Magnetic separation of antibiotics by electrochemical magnetic seeding

International Nuclear Information System (INIS)

Ihara, I; Toyoda, K; Beneragama, N; Umetsu, K

2009-01-01

Magnetic separation of several classes of antibiotics was investigated using electrochemical magnetic seeding. Electrocoagulation with a sacrificial anode followed by addition of magnetite particles was applied for the magnetic seeding of antibiotics. With electrochemical magnetic seeding using an iron anode, tetracycline antibiotics (oxytetracycline, chlortetracycline, doxycycline and tetracycline) and cephalosporin antibiotic (cefdinir) were rapidly removed from synthetic wastewater by magnetic separation using a neodymium magnet. Iron and aluminium anodes were suitable for magnetic seeding of the antibiotics. The results indicated that the ability of antibiotics to form strong complex with iron and aluminium allowed the higher removal by magnetic separation. This method would be appropriate for rapid treatment of antibiotics in wastewater.

Nanobody-based electrochemical immunoassay for Bacillus thuringiensis Cry1Ab toxin by detecting the enzymatic formation of polyaniline

International Nuclear Information System (INIS)

Zhu, Min; Li, Guanghui; Li, Min; Zhou, Zikai; Liu, Hong; Lei, Hongtao; Shen, Yanfei; Wan, Yakun

2015-01-01

We describe an electrochemical immunoassay for the Cry1Ab toxin that is produced by Bacillus thuringiensis. It is making use of a nanobody (a heavy-chain only antibody) that was selected from an immune phage displayed library. A biotinylated primary nanobody and a HRP-conjugated secondary nanobody were applied in a sandwich immunoassay where horseradish peroxidase (HRP) is used to produce polyaniline (PANI) from aniline. PANI can be easily detected by differential pulse voltammetry at a working voltage as low as 40 mV (vs. Ag/AgCl) which makes the assay fairly selective. This immunoassay for Cry1Ab has an analytical range from 0.1 to 1000 ng∙mL -1 and a 0.07 ng∙mL -1 lower limit of detection. The average recoveries of the toxin from spiked samples are in the range from 102 to 114 %, with a relative standard deviation of <7.5 %. The results demonstrated that the assay represented an attractive alternative to existing immunoassays in enabling affordable, sensitive, robust and specific determination of this toxin. (author)

Chemiluminescence enzyme immunoassay based on magnetic nanoparticles for detection of hepatocellular carcinoma marker glypican-3

Directory of Open Access Journals (Sweden)

Qian-Yun Zhang

2011-08-01

Full Text Available Glypican-3 (GPC3 is reported as a great promising tumor marker for hepatocellular carcinoma (HCC diagnosis. Highly sensitive and accurate analysis of serum GPC3 (sGPC3, in combination with or instead of traditional HCC marker alpha-fetoprotein (AFP, is essential for early diagnosis of HCC. Biomaterial-functionalized magnetic particles have been utilized as solid supports with good biological compatibility for sensitive immunoassay. Here, the magnetic nanoparticles (MnPs and magnetic microparticles (MmPs with carboxyl groups were further modified with streptavidin, and applied for the development of chemiluminescence enzyme immunoassay (CLEIA. After comparing between MnPs- and MmPs-based CLEIA, MnPs-based CLEIA was proved to be a better method with less assay time, greater sensitivity, better linearity and longer chemiluminescence platform. MnPs-based CLEIA was applied for detection of sGPC3 in normal liver, hepatocirrhosis, secondary liver cancer and HCC serum samples. The results indicated that sGPC3 was effective in diagnosis of HCC with high performance. Keywords: Magnetic nanoparticle, Magnetic microparticle, Chemiluminescence enzyme immunoassay, Glypican-3, Hepatocellular carcinoma

Sensitive and rapid immunoassay for parathyroid hormone using magnetic particle labels and magnetic actuation

NARCIS (Netherlands)

Dittmer, W.U.; Kievit, de P.; Prins, M.W.J.; Vissers, J.L.M.; Mersch, M.E.C.; Martens, M.F.W.C.

2008-01-01

A rapid method for the sensitive detection of proteins using actuated magnetic particle labels, which are measured with a giant magneto-resistive (GMR) biosensor, is described. The technique involves a 1-step sandwich immunoassay with no fluid replacement steps. The various assay binding reactions

Electrochemical Immunoassay Using Open Circuit Potential Detection Labeled by Platinum Nanoparticles

Directory of Open Access Journals (Sweden)

Kanokwan Charoenkitamorn

2018-02-01

Full Text Available In this work, a simple electrochemical immunoassay based on platinum nanoparticles (PtNPs using open circuit potential (OCP detection was developed. The detection of human chorionic gonadotropin hormone (hCG as a model analyte, was demonstrated by direct electrical detection of PtNPs in hydrazine solution using OCP measurement without any application of either potential or current to the system. Disposable screen-printed carbon electrodes (SPCEs were utilized for the development of our immunosensor, which required a sample volume as small as 2 μL. After preparation of a sandwich-type immunosystem, hydrazine solution was dropped on the electrode’s surface, which was followed immediately by electrical detection using OCP. The change of the OCP signal originated from electrocatalytic oxidation of the hydrazine on PtNPs. Under the optimal conditions of a pH of 6.0 and a hydrazine concentration of 1 mM, a detection limit of 0.28 ng mL−1 and a linearity of 0–10 ng mL−1 were obtained. The PtNP-based OCP method is a simpler electrochemical detection procedure than those obtained from other electrochemical methods and has an acceptable sensitivity and reproducibility. The simplicity of the detection procedure and the cost-effectiveness of the disposable SPCE illustrate the attractive benefits of this sensor. Moreover, it could be applied to a simplified and miniaturized diagnostic system with minimal user manipulation.

Magnetic Particle-Based Immunoassay of Phosphorylated p53 Using Protein-Cage Templated Lead Phosphate and Carbon Nanospheres for Signal Amplification

Energy Technology Data Exchange (ETDEWEB)

Chen, Aiqiong; Bao, Yuanwu; Ge, Xiaoxiao; Shin, Yongsoon; Du, Dan; Lin, Yuehe

2012-11-20

Phosphorylated p53 at serin 15 (phospho-p53-15) is a potential biomarker of Gamma-radiation exposure. In this paper, we described a new magnetic particles (MPs)-based electrochemical immunoassay of human phospho-p53-15 using carbon nanospheres (CNS) and protein-cage templated lead phosphate nanoparticles for signal amplification. Greatly enhanced sensitivity was achieved by three aspects: 1) The protein-cage nanoparticle (PCN) and p53-15 signal antibody (p53-15 Ab2) are linked to CNS (PCNof each apoferritin; 3) MPs capture a large amount of primary antibodies. Using apoferritin templated metallic phosphate instead of enzyme as label has the advantage of eliminating the addition of mediator or immunoreagents and thus makes the immunoassay system simpler. The subsequent stripping voltammetric analysis of the released lead ions were detected on a disposable screen printed electrode. The response current was proportional to the phospho-p53-15 concentration in the range of 0.02 to 20 ng mL-1 with detection limit of 0.01 ng mL-1. This method shows a good stability, reproducibility and recovery.

Sequential injection chemiluminescence immunoassay for nonionic surfactants by using magnetic microbeads

International Nuclear Information System (INIS)

Zhang Ruiq; Nakajima, Hizuru; Soh, Nobuaki; Nakano, Koji; Masadome, Takashi; Nagata, Kazumi; Sakamoto, Kazuhira; Imato, Toshihiko

2007-01-01

A rapid and sensitive immunoassay based on a sequential injection analysis (SIA) using magnetic microbeads for the determination of alkylphenol polyethoxylates (APnEOs) is described. An SIA system was constructed from a syringe pump, a switching valve, a flow-through type immunoreaction cell equipped with a photon counting unit and a neodymium magnet. Magnetic beads, to which an anti-APnEOs monoclonal antibody was immobilized, were used as a solid support in an immunoassay. The introduction, trapping and release of the magnetic beads in and from the immunoreaction cell were controlled by means of a neodymium magnet and adjusting the flow of a carrier solution. The immunoassay was based on an indirect competitive immunoreaction of an anti-APnEOs monoclonal antibody immobilized on the magnetic beads with a sample APnEOs and a horseradish peroxidase (HRP)-labeled APnEOs in the same sample solution, and was based on the subsequent chemiluminscence reaction of HRP on the magnetic microbeads with a luminol solution containing hydrogen peroxide and p-iodophenol. The anti-APnEOs antibody was immobilized on the magnetic microbeads by coupling the antibody with the magnetic beads after activation of a carboxylate moiety on the surface of the magnetic beads that had been coated with a polylactic acid film. The antibody immobilized magnetic beads were introduced in the immunoreaction cell and trapped in it by the neodymium magnet, which was equipped beneath the immunoreaction cell. An APnEOs sample solution containing the HRP-labeled APnEOs at a constant concentration, and a luminol solution containing hydrogen peroxide and p-iodophenol were sequentially introduced into the immunoreaction cell, according to an SIA programmed sequence. Chemiluminescence emission was monitored by means of a photon counting unit located at the upper side of the immunoreaction cell by collecting the emitted light with a lens. A typical sigmoidal calibration curve was obtained, when the logarithm

An ultrasensitive chemiluminescence immunoassay of chloramphenicol based on gold nanoparticles and magnetic beads.

Science.gov (United States)

Tao, Xiaoqi; Jiang, Haiyang; Yu, Xuezhi; Zhu, Jinghui; Wang, Xia; Wang, Zhanhui; Niu, Lanlan; Wu, Xiaoping; Shen, Jianzhong

2013-05-01

A competitive, direct, chemiluminescent immunoassay based on a magnetic beads (MBs) separation and gold nanoparticles (AuNPs) labelling technique to detect chloramphenicol (CAP) has been developed. Horseradish peroxidase (HRP)-labelled anti-CAP monoclonal antibody conjugated with AuNPs and antigen-immobilized MBs were prepared. After optimization parameters of immunocomplex MBs, the IC50 values of chemiluminescence magnetic nanoparticles immunoassay (CL-MBs-nano-immunoassay) were 0.017 µg L(-1) for extract method I and 0.17 µg L(-1) for extract method II. The immunoassay with two extract methods was applied to detect CAP in milk. Comparison of these two extract methods showed that extract method I was advantageous in better sensitivity, in which the sensitivity was 10 times compared to that of extract method II, while extract method II was superior in simple operation, suitable for high throughout screen. The recoveries were 86.7-98.0% (extract method I) and 80.0-103.0% (extract method II), and the coefficients of variation (CVs) were all recovery with both extract methods and high correlation with traditional ELISA kit in milk system confirmed that the immunomagnetic assay based on AuNPs exhibited promising potential in rapid field screening for trace CAP analysis. Copyright © 2013 John Wiley & Sons, Ltd.

Proximity hybridization-regulated catalytic DNA hairpin assembly for electrochemical immunoassay based on in situ DNA template-synthesized Pd nanoparticles

International Nuclear Information System (INIS)

Zhou, Fuyi; Yao, Yao; Luo, Jianjun; Zhang, Xing; Zhang, Yu; Yin, Dengyang; Gao, Fenglei; Wang, Po

2017-01-01

Novel hybridization proximity-regulated catalytic DNA hairpin assembly strategy has been proposed for electrochemical immunoassay based on in situ DNA template-synthesized Pd nanoparticles as signal label. The DNA template-synthesized Pd nanoparticles were characterized with atomic force microscopic and X-ray photoelectron spectroscopy. The highly efficient electrocatalysis by DNA template synthesized Pd nanoparticles for NaBH 4 oxidation produced an intense detection signal. The label-free electrochemical method achieved the detection of carcinoembryonic antigen (CEA) with a linear range from 10 −15 to 10 −11  g mL −1 and a detection limit of 0.43 × 10 −15  g mL −1 . Through introducing a supersandwich reaction to increase the DNA length, the electrochemical signal was further amplified, leading to a detection limit of 0.52 × 10 −16  g mL −1 . And it rendered satisfactory analytical performance for the determination of CEA in serum samples. Furthermore, it exhibited good reproducibility and stability; meanwhile, it also showed excellent specificity due to the specific recognition of antigen by antibody. Therefore, the DNA template synthesized Pd nanoparticles based signal amplification approach has great potential in clinical applications and is also suitable for quantification of biomarkers at ultralow level. - Graphical abstract: A novel label-free and enzyme-free electrochemical immunoassay based on proximity hybridization-regulated catalytic DNA hairpin assemblies for recycling of the CEA. - Highlights: • A novel enzyme-free electrochemical immunosensor was developed for detection of CEA. • The signal amplification was based on catalytic DNA hairpin assembly and DNA-template-synthesized Pd nanoparticles. • The biosensor could detect CEA down to 0.52 × 10 −16  g mL −1 level with a dynamic range spanning 5 orders of magnitude.

