can be rapidly determined by the optimised IC-PAD method. A sample requires only 1:100 dilution and simple filtration before being subjected to IC-PAD. The linear range is 1-5000 μg/L with an R value of 0.9998. The detection limit is 1 μg/L for a 25 μL injection loop. The overall relative standard deviation (RSD) of the method is less than 5%, and the recovery range is from 98.1% to 105.0%. This study has been proven significant and may have potential applications in liquors analysis.

  10. Automated determination of bromide in waters by ion chromatography with an amperometric detector

    Science.gov (United States)

    Pyen, G.S.; Erdmann, D.E.

    1983-01-01

    An automated ion chromatograph, including a program controller, an automatic sampler, an integrator, and an amperometric detector, was used to develop a procedure for the determination of bromide in rain water and many ground waters. Approximately 10 min is required to obtain a chromatogram. The detection limit for bromide is 0.01 mg l-1 and the relative standard deivation is <5% for bromide concentrations between 0.05 and 0.5 mg l-1. Chloride interferes if the chloride-to-bromide ratio is greater than 1 000:1 for a range of 0.01-0.1 mg l-1 bromide; similarly, chloride interferes in the 0.1-1.0 mg l-1 range if the ratio is greater than 5 000:1. In the latter case, a maximum of 2 000 mg l-1 of chloride can be tolerated. Recoveries of known concentrations of bromide added to several samples, ranged from 97 to 110%. ?? 1983.

  11. A CMOS detection chip for amperometric sensors with chopper stabilized incremental ΔΣ ADC

    Science.gov (United States)

    Min, Chen; Yuntao, Liu; Jingbo, Xiao; Jie, Chen

    2016-06-01

    This paper presents a low noise complimentary metal-oxide-semiconductor (CMOS) detection chip for amperometric electrochemical sensors. In order to effectively remove the input offset of the cascaded integrators and the low frequency noise in the modulator, a novel offset cancellation chopping scheme was proposed in the Incremental ΔΣ analog to digital converter (IADC). A novel low power potentiostat was employed in this chip to provide the biasing voltage for the sensor while mirroring the sensor current out for detection. The chip communicates with FPGA through standard built in I2C interface and SPI bus. Fabricated in 0.18-μm CMOS process, this chip detects current signal with high accuracy and high linearity. A prototype microsystem was produced to verify the detection chip performance with current input as well as micro-sensors. Project supported by the State Key Development Program for Basic Research of China (No. 2015CB352100).

  12. A CMOS analog front-end chip for amperometric electrochemical sensors

    Science.gov (United States)

    Zhichao, Li; Yuntao, Liu; Min, Chen; Jingbo, Xiao; Jie, Chen

    2015-07-01

    This paper reports a complimentary metal-oxide-semiconductor (CMOS) analog front-end chip for amperometric electrochemical sensors. The chip includes a digital configuration circuit, which can communicate with an external microcontroller by employing an I2C interface bus, and thus is highly programmable. Digital correlative double samples technique and an incremental sigma-delta analog to digital converter (Σ-Δ ADC) are employed to achieve a new proposed system architecture with double samples. The chip has been fabricated in a standard 0.18-μm CMOS process with high-precision and high-linearity performance occupying an area of 1.3 × 1.9 mm2. Sample solutions with various phosphate concentrations have been detected with a step concentration of 0.01 mg/L. Project supported by the National Key Basic Research and Development Project (No. 2015CB352103).

  13. Simultaneous topographic and amperometric membrane mapping using an AFM probe integrated biosensor.

    Science.gov (United States)

    Stanca, Sarmiza Elena; Csaki, Andrea; Urban, Matthias; Nietzsche, Sandor; Biskup, Christoph; Fritzsche, Wolfgang

    2011-02-15

    The investigation of the plasma membrane with intercorrelated multiparameter techniques is a prerequisite for understanding its function. Presented here, is a simultaneous electrochemical and topographic study of the cell membrane using a miniaturized amperometric enzymatic biosensor. The fabrication of this biosensor is also reported. The biosensor combines a scanning force microscopy (AFM) gold-coated cantilever and an enzymatic transducer layer of peroxidases (PODs). When these enzymes are brought in contact with the substrate, the specific redox reaction produces an electric current. The intensity of this current is detected simultaneously with the surface imaging. For sensor characterization, hydroquinone-2-carboxylic acid (HQ) is selected as an intrinsic source of H(2)O(2). HQ has been electrochemically regenerated by the reduction of antraquinone-2-carboxylic acid (AQ). The biosensor reaches the steady state value of the current intensity in 1 ± 0.2s.

  14. Analysis for urinary catecholamines by liquid chromatography with amperometric detection: methodology and clinical interpretation of results.

    Science.gov (United States)

    Moyer, T P; Jiang, N S; Tyce, G M; Sheps, S G

    1979-02-01

    A method is presented for the quantitative analysis of urinary unconjugated norepinephrine, epinephrine, and dopamine as discrete entities. The procedure requires initial purification of the specimen on aluminum oxide and a boric acid-gel. We used "high-performance" reversed-phase paired-ion chromatography, with a flow-through amperometric cell as the detector. The CV was 6% for determination of norepinephrine, 11% for epinephrine, and 6% for dopamine monitored at physiologic concentrations of these compounds in urine. In a population study, urine specimens from 117 normal pediatric and adult subjects, 85 hypertensive patients, and 22 patients with surgically proved pheochromocytoma were analyzed. The specificity of the method for detection of pheochromocytoma was 100%, with a sensitivity of 97%.

  15. An amperometric enzyme biosensor for real-time measurements of cellobiohydrolase activity on insoluble cellulose

    DEFF Research Database (Denmark)

    Cruys-Bagger, Nicolaj; Guilin, Ren; Tatsumi, Hirosuke;

    2012-01-01

    An amperometric enzyme biosensor for continuous detection of cellobiose has been implemented as an enzyme assay for cellulases. We show that the initial kinetics for cellobiohydrolase I, Cel7A from Trichoderma reesei, acting on different types of cellulose substrates, semi-crystalline and amorphous......) and this provided experimental access to the transient kinetics of cellobiohydrolases acting on insoluble cellulose. The response from the CDH-biosensor during enzymatic hydrolysis was corrected for the specificity of PcCDH for the β-anomer of cello-oligosaccharides and the approach were validated against HPLC....... It is suggested that quantitative, real-time data on pure insoluble cellulose substrates will be useful in attempts to probe the molecular mechanism underlying enzymatic hydrolysis of cellulose...

  16. Characterization of Hg(II) binding with different length phytochelatins using liquid chromatography and amperometric detection.

    Science.gov (United States)

    Dago, Angela; González-García, Olga; Ariño, Cristina; Díaz-Cruz, José Manuel; Esteban, Miquel

    2011-06-10

    A simple and rapid methodology is optimised to analyse mixtures of different phytochelatins (PC(n), n=2-5) with Hg(II) by HPLC with amperometric detection as a first step towards the analysis of extracts of plants stressed with Hg(II). The separation was achieved in a C(18) column with a mobile phase of 0.1% trifluoroacetic acid (TFA) in water and 0.1% TFA in acetonitrile using gradient elution. Electrochemical detection with glassy carbon electrode and UV-vis detection were used in series. This methodology can clearly distinguish between the free peptides and their complexes and permits to study the evolution of the different complexes formed and predicts the possible interactions between the long chain phytochelatin complexes. ESI-MS is used as a complementary technique to find out the stoichiometries of such long chain phytochelatin complexes.

  17. An Amperometric Biosensor for Uric Acid Determination Prepared From Uricase Immobilized in Polyaniline-Polypyrrole Film

    Directory of Open Access Journals (Sweden)

    Fatma Arslan

    2008-09-01

    Full Text Available A new amperometric uric acid biosensor was developed by immobilizing uricase by a glutaraldehyde crosslinking procedure on polyaniline-polypyrrole (pani-ppy composite film on the surface of a platinum electrode. Determination of uric acid was performed by the oxidation of enzymatically generated H2O2 at 0.4 V vs. Ag/AgCl. The linear working range of the biosensor was 2.5×10-6 – 8.5×10-5 M and the response time was about 70 s. The effects of pH, temperature were investigated and optimum parameters were found to be 9.0, 55 oC, respectively. The stability and reproducibility of the enzyme electrode have been also studied.

  18. Application of polyaniline/sol-gel derived tetraethylorthosilicate films to an amperometric lactate biosensor.

    Science.gov (United States)

    Chaubey, Asha; Pande, K K; Malhotra, B D

    2003-11-01

    The electrochemical entrapment of polyaniline (PANI) onto sol-gel derived tetraethylorthosilicate (TEOS) films deposited onto indium-tin-oxide (ITO) coated glass has been utilized for immobilization of lactate dehydrogenase (LDH). The performance of these sol-gel/PANI/LDH electrodes has been investigated as a function of the lactate concentration, applied potential, pH of the medium and interferents. The amperometric response of the electrodes under optimum conditions exhibited a linear relationship from 1 mM to 4 mM. An attempt has been made to extend the linearity up to 10 mM for lactate by coating an external layer of polyvinyl chloride (PVC) over the sol-gel/PANI/LDH electrodes with a correlation coefficient of 0.89. These sol-gel/PANI/LDH electrodes have a response time of about 60 s, a shelf life of about 8 weeks at 0-4 degrees C and have implications in a lactate biosensor.

  19. 一种新型唾液sIgA电流型纳米免疫生物传感器的制备%Preparation of a novel salivary sIgA amperometric nano-immunosensor

    Institute of Scientific and Technical Information of China (English)

    王艳; 甄生航; 郑军; 马霄; 刘成桂; 朱洋

    2010-01-01

    通过层层自组装,将硫堇(Thi)和纳米金(GNPs)修饰到Nation修饰的玻碳(GCE)电极表面,利用纳米金单层吸附唾液分泌性免疫球蛋白A(sIgA),最后用辣根过氧化物酶(HRP)封闭电极上的非特异性吸附位点,构建了一种检测唾液sIgA的新型电流型纳米免疫生物传感器.该生物传感器灵敏度高,特异性好,测试方便,检测线性范围为6.5-300mg/L,检出限为3.0mg/L;电流值达到95%稳态时间小于20s.探讨了抗体和底物浓度,pH值和温度,孵育时间,干扰物对传感器的影响.该传感器与ELISA法相关性良好(R=0.98932,P<0.001),可用于唾液sIgA的快速、准确检测,从而判断人体局部免疫状况.