Ferrocenyl-doped silica nanoparticles as an immobilized affinity support for electrochemical immunoassay of cancer antigen 15-3

International Nuclear Information System (INIS)

Hong Chenglin; Yuan Ruo; Chai Yaqin; Zhuo Ying

2009-01-01

The aim of this study is to elaborate a simple and sensitive electrochemical immunoassay using ferrocenecarboxylic (Fc-COOH)-doped silica nanoparticles (SNPs) as an immobilized affinity support for cancer antigen 15-3 (CA 15-3) detection. The Fc-COOH-doped SNPs with redox-active were prepared by using a water-in-oil microemulsion method. The use of colloidal silica could prevent the leakage of Fc-COOH and were easily modified with trialkoxysilane reagents for covalent conjugation of CA 15-3 antibodies (anti-CA 15-3). The Fc-COOH-doped SNPs were characterized by X-ray photoelectron spectroscopy (XPS) and transmission electron microscopy (TEM). The fabrication process of the electrochemical immunosensor was demonstrated by using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) techniques. Under optimal conditions, the developed immunosensor showed good linearity at the studied concentration range of 2.0-240 U mL -1 with a coefficient 0.9986 and a detection limit of 0.64 U mL -1 at S/N = 3

Ferrocenyl-doped silica nanoparticles as an immobilized affinity support for electrochemical immunoassay of cancer antigen 15-3.

Science.gov (United States)

Hong, Chenglin; Yuan, Ruo; Chai, Yaqin; Zhuo, Ying

2009-02-09

The aim of this study is to elaborate a simple and sensitive electrochemical immunoassay using ferrocenecarboxylic (Fc-COOH)-doped silica nanoparticles (SNPs) as an immobilized affinity support for cancer antigen 15-3 (CA 15-3) detection. The Fc-COOH-doped SNPs with redox-active were prepared by using a water-in-oil microemulsion method. The use of colloidal silica could prevent the leakage of Fc-COOH and were easily modified with trialkoxysilane reagents for covalent conjugation of CA 15-3 antibodies (anti-CA 15-3). The Fc-COOH-doped SNPs were characterized by X-ray photoelectron spectroscopy (XPS) and transmission electron microscopy (TEM). The fabrication process of the electrochemical immunosensor was demonstrated by using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) techniques. Under optimal conditions, the developed immunosensor showed good linearity at the studied concentration range of 2.0-240 UmL(-1) with a coefficient 0.9986 and a detection limit of 0.64 UmL(-1) at S/N=3.

Magnetofluorescent nanocomposites and quantum dots used for optimal application in magnetic fluorescence-linked immunoassay.

Science.gov (United States)

Tsai, H Y; Li, S Y; Fuh, C Bor

2018-03-01

Magnetofluorescent nanocomposites with optimal magnetic and fluorescent properties were prepared and characterized by combining magnetic nanoparticles (iron oxide@polymethyl methacrylate) with fluorescent nanoparticles (rhodamine 6G@mSiO 2 ). Experimental parameters were optimized to produce nanocomposites with high magnetic susceptibility and fluorescence intensity. The detection of a model biomarker (alpha-fetoprotein) was used to demonstrate the feasibility of applying the magnetofluorescent nanocomposites combined with quantum dots and using magnetic fluorescence-linked immunoassay. The magnetofluorescent nanocomposites enable efficient mixing, fast re-concentration, and nanoparticle quantization for optimal reactions. Biofunctional quantum dots were used to confirm the alpha-fetoprotein (AFP) content in sandwich immunoassay after mixing and washing. The analysis time was only one third that required in ELISA. The detection limit was 0.2 pg mL -1 , and the linear range was 0.68 pg mL -1 -6.8 ng mL -1 . This detection limit is lower, and the linear range is wider than those of ELISA and other methods. The measurements made using the proposed method differed by less than 13% from those obtained using ELISA for four AFP concentrations (0.03, 0.15, 0.75, and 3.75 ng mL -1 ). The proposed method has a considerable potential for biomarker detection in various analytical and biomedical applications. Graphical abstract Magnetofluorescent nanocomposites combined with fluorescent quantum dots were used in magnetic fluorescence-linked immunoassay.

Proximity hybridization-regulated catalytic DNA hairpin assembly for electrochemical immunoassay based on in situ DNA template-synthesized Pd nanoparticles

Energy Technology Data Exchange (ETDEWEB)

Zhou, Fuyi [School of Chemistry and Chemical Engineering, Jiangsu Normal University, Xuzhou 221116 (China); Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Department of Pharmaceutical Analysis, School of Pharmacy, Xuzhou Medical College, 221004, Xuzhou (China); Yao, Yao; Luo, Jianjun; Zhang, Xing; Zhang, Yu; Yin, Dengyang [Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Department of Pharmaceutical Analysis, School of Pharmacy, Xuzhou Medical College, 221004, Xuzhou (China); Gao, Fenglei, E-mail: jsxzgfl@sina.com [Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Department of Pharmaceutical Analysis, School of Pharmacy, Xuzhou Medical College, 221004, Xuzhou (China); Wang, Po, E-mail: wangpo@jsnu.edu.cn [School of Chemistry and Chemical Engineering, Jiangsu Normal University, Xuzhou 221116 (China)

2017-05-29

Novel hybridization proximity-regulated catalytic DNA hairpin assembly strategy has been proposed for electrochemical immunoassay based on in situ DNA template-synthesized Pd nanoparticles as signal label. The DNA template-synthesized Pd nanoparticles were characterized with atomic force microscopic and X-ray photoelectron spectroscopy. The highly efficient electrocatalysis by DNA template synthesized Pd nanoparticles for NaBH{sub 4} oxidation produced an intense detection signal. The label-free electrochemical method achieved the detection of carcinoembryonic antigen (CEA) with a linear range from 10{sup −15} to 10{sup −11} g mL{sup −1} and a detection limit of 0.43 × 10{sup −15} g mL{sup −1}. Through introducing a supersandwich reaction to increase the DNA length, the electrochemical signal was further amplified, leading to a detection limit of 0.52 × 10{sup −16} g mL{sup −1}. And it rendered satisfactory analytical performance for the determination of CEA in serum samples. Furthermore, it exhibited good reproducibility and stability; meanwhile, it also showed excellent specificity due to the specific recognition of antigen by antibody. Therefore, the DNA template synthesized Pd nanoparticles based signal amplification approach has great potential in clinical applications and is also suitable for quantification of biomarkers at ultralow level. - Graphical abstract: A novel label-free and enzyme-free electrochemical immunoassay based on proximity hybridization-regulated catalytic DNA hairpin assemblies for recycling of the CEA. - Highlights: • A novel enzyme-free electrochemical immunosensor was developed for detection of CEA. • The signal amplification was based on catalytic DNA hairpin assembly and DNA-template-synthesized Pd nanoparticles. • The biosensor could detect CEA down to 0.52 × 10{sup −16} g mL{sup −1} level with a dynamic range spanning 5 orders of magnitude.

The fabrication of magnetic particle-based chemiluminescence immunoassay for human epididymis protein-4 detection in ovarian cancer

Directory of Open Access Journals (Sweden)

Xiaoling Fu

2018-03-01

Full Text Available The magnetic particles have a significant influence on the immunoassay detection and cancer therapy. Herein, the chemiluminescence immunoassay combined with the magnetic particles (MPCLIA was presented for the clinical determination and analysis of human epididymis protein 4 (HE4 in the human serum. Under the optimized experiment conditions, the secure MPCLIA method can detect HE4 in the broader range of 0–1000 pmol/L, with a lower detection limit of 1.35 pmol/L. The satisfactory recovery rate of the method in the serum ranged from 83.62% to 105.10%, which was well within the requirement of clinical analysis. Moreover, the results showed the good correlation with enzyme-linked immunosorbent assay (ELISA, with the correlation coefficient of 0.9589. This proposed method has been successfully applied to the clinical determination of HE4 in the human serum. Keywords: Chemiluminescence immunoassay, Magnetic particles, Human epididymis protein 4

Redox-Magnetohydrodynamic Microfluidics Without Channels and Compatible with Electrochemical Detection Under Immunoassay Conditions

Science.gov (United States)

Weston, Melissa C.; Nash, Christena K.; Fritsch, Ingrid

2010-01-01

A unique capability of redox-magnetohydrodynamics (redox-MHD) for handling liquids on a small scale was demonstrated. A 1.2-μL solution plug was pumped from an injection site to a detector without the need for a channel to direct the flow. The redox pumping species did not interfere with enzymatic activity in a solution compatible with enzyme-linked immunoassays. Alkaline phosphatase (AP), a common enzyme label, converted p-aminophenyl phosphate (PAPP) to p-aminophenol (PAPR) in the presence of 2.5 mM Ru(NH3)6Cl2 and 2.5 mM Ru(NH3)6 Cl3, in 0.1 M Tris buffer (pH=9). A solution plug containing PAPP (no AP) was pumped through the surrounding solution containing AP (no PAPP), and the enzymatically-generated PAPR was easily detected and distinguishable electrochemically from the pumping species with square wave voltammetry down to 0.1 mM concentrations. The test device consisted of a silicon chip containing individually-addressable microband electrodes, placed on a 0.5-T NdFeB permanent magnet with the field oriented perpendicular to the chip. A 8.0-mm wide × 15.5-mm long × 1.5-mm high volume of solution was contained by a poly(dimethylsiloxane) gasket and capped with a glass slide. A steady-state fluid velocity of ~30 μm/s was generated in a reinforcing flow configuration between oppositely polarized sets of pumping electrodes with ~2.1 μA. PMID:20681513

Targeted deposition of antibodies on a multiplex CMOS microarray and optimization of a sensitive immunoassay using electrochemical detection.

Directory of Open Access Journals (Sweden)

John Cooper

2010-03-01

Full Text Available The CombiMatrix ElectraSense microarray is a highly multiplex, complementary metal oxide semiconductor with 12,544 electrodes that are individually addressable. This platform is commercially available as a custom DNA microarray; and, in this configuration, it has also been used to tether antibodies (Abs specifically on electrodes using complementary DNA sequences conjugated to the Abs.An empirical method is described for developing and optimizing immunoassays on the CombiMatrix ElectraSense microarray based upon targeted deposition of polypyrrole (Ppy and capture Ab. This process was automated using instrumentation that can selectively apply a potential or current to individual electrodes and also measure current generated at the electrodes by an enzyme-enhanced electrochemical (ECD reaction. By designating groups of electrodes on the array for different Ppy deposition conditions, we determined that the sensitivity and specificity of a sandwich immunoassay for staphylococcal enterotoxin B (SEB is influenced by the application of different voltages or currents and the application time. The sandwich immunoassay used a capture Ab adsorbed to the Ppy and a reporter Ab labeled for fluorescence detection or ECD, and results from these methods of detection were different.Using Ppy deposition conditions for optimum results, the lower limit of detection for SEB using the ECD assay was between 0.003 and 0.01 pg/ml, which represents an order of magnitude improvement over a conventional enzyme-linked immunosorbant assay. In the absence of understanding the variables and complexities that affect assay performance, this highly multiplexed electrode array provided a rapid, high throughput, and empirical approach for developing a sensitive immunoassay.