  20. Development of an amperometric biosensing method for the determination of L-fucose in pretreated urine.

    Science.gov (United States)

    Tsiafoulis, Constantinos G; Prodromidis, Mamas I; Karayannis, Miltiades I

    2004-10-15

    The first amperometric biosensing method for the determination of L-fucose is described. L-Fucose is the objective of much current research, as it is considered as a potential marker for various pathologic disorders. Recombinant L-fucose dehydrogenase, having as cofactor beta-nicotinamide adenine dinucleotide phosphate (NAD+P), was cross-linked in a water-soluble photosensitive polymer matrix, that is, polyvinyl alcohol (PVA) modified with styrylpyridinium (SbQ), in the presence of BSA and glutaraldehyde. The resulting membrane was sandwiched between two polycarbonate membranes and was mounted in an amperometric cell. The oxidation of the enzymatically produced NADPH was monitored at a platinum anode at +0.25 V versus a silver pseudoreference electrode in the presence of ferricyanide. The system was fully optimized with respect to various analytical parameters. Regarding to the mechanical properties of the membrane and the storage stability of the immobilized enzyme, various parameters were also optimized. Several methods for the pretreatment of urine samples were investigated. Treatment of the samples with PbO2 found to eliminate the interference effect of various electroactive species exist in urine; optimum incubation time was determined since at prolonged incubation times L-fucose is also affected. Calibration curves for the direct and the mediated monitoring of NADPH were liner over the concentration ranges 0.04-1.0 mM (r2=0.9995) and 0.03-3.0 mM (r2=0.9997) fucose, respectively. The detection limits (S/N 3) were 2 and 1.5 microM fucose, respectively. The R.S.D. of the mediated biosensor is better than 1.5% (n=10, 0.5 mM fucose). The proposed biosensor correlates well with a reference enzymatic method and exhibits very good working and storage stability.

  1. Diagnostic tests for hepatitis C: recent trends in electrochemical immunosensor and genosensor analysis.

    Science.gov (United States)

    Uliana, Carolina V; Riccardi, Carla S; Yamanaka, Hideko

    2014-11-14

    Hepatitis C is a liver disease that is transmitted through contact with the blood of an infected person. An estimated 150 million individuals worldwide have been chronically infected with the hepatitis C virus (HCV). Hepatitis C shows significant genetic variation in the global population, due to the high rate of viral RNA mutation. There are six variants of the virus (HCV genotypes 1, 2, 3, 4, 5, and 6), with 15 recorded subtypes that vary in prevalence across different regions of the world. A variety of devices are used to diagnose hepatitis C, including HCV antibody test, HCV viral load test, HCV genotype test and liver biopsy. Rapid, inexpensive, sensitive, and robust analytical devices are therefore essential for effective diagnosis and monitoring of disease treatment. This review provides an overview of current electrochemical immunosensor and genosensor technologies employed in HCV detection. There are a limited number of publications showing electrochemical biosensors being used for the detection of HCV. Due to their simplicity, specificity, and reliability, electrochemical biosensor devices have potential clinical applications in several viral infections.

  2. Gold-coated carbon nanotube electrode arrays: Immunosensors for impedimetric detection of bone biomarkers.

    Science.gov (United States)

    Ramanathan, Madhumati; Patil, Mitali; Epur, Rigved; Yun, Yeoheung; Shanov, Vasselin; Schulz, Mark; Heineman, William R; Datta, Moni K; Kumta, Prashant N

    2016-03-15

    C-terminal telopeptide (cTx), a fragment generated during collagen degradation, is a key biomarker of bone resorption during the bone remodeling process. The presence of varying levels of cTx in the bloodstream can hence be indicative of abnormal bone metabolism. This study focuses on the development of an immunosensor utilizing carbon nanotube (CNT) electrodes coated with gold nanoparticles for the detection of cTx, which could ultimately lead to the development of an inexpensive and rapid point-of-care (POC) tool for bone metabolism detection and prognostics. Electrochemical impedance spectroscopy (EIS) was implemented to monitor and detect the antigen-antibody binding events occurring on the surface of the gold-deposited CNT electrode. Type I cTx was used as the model protein to test the developed sensor. The sensor was accordingly characterized at various stages of development for evaluation of the optimal sensor performance. The biosensor could detect cTx levels as low as 0.05 ng/mL. The feasibility of the sensor for point-of-care (POC) applications was further demonstrated by determining the single frequency showing maximum changes in impedance, which was determined to be 18.75 Hz.

  3. Modification of alumina matrices through chemical etching and electroless deposition of nano-Au array for amperometric sensing

    Directory of Open Access Journals (Sweden)

    Valinčius Gintaras

    2007-01-01

    Full Text Available AbstractSimple nanoporous alumina matrix modification procedure, in which the electrically highly insulating alumina barrier layer at the bottom of the pores is replaced with the conductive layer of the gold beds, was described. This modification makes possible the direct electron exchange between the underlying aluminum support and the redox species encapsulated in the alumina pores, thus, providing the generic platform for the nanoporous alumina sensors (biosensors with the direct amperometric signal readout fabrication.

  4. Characterization of Hardwood-Derived Carboxymethylcellulose by High pH Anion Chromatography Using Pulsed Amperometric Detection

    OpenAIRE

    2010-01-01

    An approach for the quantitative analysis of substituent distribution in carboxymethylcellulose (CMC) is presented. In short, the high-pH anion-exchange chromatography method, coupled to pulsed amperometric detection (PAD), is introduced. Each of the seven derivatives in CMC is presented by a single peak on the PAD trace, thus enabling an easy quantification. New inside information on monomer composition is obtained by this novel method, which is essential for understanding the structure vers...

  5. Facile fabrication of an ultrasensitive sandwich-type electrochemical immunosensor for the quantitative detection of alpha fetoprotein using multifunctional mesoporous silica as platform and label for signal amplification.

    Science.gov (United States)

    Wang, Yulan; Li, Xiaojian; Cao, Wei; Li, Yueyun; Li, He; Du, Bin; Wei, Qin

    2014-11-01

    A novel and ultrasensitive sandwich-type electrochemical immunosensor was designed for the quantitative detection of alpha fetoprotein (AFP) using multifunctional mesoporous silica (MCM-41) as platform and label for signal amplification. MCM-41 has high specific surface area, high pore volume, large density of surface silanol groups (SiOH) and good biocompatibility. MCM-41 functionalized with 3-aminopropyltriethoxysilane (APTES), gold nanoparticles (Au NPs) and toluidine blue (TB) could enhance electrochemical signals. Moreover, primary antibodies (Ab1) and secondary antibodies (Ab2) could be effectively immobilized onto the multifunctional MCM-41 by the interaction between Au NPs and amino groups (-NH2) on antibodies. Using multifunctional MCM-41 as a platform and label could greatly simplify the fabrication process and result in a high sensitivity of the designed immunosensor. Under optimal conditions, the designed immunosensor exhibited a wide liner range from 10(-4) ng/mL to 10(3) ng/mL with a low detection limit of 0.05 pg/mL for AFP. The designed immunosensor showed acceptable selectivity, reproducibility and stability, which could provide potential applications in clinical monitoring of AFP.

  6. Glucose oxidase-modified carbon-felt-reactor coupled with peroxidase-modified carbon-felt-detector for amperometric flow determination of glucose

    Energy Technology Data Exchange (ETDEWEB)

    Wang Yue [School of Chemical Engineering, University of Science and Technology LiaoNing, 185 Qianshan Middle Road, High-tech Zone, Anshan, LiaoNing, 114501 (China); Hasebe, Yasushi, E-mail: hasebe@sit.ac.jp [Department of Life Science and Green Chemistry, Faculty of Engineering, Saitama Institute of Technology, 1690, Fusaiji, Fukaya, Saitama 369-0293 (Japan)

    2012-04-01

    Glucose oxidase (GOx) and horseradish peroxidase (HRP) were covalently immobilized on a porous carbon-felt (CF) by using cyanuric chloride (CC) as a linking reagent. The resulting GOx-modified-CF (GOx-ccCF) was used as column-type enzyme reactor and placed on upstream of the HRP-ccCF-based H{sub 2}O{sub 2} flow-detector to fabricate amperometric flow-biosensor for glucose. Sensor setting conditions and the operational conditions were optimized, and the analytical performance characteristics of the resulting flow-biosensor were evaluated. The chemical modification of the GOx via CC was found to be effective to obtain larger catalytic activity as compared with the physical adsorption. Under the optimized conditions (i.e., volume ratio of the GOx-ccCF-reactor to the HRP-ccCF-detector is 1.0; applied potential is - 0.12 V vs. Ag/AgCl; carrier pH is 6.5; and carrier flow rate is 4.3 ml/min), highly selective and quite reproducible peak current responses toward glucose were obtained: the RSD for 30 consecutive injections of 3 mM glucose was 1.04%, and no serious interferences were observed for fructose, ethanol, uric acid, urea and tartaric acid for the amperometric measurements of glucose. The magnitude of the cathodic peak currents for glucose was linear up to 5 mM (sensitivity, 6.38 {+-} 0.32 {mu}A/{mu}M) with the limit detection of 9.4 {mu}M (S/N = 3, noise level, 20 nA). The present GOx-ccCF-reactor and HRP-ccCF-detector-coupled flow-glucose biosensor was utilized for the determination of glucose in beverages and liquors, and the analytical results by the sensor were in fairly good agreement with those by the conventional spectrophotometry. - Highlights: Black-Right-Pointing-Pointer Glucose oxidase (GOx) and peroxidase (HRP) were modified on carbon-felt. Black-Right-Pointing-Pointer GOx-CF reactor and HRP-CF detector-coupled flow glucose biosensor was developed. Black-Right-Pointing-Pointer This flow biosensor enabled the determination of glucose in beverages and

  7. Improvement of Accuracy in Flow Immunosensor System by Introduction of Poly-2-[3-(methacryloylaminopropylammonio]ethyl 3-aminopropyl Phosphate

    Directory of Open Access Journals (Sweden)

    Yusuke Fuchiwaki

    2011-01-01

    Full Text Available In order to improve the accuracy of immunosensor systems, poly-2-[3-(methacryloylaminopropylammonio]ethyl 3-aminopropyl phosphate (poly-3MAm3AP, which includes both phosphorylcholine and amino groups, was synthesized and applied to the preparation of antibody-immobilized beads. Acting as an antibody-immobilizing material, poly-3MAm3AP is expected to significantly lower nonspecific adsorption due to the presence of the phosphorylcholine group and recognize large numbers of analytes due to the increase in antibody-immobilizing sites. The elimination of nonspecific adsorption was compared between the formation of a blocking layer on antibody-immobilized beads and the introduction of a material to combine antibody with beads. Determination with specific and nonspecific antibodies was then investigated for the estimation of signal-to-noise ratio. Signal intensities with superior signal-to-noise ratios were obtained when poly-3MAm3AP was introduced. This may be due to the increase in antibody-immobilizing sites and the extended space for antigen-antibody interaction resulting from the electrostatic repulsion of poly-3MAm3AP. Thus, the application of poly-3MAm3AP coatings to immunoassay beads was able to improve the accuracy of flow immunosensor systems.