Magnetic field effects on electrochemical metal depositions

Directory of Open Access Journals (Sweden)

Andreas Bund, Adriana Ispas and Gerd Mutschke

2008-01-01

Full Text Available This paper discusses recent experimental and numerical results from the authors' labs on the effects of moderate magnetic (B fields in electrochemical reactions. The probably best understood effect of B fields during electrochemical reactions is the magnetohydrodynamic (MHD effect. In the majority of cases it manifests itself in increased mass transport rates which are a direct consequence of Lorentz forces in the bulk of the electrolyte. This enhanced mass transport can directly affect the electrocrystallization. The partial currents for the nucleation of nickel in magnetic fields were determined using an in situ micro-gravimetric technique and are discussed on the basis of the nucleation model of Heerman and Tarallo. Another focus of the paper is the numerical simulation of MHD effects on electrochemical metal depositions. A careful analysis of the governing equations shows that many MHD problems must be treated in a 3D geometry. In most cases there is a complex interplay of natural and magnetically driven convection.

Electrochemical immunoassay for thyroxine detection using cascade catalysis as signal amplified enhancer and multi-functionalized magnetic graphene sphere as signal tag

Energy Technology Data Exchange (ETDEWEB)

Han, Jing; Zhuo, Ying, E-mail: yingzhuo@swu.edu.cn; Chai, Yaqin; Yu, Yanqing; Liao, Ni; Yuan, Ruo, E-mail: yuanruo@swu.edu.cn

2013-08-06

Graphical abstract: -- Highlights: •A reusable electrochemical immunosensor is developed for thyroxine detection. •Cascade catalysis as signal amplified enhancer. •Multi-functionalized magnetic graphene sphere as signal tag. •The novel strategy has the advantages of high sensitivity, good selectivity and reproducibility. -- Abstract: This paper constructed a reusable electrochemical immunosensor for the detection of thyroxine at an ultralow concentration using cascade catalysis of cytochrome c (Cyt c) and glucose oxidase (GOx) as signal amplified enhancer. It is worth pointing out that numerous Cyt c and GOx were firstly carried onto the double-stranded DNA polymers based on hybridization chain reaction (HCR), and then the amplified responses could be achieved by cascade catalysis of Cyt c and GOx recycling with the help of glucose. Moreover, multi-functionalized magnetic graphene sphere was synthesized and used as signal tag, which not only exhibited good mechanical properties, large surface area and an excellent electron transfer rate of graphene, but also possessed excellent redox activity and desirable magnetic property. With a sandwich-type immunoreaction, the proposed cascade catalysis amplification strategy could greatly enhance the sensitivity for the detection of thyroxine. Under the optimal conditions, the immunosensor showed a wide linear ranged from 0.05 pg mL{sup −1} to 5 ng mL{sup −1} and a low detection limit down to 15 fg mL{sup −1}. Importantly, the proposed method offers promise for reproducible and cost-effective analysis of biological samples.

Electrochemical Solution Growth of Magnetic Nitrides

Energy Technology Data Exchange (ETDEWEB)

Monson, Todd C. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Pearce, Charles [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

2014-10-01

Magnetic nitrides, if manufactured in bulk form, would provide designers of transformers and inductors with a new class of better performing and affordable soft magnetic materials. According to experimental results from thin films and/or theoretical calculations, magnetic nitrides would have magnetic moments well in excess of current state of the art soft magnets. Furthermore, magnetic nitrides would have higher resistivities than current transformer core materials and therefore not require the use of laminates of inactive material to limit eddy current losses. However, almost all of the magnetic nitrides have been elusive except in difficult to reproduce thin films or as inclusions in another material. Now, through its ability to reduce atmospheric nitrogen, the electrochemical solution growth (ESG) technique can bring highly sought after (and previously inaccessible) new magnetic nitrides into existence in bulk form. This method utilizes a molten salt as a solvent to solubilize metal cations and nitrogen ions produced electrochemically and form nitrogen compounds. Unlike other growth methods, the scalable ESG process can sustain high growth rates (~mm/hr) even under reasonable operating conditions (atmospheric pressure and 500 °C). Ultimately, this translates into a high throughput, low cost, manufacturing process. The ESG process has already been used successfully to grow high quality GaN. Below, the experimental results of an exploratory express LDRD project to access the viability of the ESG technique to grow magnetic nitrides will be presented.

A Novel Colorimetric Immunoassay Utilizing the Peroxidase Mimicking Activity of Magnetic Nanoparticles

Directory of Open Access Journals (Sweden)

Hyun Gyu Park

2013-05-01

Full Text Available A simple colorimetric immunoassay system, based on the peroxidase mimicking activity of Fe3O4 magnetic nanoparticles (MNPs, has been developed to detect clinically important antigenic molecules. MNPs with ca. 10 nm in diameter were synthesized and conjugated with specific antibodies against target molecules, such as rotaviruses and breast cancer cells. Conjugation of the MNPs with antibodies (MNP-Abs enabled specific recognition of the corresponding target antigenic molecules through the generation of color signals arising from the colorimetric reaction between the selected peroxidase substrate, 3,3',5,5'-tetramethylbenzidine (TMB and H2O2. Based on the MNP-promoted colorimetric reaction, the target molecules were detected and quantified by measuring absorbance intensities corresponding to the oxidized form of TMB. Owing to the higher stabilities and economic feasibilities of MNPs as compared to horseradish peroxidase (HRP, the new colorimetric system employing MNP-Abs has the potential of serving as a potent immunoassay that should substitute for conventional HRP-based immunoassays. The strategy employed to develop the new methodology has the potential of being extended to the construction of simple diagnostic systems for a variety of biomolecules related to human cancers and infectious diseases, particularly in the realm of point-of-care applications.

Novel Electrochemical Phenomena in Magnetic Fields(Research in High Magnetic Fields)

OpenAIRE

Mogi, Iwao; Kamiko, Masao

1996-01-01

Recent two topics are given of electrochemical studies in steady magnetic fields at the High Field Laboratory of Tohoku University. One is the magnetic-field-induced diffusion-limited-aggregation in the pattern formation of silver electrodeposits . The other is the magnetic field effect on the learning effect in a dopant-exchange process of an organic conducting polymer polypyrrole.

Electrochemical synthesis of CORE-shell magnetic nanowires

KAUST Repository

Ovejero, Jesús G.

2015-04-16

(Fe, Ni, CoFe) @ Au core-shell magnetic nanowires have been synthesized by optimized two-step potentiostatic electrodeposition inside self-assembled nanopores of anodic aluminium templates. The optimal electrochemical parameters (e.g., potential) have been firstly determined for the growth of continuous Au nanotubes at the inner wall of pores. Then, a magnetic core was synthesized inside the Au shells under suitable electrochemical conditions for a wide spectrum of single elements and alloy compositions (e.g., Fe, Ni and CoFe alloys). Novel opportunities offered by such nanowires are discussed particularly the magnetic behavior of (Fe, Ni, CoFe) @ Au core-shell nanowires was tested and compared with that of bare TM nanowires. These core-shell nanowires can be released from the template so, opening novel opportunities for biofunctionalization of individual nanowires.

Ultrasensitive electrochemical immunoassay of staphylococcal enterotoxin B in food using enzyme-nanosilica-doped carbon nanotubes for signal amplification.

Science.gov (United States)

Tang, Dianping; Tang, Juan; Su, Biling; Chen, Guonan

2010-10-27

A new sandwich-type electrochemical immunoassay for ultrasensitive detection of staphylococcal enterotoxin B (SEB) in food was developed using horseradish peroxidase-nanosilica-doped multiwalled carbon nanotubes (HRPSiCNTs) for signal amplification. Rabbit polyclonal anti-SEB antibodies immobilized on the screen-printed carbon electrode (SPCE) and covalently bound to the HRPSiCNTs were used as capture antibodies and detection antibodies, respectively. In the presence of SEB analyte, the sandwich-type immunocomplex could be formed between the immobilized anti-SEB on the SPCE and anti-SEB-labeled HRPSiCNTs, and the carried HRP could catalyze the electrochemical reduction of H2O2 with the help of thionine. The high content of HRP in the HRPSiCNTs could greatly amplify the electrochemical signal. Under optimal conditions, the reduction current increased with the increase of SEB in the sample, and exhibited a dynamic range of 0.05-15 ng/mL with a low detection limit (LOD) of 10 pg/mL SEB (at 3σ). Intra- and interassay coefficients of variation were below 10%. In addition, the assay was evaluated with SEB spiked samples including watermelon juice, soymilk, apple juice, and pork food, receiving excellent correlation with results from commercially available enzyme-linked immunosorbent assay (ELISA).

Electrochemical analysis of gold-coated magnetic nanoparticles for detecting immunological interaction

International Nuclear Information System (INIS)

Pham, Thao Thi-Hien; Sim, Sang Jun

2010-01-01

An electrochemical impedance immunosensor was developed for detecting the immunological interaction between human immunoglobulin (IgG) and protein A from Staphylococcus aureus based on the immobilization of human IgG on the surface of modified gold-coated magnetic nanoparticles. The nanoparticles with an Au shell and Fe oxide cores were functionalized by a self-assembled monolayer of 11-mercaptoundecanoic acid. The electrochemical analysis was conducted on the modified magnetic carbon paste electrodes with the nanoparticles. The magnetic nanoparticles were attached to the surface of the magnetic carbon paste electrodes via magnetic force. The cyclic voltammetry technique and electrochemical impedance spectroscopy measurements of the magnetic carbon paste electrodes coated with magnetic nanoparticles-human IgG complex showed changes in its alternating current (AC) response both after the modification of the surface of the electrode and the addition of protein A. The immunological interaction between human IgG on the surface of the modified magnetic carbon paste electrodes and protein A in the solution could be successfully monitored.

Novel redox species polyaniline derivative-Au/Pt as sensing platform for label-free electrochemical immunoassay of carbohydrate antigen 199

Energy Technology Data Exchange (ETDEWEB)

Wang, Liyuan; Shan, Jiao; Feng, Feng; Ma, Zhanfang, E-mail: mazhanfang@cnu.edu.cn

2016-03-10

A novel electrochemical redox-active nanocomposite was synthesized by a one-pot method using N,N′-diphenyl-p-phenylediamine as monomer, and HAuCl{sub 4} and K{sub 2}PtCl{sub 4} as co-oxidizing agents. The as-prepared poly(N,N′-diphenyl-p-phenylediamine)-Au/Pt exhibited admirable electrochemical redox activity at 0.15 V, excellent H{sub 2}O{sub 2} electrocatalytic ability and favorable electron transfer ability. Based on these, the evaluation of the composite as sensing substrate for label-free electrochemical immunosensing to the sensitive detection of carbohydrate antigen 199 was described. This technique proved to be a prospective detection tool with a wide liner range from 0.001 U mL{sup −1} to 40 U mL{sup −1}, and a low detection limit of 2.3 × 10{sup −4} U mL{sup −1} (S/N = 3). In addition, this method was used for the analysis of human serum sample, and good agreement was obtained between the values and those of enzyme-linked immunosorbent assay, implying the potential application in clinical research. Importantly, the strategy of the present substrate could be extended to other polymer-based nanocomposites such as polypyrrole derivatives or polythiophene derivatives, and this could be of great significance for the electrochemical immunoassay. - Highlights: • A novel electrochemical redox composite PPPD-Au/Pt was synthesized by one-pot method. • PPPD-Au/Pt was used as sensing substrate for label-free electrochemical immunosensor. • The immunosensor showed wide detection range and ultralow detection limit for the detection of CA199.

The fabrication of magnetic particle-based chemiluminescence immunoassay for human epididymis protein-4 detection in ovarian cancer.