  8. An ultrasensitive sandwich type electrochemiluminescence immunosensor for triiodothyronine detection using silver nanoparticle-decorated graphene oxide as a nanocarrier.

    Science.gov (United States)

    Chou, Hung-Tao; Fu, Chien-Yu; Lee, Chi-Young; Tai, Nyan-Hwa; Chang, Hwan-You

    2015-09-15

    An ultrasensitive electrochemiluminescence (ECL) immunosensor was constructed to detect 3,3',5-triiodothyronine (T3). The system employed T3-conjugated, silver nanoparticle-decorated carboxylic graphene oxide (Ag@fGO-T3) as a carrier and anti-T3 antibody-tris(2,2'-bipyridyl) ruthenium(II) (Ru(bpy)3(2+)) as a probe. The Ag@fGO-T3 and Ru(bpy)3(2+) complex could be mobilized rapidly to the anode in the reaction chamber through electrophoresis. The fGO is reduced electrochemically at the electrode, and the electrons could transfer from an anode to the Ru(bpy)3(2+). The complex is excited at the electrode and an ECL signal is produced upon reacting with tripropylamine (TPrA). Because of its large surface area and excellent conductivity, Ag@fGO could enhance ECL signal significantly in the system. Quantitative measurement of T3 could be achieved in the range from 0.1 pg/mL to 0.8 ng/mL with a detection limit of 0.05 pg/mL. In addition, the novel immunosensor showed good specificity in the presence of serum, indicating its high potential in clinical use.

  9. Immobilization strategy for enhancing sensitivity of immunosensors: L-Asparagine-AuNPs as a promising alternative of EDC-NHS activated citrate-AuNPs for antibody immobilization.

    Science.gov (United States)

    Raghav, Ragini; Srivastava, Sudha

    2016-04-15

    This paper addresses the question - Is EDC-NHS activated gold nanoparticles modified electrode surface the best available option for antibody immobilization for immunosensor fabrication? Is there any other alternative covalent immobilization strategy for orthogonal orientation of antibody, ensuring enhanced sensitivity of immunosensors? Does EDC-NHS activation of carboxyl functionalized nanoparticles surface really leads to orthogonal or directed immobilization of antibody? Gold nanoparticles synthesized using L-Asparagine as reducing and stabilization agent were employed for orthogonal immobilization of antibody for immunosensor fabrication. Anti-CA125 antibody was used as a model system for immunosensor fabrication. A comparative evaluation of immunosensors fabricated using L-Asparagine stabilized gold nanoparticles and citrate stabilized gold nanoparticles via different immobilization strategies/chemistries was done. The three strategies involved immobilization of Anti-CA125 antibody - (1) after EDC-NHS activation of citrate stabilized gold nanoparticles, (2) directly onto citrate stabilized gold nanoparticles and (3) directly onto L-Asparagine stabilized gold nanoparticles modified electrode surfaces. Comparative evaluation of Impedimetric response characteristics showed 2.5 times increase in sensitivity (349.36 Ω/(IU/mL)/cm(2)) in case of third strategy as compared to first (147.53 Ω/(IU/mL)/cm(2)) and twice that of second strategy (166.24 Ω/(IU/mL)/cm(2)). Additionally, an extended dynamic range of 0-750 IU/mL was observed while for others it was up to 500 IU/mL. Amino acid coated gold nanoparticles ensured orthogonal immobilization, lesser randomization, with 88% of active antibody available for antigen binding as opposed to other two strategies with less than 30% active antibody.

  10. Amperometric biosensor for the determination of phenols using a crude extract of sweet potato

    Energy Technology Data Exchange (ETDEWEB)

    Cruz Vieira, I. da; Fatibello-Filho, O. [Universidade Federal de Sa Carlos (Brazil)

    1997-03-01

    An amperometric biosensor for the determination of phenols is proposed using a crude extract of sweet potato (Ipomoea batatas (L.) Lam.) as an enzymatic source of polyphenol oxidase (PPO; tyrosinase; catechol oxidase; EC 1.14.18.1). The biosensor is constructed by the immobilization of sweet potato crude extract with glutaraldehyde and bovine serum albumin onto an oxygen membrane. This biosensor provides a linear response for catechol, pyrogallol, phenol and p-cresol in the concentration ranges of 2.0 x 10{sup -5} -4.3 x 10{sup -4} mol L{sup -1}, 2.0 x 10{sup -5} -4.3 x 10{sup -4} mol L{sup -1}, 2.0 x 10{sup -5} -4.5 x 10{sup -4} mol L{sup -1} and 2.0 x 10{sup -5} -4.5 x 10{sup -4} mol L{sup -1}, respectively. The response time was about 3-5 min for the useful response range, and the lifetime of this electrode was excellent for fifteen days (over 220 determinations for each enzymatic membrane). Application of this biosensor for the determination of phenols in industrial wastewaters is presented.

  11. Electrocatalytic amperometric determination of amitrole using a cobalt-phthalocyanine-modified carbon paste electrode.

    Science.gov (United States)

    Chicharro, Manuel; Zapardiel, Antonio; Bermejo, Esperanza; Moreno, Mónica; Madrid, Elena

    2002-07-01

    Cobalt-phthalocyanine-modified carbon paste electrodes are shown to be excellent indicators for electrocatalytic amperometric measurements of triazolic herbicides such as amitrole, at low oxidation potentials (+0.40 V). The detection and determination of amitrole in flow injection analysis with a modified carbon paste electrode with Co-phthalocyanine is described. The concentrations of amitrole in 0.1 M NaOH solutions were determined using the electrocatalytic oxidation signal corresponding to the Co(II)/Co(III) redox process. A detection limit of 0.04 microg mL(-1) (4 ng amitrole) was obtained for a sample loop of 100 microL at a fixed potential of +0.55 V (vs. Ag/AgCl) in 0.1 M NaOH and a flow rate of 4.0 mL min(-1). Furthermore, the modified carbon paste electrodes offers reproducible responses in such a system, and the relative standard deviation was 3.3% using the same surface, 5.1% using different surface, and 6.9% using different pastes. The performance of the cobalt-phthalocyanine-modified carbon paste electrodes is illustrated here for the determination of amitrole in commercial formulations. The response of the electrodes is stable, with more than 80% of the initial retained activity after 50 min of continuous use.

  12. CMOS Amperometric ADC With High Sensitivity, Dynamic Range and Power Efficiency for Air Quality Monitoring.

    Science.gov (United States)

    Li, Haitao; Boling, C Sam; Mason, Andrew J

    2016-08-01

    Airborne pollutants are a leading cause of illness and mortality globally. Electrochemical gas sensors show great promise for personal air quality monitoring to address this worldwide health crisis. However, implementing miniaturized arrays of such sensors demands high performance instrumentation circuits that simultaneously meet challenging power, area, sensitivity, noise and dynamic range goals. This paper presents a new multi-channel CMOS amperometric ADC featuring pixel-level architecture for gas sensor arrays. The circuit combines digital modulation of input currents and an incremental Σ∆ ADC to achieve wide dynamic range and high sensitivity with very high power efficiency and compact size. Fabricated in 0.5 [Formula: see text] CMOS, the circuit was measured to have 164 dB cross-scale dynamic range, 100 fA sensitivity while consuming only 241 [Formula: see text] and 0.157 [Formula: see text] active area per channel. Electrochemical experiments with liquid and gas targets demonstrate the circuit's real-time response to a wide range of analyte concentrations.

  13. Characterisation of brewpub beer carbohydrates using high performance anion exchange chromatography coupled with pulsed amperometric detection.

    Science.gov (United States)

    Arfelli, Giuseppe; Sartini, Elisa

    2014-01-01

    High performance anion exchange chromatography (HPAEC) coupled with pulsed amperometric detection (PAD) was optimised in order to quantify mannose, maltose, maltotriose, maltotetraose, maltopentaose, maltohexaose and maltoheptaose content of beer. The method allows the determination of above mentioned oligosaccharides, in a single chromatographic run, without any pre-treatment. Limit of detection and limit of quantification were suitable for beer. Accuracy and repeatability were good for the entire amount considered. Once optimised HPAEC PAD for the specific matrix, the second goal of this research was to verify the possibility to discriminate beers, depending on their style. The carbohydrates content of brewpub commercial beers was very variable, ranging from 19.3 to 1469mg/L (mannose), 34.5 to 2882mg/L (maltose), 141.9 to 20731mg/L (maltotriose), 168.5 to 7650mg/L (maltotetraose), 20.1 to 2537mg/L (maltopentaose), 22.9 to 3295mg/L (maltohexaose), 8.5 to 2492mg/L (maltoeptaose), even in the same style of beer. However, the carbohydrates content was useful, jointed with other compounds amount, to discriminate different styles of beer. As a matter of fact, principal component analysis put in evidence beer differences considering some fermentation conditions and colour.

  14. Analysis of Trace Ingredients in Green Tea by Capillary Electrophoresis with Amperometric Detection

    Institute of Scientific and Technical Information of China (English)

    LI Ping; DONG Shu-Qing; WANG Qing-Jiang; FANG Yu-Zhi

    2008-01-01

    In this paper, four trace ingredients (rutin, gallic acid, quercetin, chlorogenic acid) in green tea were simultaneously determined by capillary electrophoresis coupled with amperometric detection (CE-AD). Effects of several important factors such as the pH and concentration of running buffer, separation voltage, injection time and detection potential were investigated to acquire the optimum conditions. Under the optimum conditions, the analytes could be separated within 20 min at a separation voltage of 18 kV in a 60 mmol/L borate buffer (pH 8.7). A 300 μmdiameter carbon disk electrode generated good responses at 950 mV (vs. SCE) for all analytes. The relationship between the peak currents and concentrations of the analytes was linear over about three orders of magnitude with demonstrated long-term stability and reproducibility with relative standard deviations less than 3% for both migration time and peak current (n=7), which could be successfully used for the determination of the analytes in green tea with satisfactory assay results.

  15. Amperometric detection of lactose using β-galactosidase immobilized in layer-by-layer films.

    Science.gov (United States)

    Campos, Paula P; Moraes, Marli L; Volpati, Diogo; Miranda, Paulo B; Oliveira, Osvaldo N; Ferreira, Marystela

    2014-07-23

    A direct, low-cost method to determine the concentration of lactose is an important goal with possible impact in various types of industry. In this study, a biosensor is reported that exploits the specific interaction between lactose and the enzyme β-galactosidase (β-Gal) normally employed to process lactose into glucose and galactose for lactose-intolerant people. The biosensor was made with β-Gal immobilized in layer-by-layer (LbL) films with the polyelectrolyte poly(ethylene imine) (PEI) and poly(vinyl sufonate) (PVS) on an indium tin oxide (ITO) electrode modified with a layer of Prussian Blue (PB). With an ITO/PB/(PEI/PVS)1(PEI/β-Gal)30 architecture, lactose could be determined with an amperometric method with sensitivity of 0.31 μA mmol(-1) cm(-2) and detection limit of 1.13 mmol L(-1), which is sufficient for detecting lactose in milk and for clinical exams. Detection occurred via a cascade reaction involving glucose oxidase titrated as electrolytic solution in the electrochemical cell, while PB allowed for operation at 0.0 V versus saturated calomel electrode, thus avoiding effects from interfering species. Sum-frequency generation spectroscopy data for the interface between the LbL film and a buffer containing lactose indicated that β-Gal lost order, which is the first demonstration of structural effects induced by the molecular recognition interaction with lactose.

  16. Amperometric Determination of Sulfite by Gas Diffusion- Sequential Injection with Boron-Doped Diamond Electrode

    Directory of Open Access Journals (Sweden)

    Orawon Chailapakul

    2008-03-01

    Full Text Available A gas diffusion sequential injection system with amperometric detection using aboron-doped diamond electrode was developed for the determination of sulfite. A gasdiffusion unit (GDU was used to prevent interference from sample matrices for theelectrochemical measurement. The sample was mixed with an acid solution to generategaseous sulfur dioxide prior to its passage through the donor channel of the GDU. Thesulfur dioxide diffused through the PTFE hydrophobic membrane into a carrier solution of 0.1 M phosphate buffer (pH 8/0.1% sodium dodecyl sulfate in the acceptor channel of theGDU and turned to sulfite. Then the sulfite was carried to the electrochemical flow cell anddetected directly by amperometry using the boron-doped diamond electrode at 0.95 V(versus Ag/AgCl. Sodium dodecyl sulfate was added to the carrier solution to preventelectrode fouling. This method was applicable in the concentration range of 0.2-20 mgSO32−/L and a detection limit (S/N = 3 of 0.05 mg SO32−/L was achieved. This method wassuccessfully applied to the determination of sulfite in wines and the analytical resultsagreed well with those obtained by iodimetric titration. The relative standard deviations forthe analysis of sulfite in wines were in the range of 1.0-4.1 %. The sampling frequency was65 h−1.

  17. Short communication: Quantification of carbohydrates in whey permeate products using high-performance anion-exchange chromatography with pulsed amperometric detection.

    Science.gov (United States)

    Lee, Hyeyoung; de MeloSilva, Vitor Luiz; Liu, Yan; Barile, Daniela

    2015-11-01

    A method was developed for the characterization and quantification of the disaccharide lactose and 3 major bovine milk oligosaccharides (BMO) in dairy streams. Based on high-performance anion-exchange chromatography-pulsed amperometric detection (HPAE-PAD), this method is advantageous because it requires minimal sample preparation and achieves good chromatographic separation of oligosaccharide isomers within 30min. The linear dynamic range and limit of detection were 0.1 to 10mg/L and 0.03 to 0.22mg/L, respectively. Mean recoveries of the BMO were excellent and ranged from 98.4 to 100.4%. Without complicated sample preparation procedures, this HPAE-PAD method measured BMO [3'-sialyllactose (3'SL), 6'-sialyllactose (6'SL), and 6'-sialyllactosamine (6'SLN)] and lactose using a single instrument, therefore increasing the accuracy of the measurement and applicability for the dairy industry. In colostrum whey permeate, 3'SL, 6'SL, and 6'SLN were 94, 29, and 46mg/L, respectively. This work is the first to demonstrate that some commercial products, currently marketed for supporting a healthy immune system, contain significant amounts of bioactive BMO and therefore, carry additional bioactivities.

  18. Carbon nanotubes and graphene modified screen-printed carbon electrodes as sensitive sensors for the determination of phytochelatins in plants using liquid chromatography with amperometric detection.

    Science.gov (United States)

    Dago, Àngela; Navarro, Javier; Ariño, Cristina; Díaz-Cruz, José Manuel; Esteban, Miquel

    2015-08-28

    Nanomaterials are of great interest for the development of electrochemical sensors. Multi-walled carbon nanotubes and graphene were used to modify the working electrode surface of different screen-printed carbon electrodes (SPCE) with the aim of improving the sensitivity of the SPCE and comparing it with the conventional glassy carbon electrode. To assay the usability of these sensors, a HPLC methodology with amperometric detection was developed to analyze several phytochelatins in plants of Hordeum vulgare and Glycine max treated with Hg(II) or Cd(II) giving detection limits in the low μmolL(-1) range. Phytochelatins are low molecular weight peptides with the general structure γ-(Glu-Cys)n-Gly (n=2-5) which are synthesized in plants in the presence of heavy metal ions. These compounds can chelate heavy metal ions by the formation of complexes which, are transported to the vacuoles, where the toxicity is not threatening. For this reason phytochelatins are essential in the detoxification of heavy metal ions in plants. The developed HPLC method uses a mobile phase of 1% of formic acid in water with KNO3 or NaCl (pH=2.00) and 1% of formic acid in acetonitrile. Electrochemical detection at different carbon-based electrodes was used. Among the sensors tested, the conventional glassy carbon electrode offers the best sensitivity although modification improves the sensitivity of the SPCE. Glutathione and several isoforms of phytochelatin two were found in plant extracts of both studied species.

  19. Biomolecule-based formaldehyde resin microspheres loaded with Au nanoparticles: a novel immunoassay for detection of tumor markers in human serum.

    Science.gov (United States)

    Lu, Wenbo; Qian, Chen; Bi, Liyan; Tao, Lin; Ge, Juan; Dong, Jian; Qian, Weiping

    2014-03-15

    A surfactant-free and template-free method for the high-yield synthesis of biomolecule (serotonin)-based formaldehyde resin (BFR) microspheres is proposed for the first time. The colloidal microspheres loaded with Au nanoparticles (AuNPs) prepared by a convenient in-situ synthesis of AuNPs on BFR (AuNPs/BFR) microsphere surface show good stability. AuNPs/BFR microspheres not only favor the immobilization of antibody but also facilitate the electron transfer. It is found that the resultant AuNPs/BFR microspheres can be designed to act as a sensitive label-free electrochemical immunosensor for carcinoembryonic antigen (CEA) determination. The immunosensor is prepared by immobilizing capture anti-CEA on AuNPs/BFR microspheres assembled on thionine (TH) modified glassy carbon electrode (GCE). TH acts as the redox probe. Under the optimized conditions, the linear range of the proposed immunosensor is estimated to be from 25 pg/mL to 2000 pg/mL (R=0.998) and the detection limit is estimated to be 3.5 pg/mL at a signal-to-noise ratio of 3. The prepared immunosensor for detection of CEA shows high sensitivity, reproducibility and stability. Our study demonstrates that the immunosensor can be used for the CEA detection in humans serum.

  20. Critical Evaluation of Acetylcholine Determination in Rat Brain Microdialysates using Ion-Pair Liquid Chromatography with Amperometric Detection

    Directory of Open Access Journals (Sweden)

    Yvette Michotte

    2008-08-01

    Full Text Available Liquid chromatography with amperometric detection remains the most widely used method for acetylcholine quantification in microdialysis samples. Separation of acetylcholine from choline and other matrix components on a microbore chromatographic column (1 mm internal diameter, conversion of acetylcholine in an immobilized enzyme reactor and detection of the produced hydrogen peroxide on a horseradish peroxidase redox polymer coated glassy carbon electrode, achieves sufficient sensitivity for acetylcholine quantification in rat brain microdialysates. However, a thourough validation within the concentration range required for this application has not been carried out before. Furthermore, a rapid degradation of the chromatographic columns and enzyme systems have been reported. In the present study an ion-pair liquid chromatography assay with amperometric detection was validated and its long-term stability evaluated. Working at pH 6.5 dramatically increased chromatographic stability without a loss in sensitivity compared to higher pH values. The lower limit of quantification of the method was 0.3 nM. At this concentration the repeatability was 15.7%, the inter-day precision 8.7% and the accuracy 103.6%. The chromatographic column was stable over 4 months, the immobilized enzyme reactor up to 2-3 months and the enzyme coating of the amperometric detector up to 1-2 months. The concentration of acetylcholine in 30 μl microdialysates obtained under basal conditions from the hippocampus of freely moving rats was 0.40 ± 0.12 nM (mean ± SD, n = 30. The present method is therefore suitable for acetylcholine determination in rat brain microdialysates.