Science.gov (United States)

Fu, Xiaoling; Liu, Yangyang; Qiu, Ruiyun; Foda, Mohamed F; Zhang, Yong; Wang, Tao; Li, Jinshan

2018-03-01

The magnetic particles have a significant influence on the immunoassay detection and cancer therapy. Herein, the chemiluminescence immunoassay combined with the magnetic particles (MPCLIA) was presented for the clinical determination and analysis of human epididymis protein 4 (HE4) in the human serum. Under the optimized experiment conditions, the secure MPCLIA method can detect HE4 in the broader range of 0-1000 pmol/L, with a lower detection limit of 1.35 pmol/L. The satisfactory recovery rate of the method in the serum ranged from 83.62% to 105.10%, which was well within the requirement of clinical analysis. Moreover, the results showed the good correlation with enzyme-linked immunosorbent assay (ELISA), with the correlation coefficient of 0.9589. This proposed method has been successfully applied to the clinical determination of HE4 in the human serum.

Detection of alpha-fetoprotein in magnetic immunoassay of thin channels using biofunctional nanoparticles

Science.gov (United States)

Tsai, H. Y.; Gao, B. Z.; Yang, S. F.; Li, C. S.; Fuh, C. Bor

2014-01-01

This paper presents the use of fluorescent biofunctional nanoparticles (10-30 nm) to detect alpha-fetoprotein (AFP) in a thin-channel magnetic immunoassay. We used an AFP model biomarker and s-shaped deposition zones to test the proposed detection method. The results show that the detection using fluorescent biofunctional nanoparticle has a higher throughput than that of functional microparticle used in previous experiments on affinity reactions. The proposed method takes about 3 min (versus 150 min of previous method) to detect 100 samples. The proposed method is useful for screening biomarkers in clinical applications, and can reduce the run time for sandwich immunoassays to less than 20 min. The detection limits (0.06 pg/ml) and linear ranges (0.068 pg/ml-0.68 ng/ml) of AFP using fluorescent biofunctional nanoparticles are the same as those of using functional microparticles within experimental errors. This detection limit is substantially lower and the linear range is considerably wider than those of enzyme-linked immunosorbent assay (ELISA) and other methods in sandwich immunoassay methods. The differences between this method and an ELISA in AFP measurements of serum samples were less than 12 %. The proposed method provides simple, fast, and sensitive detection with a high throughput for biomarkers.

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.

Electrochemical synthesis of CORE-shell magnetic nanowires

KAUST Repository

Ovejero, Jesú s G.; Bran, Cristina; Vidal, Enrique Vilanova; Kosel, Jü rgen; Morales, Marí a P.; Vazquez, Manuel

2015-01-01

(Fe, Ni, CoFe) @ Au core-shell magnetic nanowires have been synthesized by optimized two-step potentiostatic electrodeposition inside self-assembled nanopores of anodic aluminium templates. The optimal electrochemical parameters (e.g., potential

Electrochemical sensor based on magnetic molecularly imprinted nanoparticles modified magnetic electrode for determination of Hb.

Science.gov (United States)

Sun, Binghua; Ni, Xinjiong; Cao, Yuhua; Cao, Guangqun

2017-05-15

A fast and selective electrochemical sensor for determination of hemoglobin (Hb) was developed based on magnetic molecularly imprinted nanoparticles modified on the magnetic glassy carbon electrode. The nanoparticles Fe 3 O 4 @SiO 2 with a magnetic core and a molecularly imprinted shell had regular structures and good monodispersity. Hb could be determined directly by electrochemical oxidization with the modified electrode. A magnetic field increased electrochemical response to Hb by two times. Imprinting Hb on the surface of Fe 3 O 4 @SiO 2 shortened the response time within 7min. Under optimum conditions, the imprinting factor toward the non-imprinted sensor was 2.8, and the separation factor of Hb to horseradish peroxidase was 2.6. The oxidation peak current had a linear relationship with Hb concentration ranged from 0.005mg/ml to 0.1mg/ml with a detection limit (S/N =3) of 0.0010mg/ml. The sensors were successfully applied to analysis of Hb in whole blood samples with recoveries between 95.7% and 105%. Copyright © 2016 Elsevier B.V. All rights reserved.

Label-free electrochemical immunoassay for neuron specific enolase based on 3D macroporous reduced graphene oxide/polyaniline film.

Science.gov (United States)

Zhang, Qi; Li, Xiaoyan; Qian, Chunhua; Dou, Li; Cui, Feng; Chen, Xiaojun

2018-01-01

The content of neuron specific enolase (NSE) in serum is considered to be an essential indicator of small cell lung cancer (SCLC). Here, a novel label-free electrochemical immunoassay for the detection of NSE based on the three dimensionally macroporous reduced graphene oxide/polyaniline (3DM rGO/PANI) film has been proposed. The 3DM rGO/PANI film was constructed by electrochemical co-deposition of GO and aniline into the interspaces of a sacrificial silica opal template modified Au slice. During the co-deposition, GO was successfully reduced by aniline and PANI could be deposited on the surfaces of rGO sheets. The ratio of rGO and PANI in the composite was also optimized to achieve the maximum electrochemical performance. The 3DM rGO/PANI composite provided larger specific surface area for the antibody immobilization, exhibited enhanced conductivity for electron transfer, and more important was that PANI acted as the electroactive probe for indicating the NSE concentration. Under the optimal conditions, a linear current response of PANI to NSE concentration was obtained over 0.5 pg mL -1 -10.0 ng mL -1 with a detection limit of 0.1 pg mL -1 . Moreover, the immunosensor showed excellent selectivity, good stability, satisfactory reproducibility and regeneration, and was employed to detect NSE in clinical serum specimens. Copyright © 2017 Elsevier Inc. All rights reserved.

Magnetic immunoassay coupled with inductively coupled plasma mass spectrometry for simultaneous quantification of alpha-fetoprotein and carcinoembryonic antigen in human serum

Energy Technology Data Exchange (ETDEWEB)

Zhang, Xing; Chen, Beibei; He, Man; Zhang, Yiwen; Xiao, Guangyang; Hu, Bin, E-mail: binhu@whu.edu.cn

2015-04-01

The absolute quantification of glycoproteins in complex biological samples is a challenge and of great significance. Herein, 4-mercaptophenylboronic acid functionalized magnetic beads were prepared to selectively capture glycoproteins, while antibody conjugated gold and silver nanoparticles were synthesized as element tags to label two different glycoproteins. Based on that, a new approach of magnetic immunoassay-inductively coupled plasma mass spectrometry (ICP-MS) was established for simultaneous quantitative analysis of glycoproteins. Taking biomarkers of alpha-fetoprotein (AFP) and carcinoembryonic antigen (CEA) as two model glycoproteins, experimental parameters involved in the immunoassay procedure were carefully optimized and analytical performance of the proposed method was evaluated. The limits of detection (LODs) for AFP and CEA were 0.086 μg L{sup −1} and 0.054 μg L{sup −1} with the relative standard deviations (RSDs, n = 7, c = 5 μg L{sup −1}) of 6.5% and 6.2% for AFP and CEA, respectively. Linear range for both AFP and CEA was 0.2–50 μg L{sup −1}. To validate the applicability of the proposed method, human serum samples were analyzed, and the obtained results were in good agreement with that obtained by the clinical chemiluminescence immunoassay. The developed method exhibited good selectivity and sensitivity for the simultaneous determination of AFP and CEA, and extended the applicability of metal nanoparticle tags based on ICP-MS methodology in multiple glycoprotein quantifications. - Highlights: • 4-Mercaptophenylboronic acid functionalized magnetic beads were prepared and characterized. • ICP-MS based magnetic immunoassay approach was developed for quantification of glycoproteins. • AFP and CEA were quantified simultaneously with Au and Ag NPs as element tags. • The developed method exhibited good selectivity and sensitivity for target glycoproteins.

Sensitive and rapid immunoassay for parathyroid hormone using magnetic particle labels and magnetic actuation.

Science.gov (United States)

Dittmer, W U; de Kievit, P; Prins, M W J; Vissers, J L M; Mersch, M E C; Martens, M F W C

2008-09-30

A rapid method for the sensitive detection of proteins using actuated magnetic particle labels, which are measured with a giant magneto-resistive (GMR) biosensor, is described. The technique involves a 1-step sandwich immunoassay with no fluid replacement steps. The various assay binding reactions as well as the bound/free separation are entirely controlled by magnetic forces induced by electromagnets above and below the sensor chip. During the assay, particles conjugated with tracer antibodies are actuated through the sample for target capture, and rapidly brought to the sensor surface where they bind to immobilized capture antibodies. Weakly or unbound labels are removed with a magnetic force oriented away from the GMR sensor surface. For the measurement of parathyroid hormone (PTH), a detection limit in the 10 pM range is obtained with a total assay time of 15 min when 300 nm particles are used. The same sensitivity can be achieved in 5 min when 500 nm particles are used. If 500 nm particles are employed in a 15-minute assay, then 0.8 pM of PTH is detectable. The low sample volume, high analytical performance and high speed of the test coupled with the compact GMR biosensor make the system especially suitable for sensitive testing outside of laboratory environments.

Catechol-chitosan redox capacitor for added amplification in electrochemical immunoanalysis.

Science.gov (United States)

Yan, Kun; Liu, Yi; Guan, Yongguang; Bhokisham, Narendranath; Tsao, Chen-Yu; Kim, Eunkyoung; Shi, Xiao-Wen; Wang, Qin; Bentley, William E; Payne, Gregory F

2018-05-22

Antibodies are common recognition elements for molecular detection but often the signals generated by their stoichiometric binding must be amplified to enhance sensitivity. Here, we report that an electrode coated with a catechol-chitosan redox capacitor can amplify the electrochemical signal generated from an alkaline phosphatase (AP) linked immunoassay. Specifically, the AP product p-aminophenol (PAP) undergoes redox-cycling in the redox capacitor to generate amplified oxidation currents. We estimate an 8-fold amplification associated with this redox-cycling in the capacitor (compared to detection by a bare electrode). Importantly, this capacitor-based amplification is generic and can be coupled to existing amplification approaches based on enzyme-linked catalysis or magnetic nanoparticle-based collection/concentration. Thus, the capacitor should enhance sensitivities in conventional immunoassays and also provide chemical to electrical signal transduction for emerging applications in molecular communication. Copyright © 2018 Elsevier B.V. All rights reserved.

Magnetic, catalytic, EPR and electrochemical studies on binuclear ...

Indian Academy of Sciences (India)

Magnetic, catalytic, EPR and electrochemical studies on binuclear copper(II) complexes ... to the oxidation of 3,5-di--butylcatechol to the corresponding quinone. ... EPR spectral studies in methanol solvent show welldefined four hyperfine ...

Development of a PMMA Electrochemical Microfluidic Device for Carcinoembryonic Antigen Detection

Science.gov (United States)

Van Anh, Nguyen; Van Trung, Hoang; Tien, Bui Quang; Binh, Nguyen Hai; Ha, Cao Hong; Le Huy, Nguyen; Loc, Nguyen Thai; Thu, Vu Thi; Lam, Tran Dai

2016-05-01

In this study, a poly(methyl methacrylate) (PMMA) microfluidic device fabricated by an inexpensive CO2 laser etching system was developed for detection of carcino-embryonic antigens (CEA). The device was capable of working in continuous mode and was designed with the aid of numerical simulation. The detection of target CEA was based on immuno-assay via magnetic particles and electrochemical sensing. The as-prepared microfluidic can be used to detect CEA at the relatively low concentration of 150 pg mL-1. The device could be reused many times, since the capture and removal of magnetic particles in the assay could be manipulated by an external magnetic field. The proposed approach appears to be suitable for high-throughput and automated analysis of large biomolecules such as tumor markers and pathogens.