  1. Ultrasensitive electrochemiluminescence immunoassay for tumor marker based on quantum dots coated carbon nanospheres

    Energy Technology Data Exchange (ETDEWEB)

    Li, Long; Zhang, Yan; Li, Shuai; Wang, Xiu; Li, Chen [Key Laboratory of Chemical Sensing and Analysis in Universities of Shandong, School of Chemistry and Chemical Engineering, University of Jinan, Jinan 250022 (China); Ge, Shenguang [Shandong Provincial Key Laboratory of Fluorine Chemistry and Chemical Materials, University of Jinan, Jinan 250022 (China); Yu, Jinghua, E-mail: ujn.yujh@gmail.com [Key Laboratory of Chemical Sensing and Analysis in Universities of Shandong, School of Chemistry and Chemical Engineering, University of Jinan, Jinan 250022 (China); Yan, Mei [Key Laboratory of Chemical Sensing and Analysis in Universities of Shandong, School of Chemistry and Chemical Engineering, University of Jinan, Jinan 250022 (China); Song, Xianrang [Cancer Research Center, Shandong Tumor Hospital, Jinan 250012 (China)

    2013-12-15

    In this work, a novel electrochemiluminescence (ECL) immunosensor based on CdTe quantum dots (QDs) coated carbon nanosphere (CN/QDs) for the detection of carcinoembryonic antigen (CEA) was developed. The carbon nanospheres (CNs) with good monodispersity and uniform structure were synthetized by a hydrothermal method using glucose as raw material. Then QDs functionized CNs were prepared and employed for signal amplification to improve the sensitivity and the detection limit of immunosensor. For this proposed immunosensor, chitosan was firstly deposited on the pretreated indium tin oxide (ITO) electrode surface, which promoted the electron transfer. Subsequently, gold nanoparticles (AuNPs) were assembled onto chitosan film modified electrode to improve the absorption capacity of antibodies. Then, primary antibodies were immobilized onto the electrode through the reaction between AuNPs and amino. At last bovine serum albumin (BSA) was employed to block the nonspecific binding sites. As a result, a novel ECL immunosensor was obtained on the prepared CN/QDs. The CEA was determined in the range of 0.005–200 ng mL{sup −1}, with a low detection limit of 1.2 pg mL{sup −1} (S/N=3). The proposed ECL immunosensor provides a rapid, simple, and sensitive immunoassay protocol for protein detection, which could be applied in more bioanalytical systems. -- Highlights: • A sandwich-type electrochemluminence immunosensor was fabricated. • CdTe quantum dots coated carbon nanospheres were used to amplify signals. • Au–chitosan biocompatible membrane modified on ITO electrode to capture antibodies.

  2. Separation of alditols of interest in food products by high-performance anion-exchange chromatography with pulsed amperometric detection.

    Science.gov (United States)

    Corradini, C; Canali, G; Cogliandro, E; Nicoletti, I

    1997-12-12

    High-performance anion-exchange chromatography (HPAEC)-pulsed amperometric detection (PAD) employing a CarboPac MA 1 column was investigated with respect to mobile phase composition, linear response characteristics, repeatability, reproducibility and sensitivity with different alditols used as sugar substitutes in food and confectionery products. The energy-reduced bulk sweeteners isomalt and maltitol were well resolved in less than 25 min by isocratic elution with 600 mM sodium hydroxide solution. HPAEC-PAD was also successfully applied to the determination of alditols in sugar-free products and a low-calorie sweetener containing sorbitol, mannitol and fructose at different levels.

  3. Synthesis of Cu2 O nanoparticle and amperometric detection of dopamine based on Cu2 O modified electrode%纳米Cu2 O材料的合成及其修饰电极安培法测定多巴胺

    Institute of Scientific and Technical Information of China (English)

    吴芳辉; 程立春; 陈乐; 宋超; 张奥亚; 梅洁

    2014-01-01

    In this work,a one-step,simple route towards synthesizing cuprous oxide( Cu2 O) nanoparticles with uniform particle size was explored using chemical reduction method. Then nanomaterial of Cu2 O was characterized by scanning electron microscopy ( SEM). In pH 7. 0 phosphate buffer solution,Cu2 O and Nafion complex modified electrode exhibited a strong electrocatalytic effect to dopamine( DA) compare with the bare glassy carbon electrode. Under the optimized experimental conditions,an electroanalytical system of trace dopamine was established based on chronoamperometry of electrocatalytic properties of Cu2 O modified electrode. The detection limit is 0. 17 μmol·L-1 with the sensitivity of 20. 44 μA mM-1 and the reaction time of less than 3 s in a concentration range of 0. 5~270 μmol·L-1 . The concentration of dopamine was able to be accurately measured at 20 times the AA coexistence due to increase Nafion film. The results of voltammetric determinations show a good reproducibility and stability,and the relative standard deviation ( RSD ) was 3. 3% for the slope of the calibration curve based on the 11 measurements of 50 μM dopamine. Moreover,the developed sensor could be successfully applied to determine dopamine in simulation preparations.%本文采用简单的一步化学还原方法合成了粒径均一的纳米Cu2 O材料并采用扫描电子显微镜对其形貌进行了表征。研究发现,在pH 7.0的磷酸盐缓冲溶液中,采用纳米Cu2 O和Nafion(全氟磺酸离子交换树脂)膜制备的复合修饰电极对多巴胺( DA)呈现出较强的电化学催化作用。优化实验条件后,建立了计时电流法直接测定多巴胺的痕量分析体系。在0.5~270μmol·L-1浓度范围内,多巴胺的阳极峰电流与浓度呈良好的线性关系(r=0.9980),检测限为0.17μmol·L-1,灵敏度为20.44μA mM-1且响应时间不超过3 s。该电极可有效屏蔽抗坏血酸( AA)的干扰,在20倍AA共存下

  4. A Novel Amperometric Hydrogen Peroxide Biosensor Based on Thionine- Carbon Nanotube Modified Glassy Carbon Electrode%基于硫堇/碳纳米管修饰电极的新型过氧化氢电化学传感器

    Institute of Scientific and Technical Information of China (English)

    邓春艳; 阳明辉

    2012-01-01

    Based on the synergic effect of multi-walled carbon nanotubes(CNTs) and thionine(Th) , a novel hydrogen peroxide electrode ( HRP/GA - Th/CNTs/GC ) was fabricated by immobilizing horseradish peroxidase( HRP) onto the thionine(Th) /multi-walled carbon nanolubes modified electrode via glutaradehyde( GA) cross-linking. The introduction of CNTs into the electrode is helpful for the further immobilization of Th and the catalysis to H2O2. Furthermore, Th immobilized on CNTs by electrostatic adsorption not only can transfer electrons between the electrode and the redox activity center of the enzyme, but also can enable amine group( NH2) in Th to provide convenience for the immobilization of HRP enzyme via glutaradehyde cross-linking. Additionally, effects of various ex-perimental parameters on H2O2 sensing, including applied potential, pH value and electroactive in-terferent, were investigated. At an optimal potential of - 0. 3 V, the current response of the biosen-sor in phosphate buffer( pH 7. 0) was linear with H2O2 concentration from 5 μmol· L-1 to 40 μmol . L-1 , with a good detection limit (0. 3μmol L-1), a short response time (within 5 s) and a good anti-interferent ability.%基于碳纳米管(CNTs)和硫堇(Th)的协同效应,将辣根过氧化物酶(HRP)通过戊二醛(GA)交联作用固定在硫堇( Th)/CNTs修饰电极上,构造了一种新型酶电极(HRP/GA -Th/CNTs/GC).CNTs静电吸附正电荷的Th,而Th不仅可以促进电极和酶的氧化还原活性中心之间的电子传递,而且能使CNTs氨基(-NH2)功能化,从而利于HRP的固定.基于HRP/GA - Th/CNTs/GC电极的过氧化氢传感器具有较好的传感性能,且检出限低(0.3μmol·L-1)、响应时间短(5s内)、抗干扰能力强.

  5. Amperometric determination of serum total cholesterol with nanoparticles of cholesterol esterase and cholesterol oxidase.

    Science.gov (United States)

    Aggarwal, V; Malik, J; Prashant, A; Jaiwal, P K; Pundir, C S

    2016-05-01

    We describe the preparation of glutaraldehyde cross-linked and functionalized cholesterol esterase nanoparticles (ChENPs) and cholesterol oxidase nanoparticles (ChOxNPs) aggregates and their co-immobilization onto Au electrode for improved amperometric determination of serum total cholesterol. Transmission electron microscope (TEM) images of ChENPs and ChOxNPs showed their spherical shape and average size of 35.40 and 56.97 nm, respectively. Scanning electron microscope (SEM) studies of Au electrode confirmed the co-immobilization of enzyme nanoparticles (ENPs). The biosensor exhibited optimal response at pH 5.5 and 40°C within 5 s when polarized at +0.25 V versus Ag/AgCl. The working/linear range of the biosensor was 10-700 mg/dl for cholesterol. The sensor showed high sensitivity and measured total cholesterol as low as 0.1 mg/dl. The biosensor was evaluated and employed for total cholesterol determination in sera of apparently healthy and diseased persons. The analytical recovery of added cholesterol was 90%, whereas the within-batch and between-batch coefficients of variation (CVs) were less than 2% and less than 3%. There was a good correlation (r = 0.99) between serum cholesterol values as measured by the standard enzymic colorimetric method and the current method. The initial activity of ENPs/working electrode was reduced by 50% during its regular use (200 times) over a period of 60 days when stored dry at 4°C.

  6. Development of flow injection method for indirect copper determination with amperometric detection in drinking water samples

    Directory of Open Access Journals (Sweden)

    Nikolić-Mandić Snežana

    2012-01-01

    Full Text Available A gas-diffusion flow injection method with amperometric detection for indirect copper determination on a silver electrode is developed. The flow through system is equipped with two injection valves and a gas-diffusion unit. In the first step, a signal of cyanide solution was recorded. In the following step a signal of cyanide in the presence of copper was measured. Interferences (Cd(II, Co(II, Ag(I, Ni(II, Fe(III, Hg(II and Zn(II were investigated and successfully removed. The calibration graph is linear in the range 1-90 μmol dm-3 of copper, correlation coefficient is 0.993, the regression equation is I = (0.0455±0.0015c + (0.4611±0.0671, I is relative signal decrease in μA and c is concentration in μmol dm-3. Relative standard deviation for six consecutive injections of 30 μmol dm-3 copper(II was 1.47 % and for 1 μmol dm-3 copper(II was 3.40 %. The detection limit, calculated as 3 s/m (where s is a standard deviation of nine measurement of a reagent blank and m is the slope of the calibration curve, was 0.32 μmol dm-3, which corresponds to 2.44 ng of copper(II (loop volume was 0.12 cm3. The method enables 60 analyses per hour and it was successfully applied on determination of copper in drinking water samples. [Acknowledgements. The authors acknowledge the grant from the Ministry of Education and Science of the Republic of Serbia, Project number 172051

  7. Nanostructured cupric oxide electrode: An alternative to amperometric detection of carbohydrates in anion-exchange chromatography.