Progress in electrochemical synthesis of magnetic iron oxide nanoparticles

International Nuclear Information System (INIS)

Ramimoghadam, Donya; Bagheri, Samira; Hamid, Sharifah Bee Abd

2014-01-01

Recently, magnetic iron oxide particles have been emerged as significant nanomaterials due to its extensive range of application in various fields. In this regard, synthesis of iron oxide nanoparticles with desirable properties and high potential applications are greatly demanded. Therefore, investigation on different iron oxide phases and their magnetic properties along with various commonly used synthetic techniques are remarked and thoroughly described in this review. Electrochemical synthesis as a newfound method with unique advantages is elaborated, followed by design approaches and key parameters to control the properties of the iron oxide nanoparticles. Additionally, since the dispersion of iron oxide nanoparticles is as important as its preparation, surface modification issue has been a serious challenge which is comprehensively discussed using different surfactants. Despite the advantages of the electrochemical synthesis method, this technique has been poorly studied and requires deep investigations on effectual parameters such as current density, pH, electrolyte concentration etc. - Highlights: • IONPs are applied in chemical industries, medicine, magnetic storage etc. • Electrochemical synthesis (EC) is convenient, eco-friendly, selective and low-cost. • EC key factors are current density, pH, electrolyte concentration, electrode type. • Organic, inorganic and biological materials can be used to modify IONPs’ surface. • The physicochemical properties of IONPs can be controlled by adding surfactants

Porous silicon platform for optical detection of functionalized magnetic particles biosensing.

Science.gov (United States)

Ko, Pil Ju; Ishikawa, Ryousuke; Sohn, Honglae; Sandhu, Adarsh

2013-04-01

The physical properties of porous materials are being exploited for a wide range of applications including optical biosensors, waveguides, gas sensors, micro capacitors, and solar cells. Here, we review the fast, easy and inexpensive electrochemical anodization based fabrication porous silicon (PSi) for optical biosensing using functionalized magnetic particles. Combining magnetically labeled biomolecules with PSi offers a rapid and one-step immunoassay and real-time detection by magnetic manipulation of superparamagnetic beads (SPBs) functionalized with target molecules onto corresponding probe molecules immobilized inside nano-pores of PSi. We first give an introduction to electrochemical and chemical etching procedures used to fabricate a wide range of PSi structures. Next, we describe the basic properties of PSi and underlying optical scattering mechanisms that govern their unique optical properties. Finally, we give examples of our experiments that demonstrate the potential of combining PSi and magnetic beads for real-time point of care diagnostics.

Magnetic bead and gold nanoparticle probes based immunoassay for β-casein detection in bovine milk samples.

Science.gov (United States)

Li, Y S; Meng, X Y; Zhou, Y; Zhang, Y Y; Meng, X M; Yang, L; Hu, P; Lu, S Y; Ren, H L; Liu, Z S; Wang, X R

2015-04-15

In this work, a double-probe based immunoassay was developed for rapid and sensitive determination of β-casein in bovine milk samples. In the method, magnetic beads (MBs), employed as supports for the immobilization of anti-β-casein polyclonal antibody (PAb), were used as the capture probe. Colloidal gold nanoparticles (AuNPs), employed as a bridge for loading anti-β-casein monoclonal antibody (McAb) and horseradish peroxidase (HRP), were used as the amplification probe. The presence of β-casein causes the sandwich structures of MBs-PAb-β-casein-McAb-AuNPs through the interaction between β-casein and the anti-β-casein antibodies. The HRP, used as an enzymatic-amplified tracer, can catalytically oxidize the substrate 3,3',5,5'-tetramethylbenzidine (TMB), generating optical signals that are proportional to the quantity of β-casein. The linear range of the immunoassay was from 6.5 to 1520ngmL(-1). The limit of detection (LOD) was 4.8ngmL(-1) which was 700 times lower than that of MBs-antibody-HRP based immunoassay and 6-7 times lower than that from the microplate-antibody-HRP based assay. The recoveries of β-casein from bovine milk samples were from 95.0% to 104.3% that had a good correlation coefficient (R(2)=0.9956) with those obtained by an official standard Kjeldahl method. For higher sensitivity, simple sample pretreatment and shorter time requirement of the antigen-antibody reaction, the developed immunoassay demonstrated the viability for detection of β-casein in bovine milk samples. Copyright © 2014. Published by Elsevier B.V.

A sandwich-type electrochemical immunoassay for ultrasensitive detection of non-small cell lung cancer biomarker CYFRA21-1.

Science.gov (United States)

Zeng, Yan; Bao, Jing; Zhao, Yanan; Huo, Danqun; Chen, Mei; Qi, Yanli; Yang, Mei; Fa, Huanbao; Hou, Changjun

2018-04-01

Many studies confirm that the aberrant expression of Cytokeratin 19 fragment 21-1 (CYFRA21-1) is highly correlated with non-small cell lung cancer (NSCLC), especially for squamous cell carcinoma. Herein, we report a sandwich-type electrochemical immunosensor based on signal amplification strategy of multiple nanocomposites to test CYFRA21-1 selectively and sensitively. The proposed immunosensor fabricated by three-dimensional graphene (3D-G), chitosan (CS) and glutaraldehyde (GA) composite on the glass carbon electrode (GCE) with a large surface area is prepared to immobilize primary antibodies (Ab 1 ) and provide excellent conductivity. To further amplify the electrochemical signal, the trace tag on the foundation of gold nanoparticles (AuNPs) is coated with amino-functionalized carbon nanotube (MWCNT-NH 2 ) nanocomposite through thionine linking, which provides more amino groups to capture more horseradish peroxidase-labeled antibodies (HPR-Ab 2 ) and enhances the conductivity. Under optimal conditions, the developed immunosensor exhibits excellent analytical performance for the determination of CYFRA21-1 with a wide linear range from 0.1 to 150ng·mL -1 and a low detection limit (LOD) of 43pg·mL -1 . Furthermore, satisfactory results are obtained for the determination of CYFRA21-1 in real clinical serum samples, indicating the potential of the immunoassay to be applied in clinical analysis. Copyright © 2017 Elsevier B.V. All rights reserved.

Functionalized gold nanorod-based labels for amplified electrochemical immunoassay of E. coli as indicator bacteria relevant to the quality of dairy product.

Science.gov (United States)

Zhang, Xinai; Zhang, Fan; Zhang, Hongyin; Shen, Jianzhong; Han, En; Dong, Xiaoya

2015-01-01

In this paper, we report an amplified electrochemical immunoassay for Escherichia coli as indicator bacteria relevant to the quality of dairy product using the functionalized gold nanorod-based labels ({dAb-AuNR-FCA}). The {dAb-AuNR-FCA} labels were designed by exploiting silica-functionalized gold nanorods (AuNR@SiO2) as the carriers for immobilization of detection antibody (dAb) and ferrocenecarboxylic acid (FCA), in which dAb was used for recognition of E. coli and FCA tags served as signal-generating molecule. Greatly amplified signal was achieved in the sandwich-type immunoassay when enormous FCA linked to AuNR@SiO2. Compared with the commercially available {dAb-FCA}, the {dAb-AuNR-FCA} labels exhibited a better performance for E. coli assay due to the advantages of AuNR@SiO2 as carriers. Under optimal experimental conditions, it showed a linear relationship between the peak current of FCA and the logarithmic value of E. coli concentration ranging from 1.0×10(2) to 5.0×10(4) cfu mL(-1) with a detection limit of 60 cfu mL(-1) (S/N=3), and the electrochemical detection of E. coli could be achieved in 3h. Moreover, the proposed strategy was used to determine E. coli in dairy product (pure fresh milk, yogurt in shelf-life, and expired yogurt), and the recoveries of standard additions were in the range of 95.1-106%. This proposed strategy exhibited rapid response, high sensitivity and specificity for E. coli assay in dairy product, and could become a promising technique to estimate the quality of dairy product. Copyright © 2014 Elsevier B.V. All rights reserved.

Alzheimer's disease biomarkers detection in human samples by efficient capturing through porous magnetic microspheres and labelling with electrocatalytic gold nanoparticles

Czech Academy of Sciences Publication Activity Database

de la Escosura-Muniz, A.; Plichta, Zdeněk; Horák, Daniel; Merkoci, A.

2015-01-01

Roč. 67, 15 May (2015), s. 162-169 ISSN 0956-5663 R&D Projects: GA MŠk 7E12053 EU Projects: European Commission(XE) 246513 - NADINE Institutional support: RVO:61389013 Keywords : porous magnetic microspheres * gold nanoparticles * electrochemical immunoassay Subject RIV: CD - Macromolecular Chemistry Impact factor: 7.476, year: 2015

Molecularly Imprinted Polymer as an Antibody Substitution in Pseudo-immunoassays for Chemical Contaminants in Food and Environmental Samples.

Science.gov (United States)

Chen, Chaochao; Luo, Jiaxun; Li, Chenglong; Ma, Mingfang; Yu, Wenbo; Shen, Jianzhong; Wang, Zhanhui

2018-03-21

The chemical contaminants in food and the environment are quite harmful to food safety and human health. Rapid, accurate, and cheap detection can effectively control the potential risks derived from these chemical contaminants. Among all detection methods, the immunoassay based on the specific interaction of antibody-analyte is one of the most widely used techniques in the field. However, biological antibodies employed in the immunoassay usually cannot tolerate extreme conditions, resulting in an unstable state in both physical and chemical profiles. Molecularly imprinted polymers (MIPs) are a class of polymers with specific molecular recognition abilities, which are highly robust, showing excellent operational stability under a wide variety of conditions. Recently, MIPs have been used in biomimetic immunoassays for chemical contaminants as an antibody substitute in food and the environment. Here, we reviewed these applications of MIPs incorporated in different analytical platforms, such as enzyme-linked immunosorbent assay, fluorescent immunoassay, chemiluminescent immunoassay, electrochemical immunoassay, microfluidic paper-based immunoassay, and homogeneous immunoassay, and discussed current challenges and future trends in the use of MIPs in biomimetic immunoassays.

A new strategy for label-free electrochemical immunoassay based on “gate-effect” of β-cyclodextrin modified electrode

Energy Technology Data Exchange (ETDEWEB)

Deng, Huan [College of Chemistry and Bioengineering, Guilin University of Technology, Guilin 541004 (China); Li, Jianping, E-mail: likianping@263.net [College of Chemistry and Bioengineering, Guilin University of Technology, Guilin 541004 (China); Zhang, Yun; Pan, Hongcheng [College of Chemistry and Bioengineering, Guilin University of Technology, Guilin 541004 (China); Xu, Guobao, E-mail: guobaoxu@ciac.ac.cn [College of Chemistry and Bioengineering, Guilin University of Technology, Guilin 541004 (China); State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022 (China)

2016-07-05

A novel label-free electrochemical immunoassay was developed for prostate-specific antigen (PSA) detection via using β-cyclodextrin (β-CD) assembled layer created gates for the electron transfer of probe. To construct the sensor, a gold electrode was self-assembled with monoclonal anti-PSA antibody labeled 6-mercapto-β-cyclodextrin. Interspaces among β-CD molecules in the layer were automatically formed on gold electrode, which act as the channel of the electron transfer of [Fe(CN){sub 6}]{sup 3−/4−} probe. When PSA bind with anti-PSA, it can block these channels on the electrode surface due to their steric hindrance effect, resulting in the decrease in redox current of the probe. Through such a gate-controlled effect, ultra trace amount of PSA may make the currents change greatly after the immunoreaction, which enhanced the signal-to-noise ratio to achieve the amplification effect. By evaluating the logarithm of PSA concentrations, the immunosensor had a good linear response to the current changes with a detection limit of 0.3 pg/mL (S/N = 3) when PSA concentration ranged from 1.0 pg/mL to 1.0 ng/mL. The label-free immunosensor exhibited satisfactory performances in sensitivity, repeatability as well as specificity. - Highlights: • A label-free PSA immunoassay was developed based on “gate-effect” amplification. • Interspaces among β-CD assembled for [Fe(CN){sub 6}]{sup 3−/4−} electron transfer were controlled by the immunoreaction. • Higher sensitivity was achieved with time and cost saving principle.

Magnetic field-assisted electrochemical discharge machining

International Nuclear Information System (INIS)

Cheng, Chih-Ping; Mai, Chao-Chuang; Wu, Kun-Ling; Hsu, Yu-Shan; Yan, Biing-Hwa

2010-01-01

Electrochemical discharge machining (ECDM) is an effective unconventional method for micromachining in non-conducting materials, such as glass, quartz and some ceramics. However, since the spark discharge performance becomes unpredictable as the machining depth increases, it is hard to achieve precision geometry and efficient machining rate in ECDM drilling. One of the main factors for this is the lack of sufficient electrolyte flow in the narrow gap between the tool and the workpiece. In this study a magnetohydrodynamic (MHD) convection, which enhances electrolyte circulation has been applied to the ECDM process in order to upgrade the machining accuracy and efficiency. During electrolysis in the presence of a magnetic field, the Lorenz force induces the charged ions to form a MHD convection. The MHD convection then forces the electrolyte into movement, thus enhancing circulation of electrolyte. Experimental results show that the MHD convection induced by the magnetic field can effectively enhance electrolyte circulation in the micro-hole, which contributes to higher machining efficiency. Micro-holes in glass with a depth of 450 µm are drilled in less than 20 s. At the same time, better electrolyte circulation can prevent deterioration of gas film quality with increasing machining depth, while ensuring stable electrochemical discharge. The improvement in the entrance diameter thus achieved was 23.8% while that in machining time reached 57.4%. The magnetic field-assisted approach proposed in the research does not require changes in the machining setup or electrolyte but has proved to achieve significant enhancement in both accuracy and efficiency of ECDM.

New generation of electrochemical immunoassay based on polymeric nanoparticles for early detection of breast cancer

Directory of Open Access Journals (Sweden)

Mouffouk F

2017-04-01

Full Text Available Fouzi Mouffouk,1 Sihem Aouabdi,2 Entesar Al-Hetlani,1 Hacene Serrai,3 Tareq Alrefae,4 Liaohai Leo Chen5 1Department of Chemistry, Kuwait University, Safat, Kuwait; 2King Abdullah International Medical Research Center (KAIMRC, Jeddah, Kingdom of Saudi Arabia; 3Department of Radiology and Nuclear Medicine, University Hospital of Gent (UZG, Gent, Belgium; 4Department of Physics, Kuwait University, Safat, Kuwait; 5Surgical Precision Research Lab. Department of Surgery, University of Illinois at Chicago, IL, USA Abstract: Screening and early diagnosis are the key factors for the reduction of mortality rate and treatment cost of cancer. Therefore, sensitive and selective methods that can reveal the low abundance of cancer biomarkers in a biological sample are always desired. Here, we report the development of a novel electrochemical biosensor for early detection of breast cancer by using bioconjugated self-assembled pH-responsive polymeric micelles. The micelles were loaded with ferrocene molecules as “tracers” to specifically target cell surface-associated epithelial mucin (MUC1, a biomarker for breast and other solid carcinoma. The synthesis of target-specific, ferrocene-loaded polymeric micelles was confirmed, and the resulting sensor was capable of detecting the presence of MUC1 in a sample containing about 10 cells/mL. Such a high sensitivity was achieved by maximizing the loading capacity of ferrocene inside the polymeric micelles. Every single event of binding between the antibody and antigen was represented by the signal of hundreds of thousands of ferrocene molecules that were released from the polymeric micelles. This resulted in a significant increase in the intensity of the ferrocene signal detected by cyclic voltammetry. Keywords: electrochemical immunoassay, polymeric nanoparticles, breast cancer biomarkers, biosensors 

Magnetic immunoassay coupled with inductively coupled plasma mass spectrometry for simultaneous quantification of alpha-fetoprotein and carcinoembryonic antigen in human serum

Science.gov (United States)

Zhang, Xing; Chen, Beibei; He, Man; Zhang, Yiwen; Xiao, Guangyang; Hu, Bin

2015-04-01

The absolute quantification of glycoproteins in complex biological samples is a challenge and of great significance. Herein, 4-mercaptophenylboronic acid functionalized magnetic beads were prepared to selectively capture glycoproteins, while antibody conjugated gold and silver nanoparticles were synthesized as element tags to label two different glycoproteins. Based on that, a new approach of magnetic immunoassay-inductively coupled plasma mass spectrometry (ICP-MS) was established for simultaneous quantitative analysis of glycoproteins. Taking biomarkers of alpha-fetoprotein (AFP) and carcinoembryonic antigen (CEA) as two model glycoproteins, experimental parameters involved in the immunoassay procedure were carefully optimized and analytical performance of the proposed method was evaluated. The limits of detection (LODs) for AFP and CEA were 0.086 μg L- 1 and 0.054 μg L- 1 with the relative standard deviations (RSDs, n = 7, c = 5 μg L- 1) of 6.5% and 6.2% for AFP and CEA, respectively. Linear range for both AFP and CEA was 0.2-50 μg L- 1. To validate the applicability of the proposed method, human serum samples were analyzed, and the obtained results were in good agreement with that obtained by the clinical chemiluminescence immunoassay. The developed method exhibited good selectivity and sensitivity for the simultaneous determination of AFP and CEA, and extended the applicability of metal nanoparticle tags based on ICP-MS methodology in multiple glycoprotein quantifications.

Ultrasensitive multi-analyte electrochemical immunoassay based on GNR-modified heated screen-printed carbon electrodes and PS@PDA-metal labels for rapid detection of MMP-9 and IL-6.

Science.gov (United States)

Shi, Jian-Jun; He, Ting-Ting; Jiang, Fang; Abdel-Halim, E S; Zhu, Jun-Jie

2014-05-15

An ultrasensitive electrochemical immunoassay was developed for rapid detection of interleukin-6 (IL-6) and matrix metallopeptidase-9 (MMP-9); the method utilized PS@PDA-metal nanocomposites based on graphene nanoribbon (GNR)-modified heated screen-printed carbon electrode (HSPCE). Because of the good hydrophilicity and low toxicity, GNRs were used to immobilize antibodies (Ab) and amplify the electrochemical signal. PS@PDA-metal was used to label antibodies and generate a strong electrochemical signal in acetic buffer. A sandwich strategy was adopted to achieve simultaneous detection of MMP-9 and IL-6 based on HSPCE without cross-talk between adjacent electrodes in the range of 10(-5) to 10(3) ng mL(-1) with detection limits of 5 fg mL(-1) and 0.1 pg mL(-1) (S/N=3), respectively. The proposed method showed wide detection range, low detection limit, acceptable stability and good reproducibility. Satisfactory results were also obtained in the practical samples, thus showing this is a promising technique for simultaneous clinical detection of biocomponent proteins. © 2013 Elsevier B.V. All rights reserved.

Magnetophoretic separation ICP-MS immunoassay using Cs-doped multicore magnetic nanoparticles for the determination of salmonella typhimurium.

Science.gov (United States)

Jeong, Arong; Lim, H B

2018-02-01

In this work, a magnetophoretic separation ICP-MS immunoassay using newly synthesized multicore magnetic nanoparticles (MMNPs) was developed for the determination of salmonella typhimurium (typhi). The uniqueness of this method was the use of MMNPs doped with Cs for both separation and detection, which enable us to achieve fast analysis, high sensitivity, and good reliability. For demonstration, heat-killed typhi in a phosphate buffer solution was determined by ICP-MS after the MMNP-typhi reaction product was separated from unreacted MMNPs in a micropipette tip filled with 25% polyethylene glycol through magnetophoretic separation. The calibration curve obtained by plotting 133 Cs intensity vs. the number of synthetic standard, showed a coefficient of determination (R 2 ) of 0.94 with a limit of detection (LOD) of 102 cells/mL without cell culturing. Excellent recoveries, between 98-100%, were obtained from four replicates and compared with a sandwich-type ICP-MS immunoassay for further confirmation. Copyright © 2017 Elsevier B.V. All rights reserved.

Magnetic beads-based electrochemical immunosensor for monitoring allergenic food proteins

Czech Academy of Sciences Publication Activity Database

Čadková, M.; Metelka, R.; Holubová, L.; Horák, Daniel; Dvořáková, V.; Bílková, Z.; Korecká, L.

2015-01-01

Roč. 484, 1 September (2015), s. 4-8 ISSN 0003-2697 R&D Projects: GA ČR GAP206/12/0381 Institutional support: RVO:61389013 Keywords : electrochemical immunosensor * magnetic particles * ovalbumin Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 2.243, year: 2015

In situ monitoring magnetism and resistance of nanophase platinum upon electrochemical oxidation

Directory of Open Access Journals (Sweden)

Eva-Maria Steyskal

2013-06-01

Full Text Available Controlled tuning of material properties by external stimuli represents one of the major topics of current research in the field of functional materials. Electrochemically induced property tuning has recently emerged as a promising pathway in this direction making use of nanophase materials with a high fraction of electrode-electrolyte interfaces. The present letter reports on electrochemical property tuning of porous nanocrystalline Pt. Deeper insight into the underlying processes could be gained by means of a direct comparison of the charge-induced response of two different properties, namely electrical resistance and magnetic moment. For this purpose, four-point resistance measurements and SQUID magnetometry were performed under identical in situ electrochemical control focussing on the regime of electrooxidation. Fully reversible variations of the electrical resistance and the magnetic moment of 6% and 1% were observed upon the formation or dissolution of a subatomic chemisorbed oxygen surface layer, respectively. The increase of the resistance, which is directly correlated to the amount of deposited oxygen, is considered to be primarily caused by charge-carrier scattering processes at the metal–electrolyte interfaces. In comparison, the decrease of the magnetic moment upon positive charging appears to be governed by the electric field at the nanocrystallite–electrolyte interfaces due to spin–orbit coupling.

Selective electrochemical extraction of REEs from NdFeB magnet waste at room temperature

NARCIS (Netherlands)

Venkatesan, P.; Vander Hoogerstraete, Tom; Hennebel, Tom; Binnemans, Koen; Sietsma, J.; Yang, Y.

2018-01-01

NdFeB magnet waste is one of the important secondary resources from which rare-earth elements (REEs) can be recovered. Herein we present an electrochemical route to selectively extract REEs from the magnet waste at room temperature. First, the magnet waste was partially leached with HCl. The

Preparation, characterization of Fe3O4 at TiO2 magnetic nanoparticles and their application for immunoassay of biomarker of exposure to organophosphorus pesticides

Energy Technology Data Exchange (ETDEWEB)

Zhang, Xiao; Wang, Hongbo; Yang, Chunming; Du, Dan; Lin, Yuehe

2013-03-15

Novel Fe3O4 at TiO2 magnetic nanoparticles were prepared and developed for a new nanoparticle-based immunosensor for electrochemical quantification of organophosphorylated butyrylcholinesterase (BChE) in plasma, a specific biomarker of exposure to organophosphorus (OP) agents. The Fe3O4 at TiO2 nanoparticles were synthesized by hydrolysis of tetrabutyltitanate on the surface of Fe3O4 magnetic nanospheres, and characterized by attenuated total reflection Fourier-transform infrared spectra, transmission electron microscope and X-ray diffraction. The functional Fe3O4 at TiO2 nanoparticles were performed as capture antibody to selectively enrich phosphorylated moiety instead of phosphoserine antibody in the traditional sandwich immunoassays. The secondary recognition was served by quantum dots (QDs)-tagged anti-BChE antibody (QDs-anti-BChE). With the help of a magnet, the resulting sandwich-like complex, Fe3O4 at TiO2/OP-BChE/QDs-anti-BChE, was easily isolated from sample solutions and the released cadmium ions were detected on a disposable screen-printed electrode (SPE). The binding affinities were investigated by both surface plasmon resonance (SPR) and square wave voltammetry (SWV). This method not only avoids the drawback of unavailability of commercial OP-specific antibody but also amplifies detection signal by QDs-tags together with easy separation of samples by magnetic forces. The proposed immunosensor yields a linear response over a broad OP-BChE concentrations range from 0.02 to 10 nM, with detection limit of 0.01 nM. Moreover, the disposable nanoparticle-based immunosensor has been validated with human plasma samples. It offers a new method for rapid, sensitive, selective and inexpensive screening/evaluating exposure to OP pesticides.