    Science.gov (United States)

    Barragan, José T C; Kubota, Lauro T

    2016-02-01

    In this paper, a new and low cost copper/cupric oxide nanostructured electrode is presented as an alternative to the amperometric detection of carbohydrates in high-performance anion exchange chromatography. The modified copper electrodes were prepared by a simple and fast method which resulted in the obtainment of homogeneously distributed nanostructures adhered to the surface with controlled chemical nature. The results, when compared to conventional copper electrodes, exhibited considerable improvements in analytical results, including: 1) Better repeatability in consecutive glucose detections, in which the percent relative standard deviation improved from 15.1% to 0.279%. 2) Significant improvements in the stability of the baseline and a decrease of the stabilization time, going from several hours to approximately 15 min. 3) Considerable increase in the sensitivity towards glucose, from 5.02 nA min mg L(-1) to 25.5 nA min mg L(-1). 4) Improvements in the detectability with limits as low as 1.09 pmol. 5) Wide working range of concentrations (1 × 10(-2) to 1 × 10(4) mg L(-1)). 6) Good linearity with correlation coefficients greater than 0.998. 7) Possibility of detecting different molecules of carbohydrates (lactose, maltose, sucrose cellobiose, sorbitol, fructose, glucose, galactose, manose, arabitol, xylose, ribose and arabnose). In comparison to the electrode that is more employed for this type of application (gold electrode), the low cost, the possibility of detection at constant potential and the equivalent detection limits presented by the new electrode material introduced in this work emerge as characteristics that make this material a powerful alternative considering the detection of carbohydrates in anion exchange chromatography.

  8. Enhanced optical immunosensor based on surface plasmon resonance for determination of transferrin.

    Science.gov (United States)

    Liu, Xia; Sun, Ying; Song, Daqian; Zhang, Qinglin; Tian, Yuan; Zhang, Hanqi

    2006-01-15

    Wavelength modulation surface plasmon resonance biosensors (SPR) using colloidal Au nanoparticles and double-linker sensing membrane enhancement are reported for determination of transferrin. The 2-mercaptoethylamine (MEA) was immobilized on the biosensor surface with traditional amine coupling method. The interaction between colloidal Au nanoparticles and MEA was investigated. The anti-transferrin was immobilized on the biosensor surface prepared with staphylococcal protein A (SPA). The interaction of the antibody and antigen was monitored in real time. The good response was obtained in the concentration range 1-20, 0.1-20 and 0.05-20 microg/mL for directly immune assay, double-linker assay and colloidal Au-amplified assay. The result clearly demonstrates that these methods may obtain significantly enhancement of sensitivity for the wavelength modulation SPR biosensor.

  9. Electrical immunosensor based on a submicron-gap interdigitated electrode and gold enhancement.

    Science.gov (United States)

    Ahn, Junhyoung; Lee, Tae Han; Li, Taihua; Heo, Kwang; Hong, Seunghun; Ko, Jeongheon; Kim, Yongsam; Shin, Yong-Beom; Kim, Min-Gon

    2011-08-15

    We demonstrated that the detection of human interleukin 5 (IL5) with a higher sensitivity than the enzyme-linked immunosorbent assay (ELISA) was possible using mass-producible submicron-gap interdigitated electrodes (IDEs) combined with signal amplification by a gold nanoparticle (AuNP) and gold enhancement. IDEs, facing comb-shape electrodes, can act as simple and miniaturized devices for immunoassay. An IDE with a gap size of 400nm was fabricated by a stepper photolithography process and was applied for the immunoassay of human IL5. A biotinylated anti-human IL5 was immobilized on the streptavidin-modified IDE, and biotin-bovine serum albumin (BSA) and BSA were added sequentially to reduce non-specific binding between the streptavidin-immobilized IDE surface and other proteins. The immunoassay procedure included three main steps: the reaction of human IL5 to form antigen-antibody complexes, the binding of AuNP conjugation with an antibody against human IL5 for the sandwich immunoassay, and gold enhancement for electrical signal amplification. The measurement of electrical current at each step showed that the gold enhancement step was very critical in detection of the concentration of human IL5. Analysis by scanning electron microscope (SEM) showed that close to 1μm particles were formed from 10nm AuNP by the gold enhancement reaction using gold ions and hydroxylamine. Under optimized conditions, human IL5 could be analyzed at 1pgmL(-1) with a wide dynamic range (from 10(-3) to 100ngmL(-1) concentrations).

  10. A nanostructured cerium oxide film-based immunosensor for mycotoxin detection

    Science.gov (United States)

    Kaushik, Ajeet; Rathee Solanki, Pratima; Ansari, Anees Ahmad; Ahmad, Sharif; Dhar Malhotra, Bansi

    2009-02-01

    Rabbit-immunoglobulin antibodies (r-IgGs) and bovine serum albumin (BSA) have been immobilized onto sol-gel-derived nanostructured cerium oxide (nanoCeO2) film fabricated onto an indium-tin-oxide (ITO) coated glass plate to detect ochratoxin-A (OTA). Broad reflection planes obtained in x-ray diffraction (XRD) patterns reveal the formation of CeO2 nanostructures. Electrochemical studies reveal that nanoCeO2 particles provide an increased electroactive surface area for loading of r-IgGs with desired orientation, resulting in enhanced electron communication between r-IgGs and electrode. BSA/r-IgGs/nano CeO2/ITO immunoelectrode exhibits improved characteristics such as linear range (0.5-6 ng dl-1), low detection limit (0.25 ng dl-1), fast response time (30 s) and high sensitivity (1.27 µA ng-1 dl-1 cm-2). The high value of the association constant (Ka, 0.9 × 1011 l mol-1) indicates the high affinity of the BSA/r-IgGs/nanoCeO2/ITO immunoelectrode to OTA.

  11. Zinc oxide nanoparticles based microfluidic immunosensor applied in congenital hypothyroidism screening.

    Science.gov (United States)

    Seia, Marco A; Pereira, Sirley V; Fernández-Baldo, Martin A; De Vito, Irma E; Raba, Julio; Messina, Germán A

    2014-07-01

    In this article, we present an innovative approach for congenital hypothyroidism (CHT) screening. This pathology is the most common preventable cause of mental retardation, affecting newborns around the world. Its consequences could be avoided with an early diagnosis through the thyrotropin (TSH) level measurement. To accomplish the determination of TSH, synthesized zinc oxide (ZnO) nanobeads (NBs) covered by chitosan (CH), ZnO-CH NBs, were covalently attached to the central channel of the designed microfluidic device. These beads were employed as platform for anti-TSH monoclonal antibody immobilization to specifically recognize and capture TSH in neonatal samples without any special pretreatment. Afterwards, the amount of this trapped hormone was quantified by horseradish peroxidase (HRP)-conjugated anti-TSH antibody. HRP reacted with its enzymatic substrate in a redox process, which resulted in the appearance of a current whose magnitude was directly proportional to the level of TSH in the neonatal sample. The structure and morphology of synthesized ZnO-CH NBs were characterized by scanning electron microscopy (SEM) and X-ray diffraction (XRD). The calculated detection limits for electrochemical detection and the enzyme-linked immunosorbent assay procedure were 0.00087 μUI mL(-1) and 0.015 μUI mL(-1), respectively, and the within- and between-assay coefficients of variation were below 6.31% for the proposed method. According to the cut-off value for TSH neonatal screening, a reasonably good limit of detection was achieved. These above-mentioned features make the system advantageous for routine clinical analysis adaptation.

  12. A Novel Method for Preparation of Gold NanoBipyramids Using Microwave Irradiation and Its Application in Immunosensors

    Science.gov (United States)

    Huynh, Trong Phat; Ngo, Vo Ke Thanh; Nguyen, Dang Giang; Nguyen, Hoang Phuong Uyen; Nghiem, Quoc Dat; Lam, Quang Vinh; Huynh, Thanh Dat

    2016-05-01

    Gold nanobipyramids (NBPs) have attracted attention for producing smart sensing devices as diagnostic tools in biotechnological and medical applications, because they show more advantageous plasmonic properties than comparable gold nanorods. Normally, NBPs were synthesized using seed-mediated growth process at room temperature. In this report, our group describes a method for synthesising of NBPs using microwave irradiation with ascorbic acid reduction and cetyltrimethylammonium bromide + silver nitrate (AgNO3) as capping agents. The advantages of this method are a highly effective approach to fast and uniform NBPs. The product was characterized by ultraviolet-visible spectroscopy, Fourier transform infrared spectroscopy, transmission electron microscopy, and x-ray powder diffraction. As an application in quartz crystal microbalance immunosensors, NBPs is conjugated with the chloramphenicol antibodies for signal amplification to detect chloramphenicol residuals in the QCM system.

  13. Amperometric glucose sensor based on nickel nanoparticles/carbon Vulcan XC-72R.

    Science.gov (United States)

    Abdel Hameed, R M

    2013-09-15

    A stable non-enzymatic glucose sensor was constructed by chemical deposition of nickel nanoparticles on carbon Vulcan XC-72R using microwave irradiation technique. The mode and time of microwave irradiation during nickel salt reduction were varied. This was found to affect the morphology of formed Ni/C powder as evidenced by TEM analysis. Nickel nanoparticles aggregation becomes more serious at longer microwave irradiation times. The electrocatalytic activity of different Ni/C samples towards glucose oxidation was studied in KOH solution by employing cyclic voltammetry and chronoamperometry techniques. Ni/C sample, prepared by pulse mode with total operating time of 150s, showed the highest oxidation current density. An excellent sensitivity value of 1349.7μAmM(-1)cm(-2) with a detection limit of 0.232μM was gained by Ni/C sensor. It also exhibits good reproducibility and long-term stability, as well as high selectivity with insignificant interference from ascorbic acid.