Electrochemical synthesis of magnetic nanostructures using anodic aluminum oxide templates

Science.gov (United States)

Gong, Jie

In this dissertation, template electrodeposition was employed to fabricate high quality magnetic nanostructures suited for the reliable investigation of novel spintronics phenomena such as CIMS, BMR, and CPP-GMR. Several critical aspects/steps relating to the synthesis process were investigated in this work. In order to obtain high quality magnetic nanostructures, free-standing and Si-supported anodic aluminum oxide templates with closely controlled pore diameters, lengths, as well as constriction sizes, were synthesized by anodization, followed by appropriate post-processing. The pore opening size on the barrier layer can be controlled down to 5 nm by ion beam etching. After optimization of the compositional, structural, and magnetic properties of homogeneous FeCoNiCu layers electrodeposited under different conditions, the pulsed deposition process of FeCoNI/Cu multilayers on n-Si was studied. The influence of Cu deposition potential and Fe2+ concentration on microstructure, chemical and electrochemical properties, magnetic properties, and hence magnetotransport properties were assessed. The dissolution of the FM layer during potential transition was minimized in order to control interface sharpness. Combined with the systematic sublayer thickness and FM layer composition optimization, unprecedented GMR sensitivity of 0.11%/Oe at 5-15 Oe was obtained. Growth of multilayer nanowires was performed, and contact to a single wire was attempted using an electrochemical technique. We succeeded in addressing a small number of nanowires and measured a CPP-GMR of 17%. Template electrodeposition thus provides a promising way to repeatably fabricate prototypes for spin dependent transport studies.

Non-enzymatic electrochemical immunoassay using noble metal nanoparticles: a review

International Nuclear Information System (INIS)

Tang, Juan; Tang, Dianping

2015-01-01

Electrochemical immunodetection has attracted considerable attention due to its high sensitivity, low cost and simplicity. Large efforts have recently made in order to design ultrasensitive assays. Noble metal nanoparticles (NM-NPs) offer advantages such as high conductivity and large surface-to-volume ratio. NM-NPs therefore are excellent candidates for developing electrochemical platforms for immunodetection and as signal tags. The use of biofunctionalized NM-NPs often results in amplified recognition via stronger loading of signal tags, and also in enhanced signal. This review (with 87 references) gives an overview on the current state in the use of NM-NPs in Non-enzymatic electrochemical immunosensing. We discuss the application of NM-NPs as electrode matrices and as electroactive labels (either as a carrier or as electrocatalytic labels), and compare the materials (mainly nanoparticles of gold, platinum, or of bimetallic materials) in terms of performance (for example by increasing sensitivity via label amplification or via high densities of capture molecules). A conclusion covers current challenges and gives an outlook. Rather than being exhaustive, the review focuses on representative examples that illustrate novel concepts and promising applications. NM-NPs based immunosensing opens a series of concepts for basic research and offers new tools for determination of trace amounts of protein-related analytes in environment and clinical applications. (author)

Direct electrochemistry and electrocatalysis of lobetyolin via magnetic functionalized reduced graphene oxide film fabricated electrochemical sensor

International Nuclear Information System (INIS)

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

2017-01-01

A novel lobetyolin electrochemical sensor based on a magnetic functionalized reduced graphene oxide/Nafion nanohybrid film has been introduced in this work. The magnetic functionalized reduced graphene oxide was characterized by fourier transform infrared spectroscopy, atomic force microscope, X-ray diffraction, transmission electron microscopy and thermogravimetric analysis. The scanning electron microscopy characterized the morphology and microstructure of the prepared sensors, and the electrochemical effective surface areas of the prepared sensors were also calculated by chronocoulometry method. The electrochemical behavior of lobetyolin on the magnetic functionalized reduced graphene oxide/Nafion nanohybrid modified glassy carbon electrode was investigated by cyclic voltammetry and differential pulse voltammetry in a phosphate buffer solution of pH 6.0. The electron-transfer coefficient (α), electron transfer number (n), and electrode reaction rate constant (Κs) were calculated as 0.78, 0.73, and 4.63 s −1 , respectively. Under the optimized conditions, the sensor based on magnetic functionalized reduced graphene oxide/Nafion showed a linear voltammetric response to the lobetyolin concentration at 1.0 × 10 −7 to 1.0 × 10 −4 mol/L with detection limit (S/N = 3)of 4.3 × 10 −8 mol/L. The proposed sensor also displayed acceptable reproducibility, long-term stability, and high selectivity, and performs well for analysis of lobetyolin in real samples. The voltammetric sensor was successfully applied to detect lobetyolin in Codonopsis pilosula with recovery values in the range of 96.12% –102.66%. - Graphical abstract: Schematic diagram of the synthesis of MrGO hybrid and the fabrication process of the MrGO/Nafion/GCE for determination of lobetyolin. Display Omitted - Highlights: • The MrGO/Nafion@GCE electrochemical sensor was successfully fabricated. • The prepared MrGO was characterized by AFM, XRD, FTIR, VSM, TEM and SEM. • The proposed

Direct electrochemistry and electrocatalysis of lobetyolin via magnetic functionalized reduced graphene oxide film fabricated electrochemical sensor

Energy Technology Data Exchange (ETDEWEB)

Sun, Bolu [School of Pharmacy, Lanzhou University, Lanzhou 730000 (China); Gou, Xiaodan [School of Chemistry and Chemical Engineering, Nanjing University, 210046 (China); Bai, Ruibin; Abdelmoaty, Ahmed Attia Ahmed; Ma, Yuling; Zheng, Xiaoping [School of Pharmacy, Lanzhou University, Lanzhou 730000 (China); Hu, Fangdi, E-mail: hufd@lzu.edu.cn [School of Pharmacy, Lanzhou University, Lanzhou 730000 (China)

2017-05-01

A novel lobetyolin electrochemical sensor based on a magnetic functionalized reduced graphene oxide/Nafion nanohybrid film has been introduced in this work. The magnetic functionalized reduced graphene oxide was characterized by fourier transform infrared spectroscopy, atomic force microscope, X-ray diffraction, transmission electron microscopy and thermogravimetric analysis. The scanning electron microscopy characterized the morphology and microstructure of the prepared sensors, and the electrochemical effective surface areas of the prepared sensors were also calculated by chronocoulometry method. The electrochemical behavior of lobetyolin on the magnetic functionalized reduced graphene oxide/Nafion nanohybrid modified glassy carbon electrode was investigated by cyclic voltammetry and differential pulse voltammetry in a phosphate buffer solution of pH 6.0. The electron-transfer coefficient (α), electron transfer number (n), and electrode reaction rate constant (Κs) were calculated as 0.78, 0.73, and 4.63 s{sup −1}, respectively. Under the optimized conditions, the sensor based on magnetic functionalized reduced graphene oxide/Nafion showed a linear voltammetric response to the lobetyolin concentration at 1.0 × 10{sup −7} to 1.0 × 10{sup −4} mol/L with detection limit (S/N = 3)of 4.3 × 10{sup −8} mol/L. The proposed sensor also displayed acceptable reproducibility, long-term stability, and high selectivity, and performs well for analysis of lobetyolin in real samples. The voltammetric sensor was successfully applied to detect lobetyolin in Codonopsis pilosula with recovery values in the range of 96.12% –102.66%. - Graphical abstract: Schematic diagram of the synthesis of MrGO hybrid and the fabrication process of the MrGO/Nafion/GCE for determination of lobetyolin. Display Omitted - Highlights: • The MrGO/Nafion@GCE electrochemical sensor was successfully fabricated. • The prepared MrGO was characterized by AFM, XRD, FTIR, VSM, TEM and SEM. �

Electrochemical genosensing of Salmonella, Listeria and Escherichia coli on silica magnetic particles

Energy Technology Data Exchange (ETDEWEB)

Liébana, Susana; Brandão, Delfina [Grup de Sensors i Biosensors, Departament de Química, Universitat Autònoma de Barcelona, 08193, Cerdanyola del Vallès (Bellaterra) (Spain); Cortés, Pilar; Campoy, Susana [Departament de Genètica i de Microbiologia, Universitat Autònoma de Barcelona, 08193, Cerdanyola del Vallès (Bellaterra) (Spain); Alegret, Salvador [Grup de Sensors i Biosensors, Departament de Química, Universitat Autònoma de Barcelona, 08193, Cerdanyola del Vallès (Bellaterra) (Spain); Pividori, María Isabel, E-mail: Isabel.Pividori@uab.cat [Grup de Sensors i Biosensors, Departament de Química, Universitat Autònoma de Barcelona, 08193, Cerdanyola del Vallès (Bellaterra) (Spain)

2016-01-21

A magneto-genosensing approach for the detection of the three most common pathogenic bacteria in food safety, such as Salmonella, Listeria and Escherichia coli is presented. The methodology is based on the detection of the tagged amplified DNA obtained by single-tagging PCR with a set of specific primers for each pathogen, followed by electrochemical magneto-genosensing on silica magnetic particles. A set of primers were selected for the amplification of the invA (278 bp), prfA (217 bp) and eaeA (151 bp) being one of the primers for each set tagged with fluorescein, biotin and digoxigenin coding for Salmonella enterica, Listeria monocytogenes and E. coli, respectively. The single-tagged amplicons were then immobilized on silica MPs based on the nucleic acid-binding properties of silica particles in the presence of the chaotropic agent as guanidinium thiocyanate. The assessment of the silica MPs as a platform for electrochemical magneto-genosensing is described, including the main parameters to selectively attach longer dsDNA fragments instead of shorter ssDNA primers based on their negative charge density of the sugar-phosphate backbone. This approach resulted to be a promising detection tool with sensing features of rapidity and sensitivity very suitable to be implemented on DNA biosensors and microfluidic platforms. - Highlights: • Silica magnetic particles were used for the first time as carrier in electrochemical magneto-genosensing of single-tagged amplicons. • They demonstrated to be a robust platform for the electrochemical detection of PCR products. • Differential adsorption properties for longer dsDNA amplicon incorporating the tagging primers over shorter ssDNA tagged primers were observed due to the negative charge density. • Electrochemical magneto-genosensing of Salmonella enterica, Listeria monocytogenes and Escherichia coli was successfully performed.

Electrochemical genosensing of Salmonella, Listeria and Escherichia coli on silica magnetic particles

International Nuclear Information System (INIS)

Liébana, Susana; Brandão, Delfina; Cortés, Pilar; Campoy, Susana; Alegret, Salvador; Pividori, María Isabel

2016-01-01

A magneto-genosensing approach for the detection of the three most common pathogenic bacteria in food safety, such as Salmonella, Listeria and Escherichia coli is presented. The methodology is based on the detection of the tagged amplified DNA obtained by single-tagging PCR with a set of specific primers for each pathogen, followed by electrochemical magneto-genosensing on silica magnetic particles. A set of primers were selected for the amplification of the invA (278 bp), prfA (217 bp) and eaeA (151 bp) being one of the primers for each set tagged with fluorescein, biotin and digoxigenin coding for Salmonella enterica, Listeria monocytogenes and E. coli, respectively. The single-tagged amplicons were then immobilized on silica MPs based on the nucleic acid-binding properties of silica particles in the presence of the chaotropic agent as guanidinium thiocyanate. The assessment of the silica MPs as a platform for electrochemical magneto-genosensing is described, including the main parameters to selectively attach longer dsDNA fragments instead of shorter ssDNA primers based on their negative charge density of the sugar-phosphate backbone. This approach resulted to be a promising detection tool with sensing features of rapidity and sensitivity very suitable to be implemented on DNA biosensors and microfluidic platforms. - Highlights: • Silica magnetic particles were used for the first time as carrier in electrochemical magneto-genosensing of single-tagged amplicons. • They demonstrated to be a robust platform for the electrochemical detection of PCR products. • Differential adsorption properties for longer dsDNA amplicon incorporating the tagging primers over shorter ssDNA tagged primers were observed due to the negative charge density. • Electrochemical magneto-genosensing of Salmonella enterica, Listeria monocytogenes and Escherichia coli was successfully performed.