  14. Preparation of metal nanoband microelectrode on poly(dimethylsiloxane) for chip-based amperometric detection

    Energy Technology Data Exchange (ETDEWEB)

    Chen Shaopeng; Wu Jian; Yu Xiaodong [Key Laboratory of Analytical Chemistry for Life Science, Ministry of Education of China, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093 (China); Xu Jingjuan, E-mail: xujj@nju.edu.cn [Key Laboratory of Analytical Chemistry for Life Science, Ministry of Education of China, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093 (China); Chen Hongyuan [Key Laboratory of Analytical Chemistry for Life Science, Ministry of Education of China, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093 (China)

    2010-04-30

    We proposed herein a novel approach for fabricating nanoband microelectrodes for electrochemical detection on an electrophoresis microchip. The metal films were first obtained via region-selective electroless deposition of gold or copper films on PDMS substrates by selective region plasma oxidation through shadow masking. Both metal films show uniform surfaces with the thickness at the level of 100 nm. By casting another PDMS layer on the metal films, the cross section of the sandwich structures can be used as nanoband microelectrodes, which can be renewed just by cutting. These nanoband microelectrodes are successfully used as electrochemical detectors in microchip electrophoresis for the detection of amino acids, proteins and neurotransmitter molecules. Moreover, integrating an Au-Cu double-metal detector with a double-channel electrophoresis system, we can easily distinguish electroactive amino acids from that of non-electroactive amino acids.

  15. Amperometric sensors based on sawdust film modified electrodes: application to the electroanalysis of paraquat.

    Science.gov (United States)

    Kenne Dedzo, Gustave; Nanseu-Njiki, Charles Péguy; Ngameni, Emmanuel

    2012-09-15

    Natural or sodium hydroxide treated Ayous sawdusts were used to prepare thin film electrodes (denoted respectively as PSTFE and SSTFE). The sensors obtained exhibit good mechanical stability and a wide electrochemical potential range. Their electrochemical characterization revealed that they present a good capacity to accumulate cations, but are not useful for the electroanalysis of anions. In all cases, the signals were more intense and well defined on SSTFE compared to PSTFE. When applied to the electroanalysis of paraquat, a significant improvement of the current intensities was obtained on these electrodes compared to the bare glassy carbon electrode. The diffusion of this compound through the film which is the main process governing the electrochemical reaction at the electrode surface, is 2.2 times more important with SSTFE compared to PSTFE. After the optimization of the detection parameters, calibration curves were obtained in the concentration range 0.1-0.725 μmol L(-1) for PSTFE and 0.05-0.6 μmol L(-1) for SSTFE. The detection limits determined for a signal/noise ratio=3 are 5.49×10(-9) mol L(-1) for PSTFE and 3.02×10(-9) mol L(-1) for SSTFE.

  16. Mass Transfer in Amperometric Biosensors Based on Nanocomposite Thin Films of Redox Polymers and Oxidoreductases

    Directory of Open Access Journals (Sweden)

    Aleksandr L. Simonian

    2002-03-01

    Full Text Available Mass transfer in nanocomposite hydrogel thin films consisting of alternating layers of an organometallic redox polymer (RP and oxidoreductase enzymes was investigated. Multilayer nanostructures were fabricated on gold surfaces by the deposition of an anionic self-assembled monolayer of 11-mercaptoundecanoic acid, followed by the electrostatic binding of a cationic redox polymer, poly[vinylpyridine Os(bis-bipyridine2Clco-allylamine], and an anionic oxidoreductase. Surface plasmon resonance spectroscopy, Fourier transform infrared external reflection spectroscopy (FTIR-ERS, ellipsometry and electrochemistry were employed to characterize the assembly of these nanocomposite films. Simultaneous SPR/electrochemistry enabled real time observation of the assembly of sensing components, changes in film structure with electrode potential, and the immediate, in situ electrochemical verification of substrate-dependent current upon the addition of enzyme to the multilayer structure. SPR and FTIR-ERS studies also showed no desorption of polymer or enzyme from the nanocomposite structure when stored in aqueous environment occurred over the period of three weeks, suggesting that decreasing in substrate sensitivity were due to loss of enzymatic activity rather than loss of film compounds from the nanostructure.

  17. An Ultrasensitive Electrochemical Immunosensor for Alpha-Fetoprotein Using an Envision Complex-Antibody Copolymer as a Sensitive Label

    Directory of Open Access Journals (Sweden)

    Lei Zheng

    2012-12-01

    Full Text Available A novel strategy is presented for sensitive detection of alfa-fetoprotein (AFP, using a horseradish peroxidase (HRP-functionalized Envision antibody complex (EVC as the label. The Envision-AFP signal antibody copolymer (EVC-AFP Ab2 was composed of a dextran amine skeleton anchoring more than 100 molecules of HRP and 15 molecules of secondary antibody, and acted as a signal tag in the immunosensor. The sensor was constructed using the following steps: First, gold electrode (GE was modified with nano-gold (AuNPs by electro-deposition in HAuCl4 solution. The high affinity of the AuNPs surface facilitates direct formation of a self-assembled thiolated protein G layer. Next, the coated GE was incubated in a solution of AFP capture antibody (AFP Ab1; these antibodies attach to the thiolated protein G layer through their non-antigenic regions, leaving the antigen binding sites for binding of target analyte. Following a sandwich immunoreaction, an EVC-AFP Ab2-AFP-AFP Ab1 immunocomplex was formed on the electrode surface, allowing large amounts of HRP on the complex to produce an amplified electrocatalytic current of hydroquinone (HQ in the presence of hydrogen peroxide (H2O2. Highly amplified detection was achieved, with a detection limit of 2 pg/mL and a linear range of 0.005–0.2 ng/mL for AFP in 10 μL undiluted serum; this is near or below the normal levels of most cancer biomarker proteins in human serum. Measurements of AFP in the serum of cancer patients correlated strongly with standard enzyme-linked immunosorbent assays. These easily fabricated EVC-modified immunosensors show excellent promise for future fabrication of bioelectronic arrays. By varying the target biomolecules, this technique may be easily extended for use with other immunoassays, and thus represents a versatile design route.

  18. Determination of amitrole and urazole in water samples by capillary zone electrophoresis using simultaneous UV and amperometrical detection.

    Science.gov (United States)

    Chicharro, M; Moreno, M; Bermejo, E; Ongay, S; Zapardiel, A

    2005-12-16

    In this paper, capillary zone electrophoresis with amperometric detection (CZE-AD) was first applied to the simultaneous separation and determination of amitrole and urazole in water samples. A simple end-column electrochemical detector was used in combination with a commercially available capillary electrophoresis instrument with UV detection. The effects of several important factors were investigated to find optimum conditions. A carbon disk electrode was used as working electrode. Separation and determination of these compounds in water samples were performed in 0.030 mol l(-1) acetate buffers at pH 4.5, 25 kV as separation voltage and the samples were introduced by hydrodynamic mode for 1.5 s. Most of the studies realized showed that the direct electrochemical detection is more sensitive and selective than UV detection. Under the optimum conditions, excellent linearity was observed between peak amperometric signal and analyte concentrations in the range of 0.19-1.35 mg l(-1) for amitrole and 0.20-1.62 mg l(-1) for urazole. The detection limits were 63 and 68 microg l(-1) for amitrole and urazole, respectively. The utility of this method was demonstrated by monitoring water samples, and the assay results were satisfactory. The detection limits using a previous preconcentration step for amitrole and urazole in spiked mineral water samples were 0.6 and 1.0 microg l(-1) for amitrole and urazole, respectively.

  19. Highly selective amperometric sensor for the trace level detection of hydrazine at bismuth nanoparticles decorated graphene nanosheets modified electrode.

    Science.gov (United States)

    Devasenathipathy, Rajkumar; Mani, Veerappan; Chen, Shen-Ming

    2014-06-01

    A highly selective amperometric sensor was developed for the trace level determination of hydrazine at bismuth nanoparticles (Bi) decorated graphene nanosheets (GR) composite film modified glassy carbon electrode (GCE). GR-Bi nanocomposite has been successfully prepared via simple and facile chemical reduction approach and its structure was characterized by various techniques. Surface morphological and X-ray diffraction studies revealed the formation and high loading of Bi nanoparticles on graphene sheets. GR-Bi nanocomposite modified GCE exhibited greatly enhanced electrocatalytic performance towards electro-oxidation of hydrazine in terms of decrease in overpotential and increase in oxidation peak current (Ip). The kinetic parameters such as electron transfer coefficient (α) and diffusion coefficient (Do) of the hydrazine oxidation were determined to be 0.70 and 2.65×10(-5) cm(2) s(-1), respectively. An amperometric sensor has been fabricated which detects trace level concentration of hydrazine. The sensor exhibited a wide linear range from 20 nM to 0.28 mM and a very low detection limit (LOD) of 5 nM. Remarkably, this is the lowest LOD achieved for the determination of hydrazine in neutral pH among other reported electrochemical hydrazine sensors. In addition, the sensor selectively detects hydrazine even in the presence of 1000 fold excess quantity of common interferrants. The practical feasibility of the sensor has been assessed in water and urine samples with good recoveries. Furthermore, the sensor exhibited appreciable stability, repeatability and reproducibility results.

  20. Agent orange herbicides, organophosphate and triazinic pesticides analysis in olive oil and industrial oil mill waste effluents using new organic phase immunosensors.

    Science.gov (United States)

    Martini, Elisabetta; Merola, Giovanni; Tomassetti, Mauro; Campanella, Luigi

    2015-02-15

    New immunosensors working in organic solvent mixtures (OPIEs) for the analysis of traces of different pesticides (triazinic, organophosphates and chlorurates) present in hydrophobic matrices such as olive oil were developed and tested. A Clark electrode was used as transducer and peroxidase enzyme as marker. The competitive process took place in a chloroform-hexane 50% (V/V) mixture, while the subsequent enzymatic final measurement was performed in decane and using tert-butylhydroperoxide as substrate of the enzymatic reaction. A linear response of between about 10nM and 5.0μM was usually obtained in the presence of olive oil. Recovery tests were carried out in commercial or artisanal extra virgin olive oil. Traces of pesticides were also checked in the oily matrix, in pomace and mill wastewaters from an industrial oil mill. Immunosensors show good selectivity and satisfactory precision and recovery tests performed in olive oil gave excellent results.

  1. Anodic Oxidation and Amperometric Sensing of Hydrazine at a Glassy Carbon Electrode Modified with Cobalt (II Phthalocyanine–cobalt (II Tetraphenylporphyrin (CoPc- (CoTPP4 Supramolecular Complex

    Directory of Open Access Journals (Sweden)

    Kenneth I. Ozoemena

    2006-08-01

    Full Text Available This paper describes the electrocatalytic behaviour of a glassy carbon electrode (GCEmodified with cobalt(IIphthalocyanine (CoPc complex peripherally tetrasubstituted withcobalt(IItetraphenylporphyrin (CoTPP complexes via ether linkages (i.e., CoPc-(CoTPP4. Thefeatures of the immobilised pentamer were interrogated with cyclic voltammetry andelectrochemical impedance spectroscopy (EIS using [Fe(CN6]3-/4- as redox probe revealedenhanced electron transfer properties with kapp ≈ 18 x 10-6 cms-1 compared to that of the bareGCE (4.7 x 10-6 cms-1. The viability of this supramolecular complex as a redox mediator for theanodic oxidation and sensitive amperometric determination of hydrazine in alkaline conditions isdescribed. The electrocatalytic oxidation of hydrazine by GCE-CoPc-(CoTPP4 was characterisedwith satisfactory catalytic current response with low non-Faradaic current (ca. 30 times lowerthan the bare GCE and at much lower oxidation potential (ca. 300 mV lower than the bareGCE. A mechanism for the studied electrocatalytic reaction was proposed based on thespectrophotometric evidence that revealed the major involvement of the Co(III/Co(II redox coupleof the central CoPc species rather than the CoTPP component of the pentamer. Rate constant forthe anodic oxidation of hydrazine was estimated from chronoamperometry as ~ 3x103 M-1s-1. Theproposed amperometric sensor displayed excellent charateristics towards the determination ofhydrazine in 0.2 M NaOH ; such as low overpotentials ( 100 mV vs Ag|AgCl, very fastamperometric response time (1 s, linear concentration range of up to 230 μM, with micromolardetection limit, high sensitivity and stability.

  2. Label-free porous silicon immunosensor for broad detection of opiates in a blind clinical study and results comparison to commercial analytical chemistry techniques.

    Science.gov (United States)

    Bonanno, Lisa M; Kwong, Tai C; DeLouise, Lisa A

    2010-12-01

    In this work, we evaluate for the first time the performance of a label-free porous silicon (PSi) immunosensor assay in a blind clinical study designed to screen authentic patient urine specimens for a broad range of opiates. The PSi opiate immunosensor achieved 96% concordance with liquid chromatography-mass spectrometry/tandem mass spectrometry (LC-MS/MS) results on samples that underwent standard opiate testing (n = 50). In addition, successful detection of a commonly abused opiate, oxycodone, resulted in 100% qualitative agreement between the PSi opiate sensor and LC-MS/MS. In contrast, a commercial broad opiate immunoassay technique (CEDIA) achieved 65% qualitative concordance with LC-MS/MS. Evaluation of important performance attributes including precision, accuracy, and recovery was completed on blank urine specimens spiked with test analytes. Variability of morphine detection as a model opiate target was <9% both within-run and between-day at and above the cutoff limit of 300 ng mL(-1). This study validates the analytical screening capability of label-free PSi opiate immunosensors in authentic patient samples and is the first semiquantitative demonstration of the technology's successful clinical use. These results motivate future development of label-free PSi technology to reduce complexity and cost of diagnostic testing particularly in a point-of-care setting.

  3. Amperometric cell for subcutaneous detection of hydrogen sulfide in anesthetized experimental animals.

    Science.gov (United States)

    Nagy, L; Filotás, D; Boros, M; Pozsgai, G; Pintér, E; Nagy, G

    2014-12-01

    Hydrogen sulfide (H2S) is a toxic gas. It has been recognized that H2S evolving in biochemical reactions in living organisms has an important role in different physiologic processes. Nowadays, H2S is known as an endogenous messenger molecule. Natural sulfurous spring water has been proved beneficial in the therapy of diseases of the skin and other organs (Boros et al 2013). In vivo real-time detection of local H2S concentration is an important but challenging task.We developed a two-electrode amperometric cell for selective subcutaneous detection of H2S in anesthetized mice. The cell is a small size implantable gas sensor containing a platinum disc anode and a silver cathode. The selectivity is provided by a membrane permeable only by gases. There is a buffered reversible electrochemical mediator solution in an oxidized form inside the cell. As gaseous H2S penetrates into the cell the mediator is reduced, and +0.4 V versus the reference is employed on the platinum working electrode. The reduced mediator is oxidized on the anode surface. The current provides an analytical signal representing the concentration of H2S.Appropriate shape, size and membrane material were selected, and optimal working parameters--such as mediator concentration, pH and cell voltage--were determined in vitro. The lower limit of detection in the stirred sample solution at pH = 5.5 was as small as 9.4 × 10(-7) M and a dynamic concentration range of 0-6 × 10(-4) M could be achieved.The detecting surfaces of the cell were covered with freshly dissected mouse skin to test dermal H2S permeability. In other experiments, the cell was implanted subcutaneously in an anesthetized mouse and the animal was submerged in a buffer solution containing different concentrations of H2S so that the skin surface over the sensor was covered by the solution. Measurements of subcutaneous H2S concentration were taken. The experiments clearly proved that H2S diffuses through the skin of the live mouse.

  4. Electrochemical immunosensors for the detection of tumor markers%电化学免疫传感器在肿瘤标志物检测中的应用

    Institute of Scientific and Technical Information of China (English)

    张浩春; 吕佳; 张冰; 高文超; 李兴; 常宏宏; 魏文珑

    2016-01-01

    Tumor is one of the severe threats to human health. The death rate of malignant can mainly reduced through early diagnosis and treatment. Therefore tumor markers are of significant clinic value in the early diagnosis. With the rapid development of nanotechnology,electrochemical sensor based on nanomatericals can make the detection of tumor markers with high sensitivity and selectivity. The protocol focused on the construction principle of electrochemical immunosensors using new nanomaterials such as carbon nanomaterials,noble metal nanoparticles,oxide nanomaterials,and quantum dot nanomaterials. It also focused on the applications of those immunosensors in the detection of alpha-fetoprotein,prostate antigen,carcinoembryonic antigen,and other tumor markers. The advantages and disadvantages of electrochemistrical sensors constructed on different nanomaterials in the detection of various tumor markers are analyzed and summarized. It is concluded that future development of the electrochemical immunosensors should be focus on miniaturization,high capacity, and commercialization of fast repoense,on-line,and real-time detection of tumor markers.%肿瘤是严重威胁人类健康的疾病之一,降低恶性肿瘤死亡率的主要途径是早期诊断和治疗,肿瘤标志物在肿瘤早期诊断中具有重要的临床应用价值。随着纳米技术的迅猛发展,基于纳米材料构建的电化学传感器可实现对肿瘤标志物的检测,且具有检测灵敏度高、选择性好等优点。本文重点综述了碳纳米材料、贵金属纳米材料、氧化物纳米材料、量子点纳米材料等新型纳米材料电化学免疫传感器的构建原理及其在甲胎蛋白、前列腺抗原、癌胚抗原等肿瘤标志物检测中的应用,分析总结了基于不同纳米材料构建的电化学传感器在各种肿瘤标志物检测中的优缺点,并展望了电化学传感器的发展趋势,提出未来电化学免疫传感器应以微型

  5. A novel amperometric alcohol biosensor developed in a 3rd generation bioelectrode platform using peroxidase coupled ferrocene activated alcohol oxidase as biorecognition system.

    Science.gov (United States)

    Chinnadayyala, Somasekhar R; Kakoti, Ankana; Santhosh, Mallesh; Goswami, Pranab

    2014-05-15

    Alcohol oxidase (AOx) with a two-fold increase in efficiency (Kcat/Km) was achieved by physical entrapment of the activator ferrocene in the protein matrix through a simple microwave based partial unfolding technique and was used to develop a 3rd generation biosensor for improved detection of alcohol in liquid samples. The ferrocene molecules were stably entrapped in the AOx protein matrix in a molar ratio of ~3:1 through electrostatic interaction with the Trp residues involved in the functional activity of the enzyme as demonstrated by advanced analytical techniques. The sensor was fabricated by immobilizing ferrocene entrapped alcohol oxidase (FcAOx) and sol-gel chitosan film coated horseradish peroxidase (HRP) on a multi-walled carbon nanotube (MWCNT) modified glassy carbon electrode through layer-by-layer technique. The bioelectrode reactions involved the formation of H2O2 by FcAOx biocatalysis of substrate alcohol followed by HRP-catalyzed reduction of the liberated H2O2 through MWCNT supported direct electron transfer mechanism. The amperometric biosensor exhibited a linear response to alcohol in the range of 5.0 × 10(-6) to 30 × 10(-4)mol L(-1) with a detection limit of 2.3 × 10(-6) mol L(-1), and a sensitivity of 150 µA mM(-1) cm(-2). The biosensor response was steady for 28 successive measurements completed in a period of 5h and retained ~90% of the original response even after four weeks when stored at 4 °C. The biosensor was successfully applied for the determination of alcohol in commercial samples and its performance was validated by comparing with the data obtained by GC analyses of the samples.

  6. Development of sensitive amperometric hydrogen peroxide sensor using a CuNPs/MB/MWCNT-C{sub 60}-Cs-IL nanocomposite modified glassy carbon electrode

    Energy Technology Data Exchange (ETDEWEB)

    Roushani, Mahmoud, E-mail: mahmoudroushani@yahoo.com; Bakyas, Kobra; Zare Dizajdizi, Behruz

    2016-07-01

    A sensitive hydrogen peroxide (H{sub 2}O{sub 2}) sensor was constructed based on copper nanoparticles/methylene blue/multiwall carbon nanotubes–fullerene–chitosan–ionic liquid (CuNPs/MB/MWCNTs–C{sub 60}–Cs–IL) nanocomposites. The MB/MWCNTs–C{sub 60}–Cs–IL and CuNPs were modified glassy carbon electrode (GCE) by the physical adsorption and electrodeposition of copper nitrate solution, respectively. The physical morphology and chemical composition of the surface of modified electrode was investigated by scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS), respectively. The electrochemical properties of CuNPs/MB/MWCNTs–C{sub 60}–Cs–IL/GCE were investigated by cyclic voltammetry (CV) and amperometry techniques and the sensor exhibited remarkably strong electrocatalytic activities toward the reduction of hydrogen peroxide. The peak currents possess a linear relationship with the concentration of H{sub 2}O{sub 2} in the range of 0.2 μM to 2.0 mM, and the detection limit is 55.0 nM (S/N = 3). In addition, the modified electrode was used to determine H{sub 2}O{sub 2} concentration in human blood serum sample with satisfactory results. - Highlights: • CuNPs/MB/MWCNT-C{sub 60}-Cs-IL/GC electrode was constructed by layer-by-layer method. • The catalytic performance