Electrochemical immunoassay for the carcinoembryonic antigen based on the use of a glassy carbon electrode modified with an octahedral Cu2O-gold nanocomposite and staphylococcal protein for signal amplification.

Science.gov (United States)

Qin, Zhen; Xu, Wei; Chen, Shuai; Chen, Jun; Qiu, Jing Fu; Li, Chao Rui

2018-04-24

The authors describe an electrochemical immunoassay for ultrasensitive direct determination of the carcinoembryonic antigen (CEA). A nanocomposite consisting of octahedral Cu2O nanocrystals covered with gold nanoparticles was utilized to modify a glassy carbon electrode which gives a strongly enhanced chronoamperometric signal for H 2 O 2 which is used as an electrochemical probe. The morphology and elemental composition of the the nanocomposite was studied by field emission scanning electron microscopy and energy dispersive X-ray spectroscopy. In addition, staphylococcal protein A was placed on the electrode for efficient capture of antibody to further enhance the sensitivity of the assay. Under optimal conditions and at a typical working voltage of -0.4 V (vs. Ag/AgCl), the response covers the 2 pg·mL -1 to 20 ng·mL -1 CEA concentration range with a 200 fg·mL -1 lower detection limit. The method was successfully applied to the determination of CEA in (spiked) human serum. Graphical abstract Schematic of the fabrication of an electrochemical immunosensor for ultrasensitive detection the carcinoembryonic antigen. The sensor is based on the use of a glassy carbon electrode modified with an octahedral Cu 2 O-gold nanocomposite and staphylococcal protein A for signal amplification.

Detection of Hepatitis C core antibody by dual-affinity yeast chimera and smartphone-based electrochemical sensing.

Science.gov (United States)

Aronoff-Spencer, Eliah; Venkatesh, A G; Sun, Alex; Brickner, Howard; Looney, David; Hall, Drew A

2016-12-15

Yeast cell lines were genetically engineered to display Hepatitis C virus (HCV) core antigen linked to gold binding peptide (GBP) as a dual-affinity biobrick chimera. These multifunctional yeast cells adhere to the gold sensor surface while simultaneously acting as a "renewable" capture reagent for anti-HCV core antibody. This streamlined functionalization and detection strategy removes the need for traditional purification and immobilization techniques. With this biobrick construct, both optical and electrochemical immunoassays were developed. The optical immunoassays demonstrated detection of anti-HCV core antibody down to 12.3pM concentrations while the electrochemical assay demonstrated higher binding constants and dynamic range. The electrochemical format and a custom, low-cost smartphone-based potentiostat ($20 USD) yielded comparable results to assays performed on a state-of-the-art electrochemical workstation. We propose this combination of synthetic biology and scalable, point-of-care sensing has potential to provide low-cost, cutting edge diagnostic capability for many pathogens in a variety of settings. Copyright © 2016 Elsevier B.V. All rights reserved.

Rapid extraction and quantitative detection of the herbicide diuron in surface water by a hapten-functionalized carbon nanotubes based electrochemical analyzer.

Science.gov (United States)

Sharma, Priyanka; Bhalla, Vijayender; Tuteja, Satish; Kukkar, Manil; Suri, C Raman

2012-05-21

A solid phase extraction micro-cartridge containing a non-polar polystyrene absorbent matrix was coupled with an electrochemical immunoassay analyzer (EIA) and used for the ultra-sensitive detection of the phenyl urea herbicide diuron in real samples. The EIA was fabricated by using carboxylated carbon nanotubes (CNTs) functionalized with a hapten molecule (an amine functionalized diuron derivative). Screen printed electrodes (SPE) were modified with these haptenized CNTs and specific in-house generated anti diuron antibodies were used for bio-interface development. The immunodetection was realized in a competitive electrochemical immunoassay format using alkaline phosphatase labeled secondary anti-IgG antibody. The addition of 1-naphthyl phosphate substrate resulted in the production of an electrochemically active product, 1-naphthol, which was monitored by using differential pulse voltammetry (DPV). The assay exhibited excellent sensitivity and specificity having a dynamic response range of 0.01 pg mL(-1) to 10 μg mL(-1) for diuron with a limit of detection of around 0.1 pg mL(-1) (n = 3) in standard water samples. The micro-cartridge coupled hapten-CNTs modified SPE provided an effective and efficient electrochemical immunoassay for the real-time monitoring of pesticides samples with a very high degree of sensitivity.

Electrochemical sensing platform based on tris(2,2'-bipyridyl)cobalt(III) and multiwall carbon nanotubes-Nafion composite for immunoassay of carcinoma antigen-125

International Nuclear Information System (INIS)

Chen Shihong; Yuan Ruo; Chai Yaqin; Min Ligen; Li Wenjuan; Xu Yang

2009-01-01

A new strategy for constructing a sensitive mediator-type electrochemical immunosensor for the detection of carcinoma antigen-125 (CA125) was developed. In this strategy, mediator tris(2,2'-bipyridyl)cobalt(III) (Co(bpy) 3 3+ ) was incoporated into the multiwall carbon nanotubes-Nafion (MWNTs-Nafion) composite film via a simple ion-exchange route. Then, gold colloidal nanoparticles (nano-Au) were attached onto Co(bpy) 3 3+ /MWNTs-Nafion film through electrostatic interaction between negatively charged nano-Au and positively charged Co(bpy) 3 3+ . Finally, CA125 monoclonal antibody (anti-CA125), used as a model antibody, was assembled onto the surface of nano-Au to achieve an immunosensor for the determination of CA125 antigen. X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) were used to characterize the assembly process of the modified electrode. The resulting immunosensor showed a high sensitivity, wide dynamic range consisting of two linear parts from 1.0 to 30 U mL -1 and 30 to 150 U mL -1 with a low detection limit of 0.36 U mL -1 at 3 times the background noise. Moreover, it displayed good reproducibility and stability, and would be potentially attractive for clinical immunoassay of CA125. The integration of mediator Co(bpy) 3 3+ and MWNTs-Nafion composite would offer potential promise for the fabrication of biosensors and biocatalysts.

Electrochemical magneto-immunosensing of Salmonella based on nano and micro-sized magnetic particles

International Nuclear Information System (INIS)

Brandão, D; Liébana, S; Alegret, S; Pividori, M I; Campoy, S; Cortés, P

2013-01-01

A very simple and rapid method for the detection of S. enterica is reported. In this approach, the bacteria were captured and preconcentrated with magnetic particles through an immunological reaction. A second polyclonal antibody labeled with peroxidase was used for the electrochemical immunosensing based on a magneto-electrode. Different nano and micro-sized magnetic particles were evaluated in this approach. The 'IMS/m-GEC electrochemical immunosensing' system shows a limit of detection of 5×10 4 and 1×10 4 CFU mL −1 in BHI culturing media when micro and nanoparticles are used respectively. These LOD were achieved in a total assay time of 1 h without any previous culturing preenrichment step. Moreover, this system was able to clearly distinguish between food pathogenic bacteria such as S. enterica and E. coli. The features of this approach were discussed and compared with conventional culture methods.

Electrochemical fabrication, microstructure and magnetic properties of Sm2Co17/Fe7Co3 dual phase nanocomposite

International Nuclear Information System (INIS)

Cui, Chunxiang; Chen, Fenghua; Yang, Wei; Li, Hongfang; Liu, Qiaozhi; Sun, Jibing

2015-01-01

By utilizing alternate electrochemical reaction, atomic migration and deposition of Fe, Co, Sm and other chemical substances in the electrochemical solution, a large number of Sm 2 Co 17 /Fe 7 Co 3 dual phase nanowire arrays were carried out in the anodic aluminum oxide (AAO) template with highly uniform and orderly. The Sm 2 Co 17 /Fe 7 Co 3 dual phase nanowire arrays with diameter of 50 nm and length of 12 μm have the smooth surface and uniform diameter. The morphology and microstructure of annealed Sm 2 Co 17 /Fe 7 Co 3 dual phase nanowires were observed and analyzed using SEM, TEM and HRTEM. Compared with single-phase nanowires, dual phase magnetic nanowires have higher coercivity and saturation magnetization. In this composite system, both the hard and the soft phases have a high Curie temperature, therefore, we believe that the Sm 2 Co 17 /Fe 7 Co 3 dual phase nanowire arrays is a new type of high-temperature magnetic composites. - Highlights: • Sm 2 Co 17 /Fe 7 Co 3 dual phase nanowires were prepared by electrochemical method. • The interface pinning is the main factor to improve anisotropy field of the nanowires. • The dual phase magnetic nanowires have higher coercivity and saturation magnetization

Immunoassays in Biotechnology

Science.gov (United States)

Immunoassays have broad applications for a wide variety of important biological compounds and environmental contaminants. Immunoassays can detect the presence of an antigen in the human body, a pollutant in the environment, or a critical antibody in a patient’s serum to develop a...

Disposable Electrochemical Immunosensor Diagnosis Device Based on Nanoparticle Probe and Immunochromatographic Strip

Energy Technology Data Exchange (ETDEWEB)

Liu, Guodong; Lin, Ying-Ying; Wang, Jun; Wu, Hong; Wai, Chien M.; Lin, Yuehe

2007-10-15

We describe a disposable electrochemical immunosensor diagnosis device that is based on the immunochromatographic strip technique and an electrochemical immunoassay based on quantum dot (QD, CdS@ZnS) labels. The device takes advantage of the speed and low-cost of the conventional immunochromatographic strip test and the high-sensitivity of the nanoparticle-based electrochemical immunoassay. A sandwich immunoreaction was performed on the immunochromatographic strip, and the captured QD labels in the test zone were determined by highly sensitive stripping voltammetric measurement of the dissolved metallic component (cadmium) with a disposable-screen-printed electrode, which is embedded underneath the membrane on the test zone. The new device coupled with a portable electrochemical analyzer shows great promise for in-field and point-of-care quantitative testing of disease-related protein biomarkers. The parameters (e.g., voltammetric measurement of QD labels, antibody immobilization, the loading amount of QD-antibody, and the immunoreaction time) that govern the sensitivity and reproducibility of the device were optimized with IgG model analyte. The voltammetric response of the optimized device is highly linear over the range of 0.1 to 10 ng mL-1 IgG, and the limit of detection is estimated to be 30 pg mL-1 in association with a 7-min immunoreaction time. The detection limit was improved to 10 pg mL-1 using a 20-min immunoreaction time. The new disposable electrochemical diagnosis device thus provides a more user-friendly, rapid, clinically accurate, less expensive, and quantitative tool for protein detection.

Specificity of immunoassays. Pt. 2

International Nuclear Information System (INIS)

Pratt, J.J.; Woldring, M.G.; Boonman, R.; Kittikool, J.

1979-01-01

Practical aspects of the measurement of the specificity of immunoassay are reviewed. Antibody heterogeneity in an antiserum makes a pragmatic rather than a theoretical approach necessary. A new method for the measurement of immunoassay specificity is described. This method is based on the errors caused by the cross-reacting antigens and is directly relevant to the validity of results obtained by immunoassay methods. The effect of selectively blocking the least specific antibodies in antisera raised against steroid haptens is tested. The practical consequences of these considerations are tested using steroid radioimmunoassay and enzyme-immunoassay. (orig.) [de

Flotation Immunoassay: Masking the Signal from Free Reporters in Sandwich Immunoassays.

Science.gov (United States)

Chen, Hui; Hagström, Anna E V; Kim, Jinsu; Garvey, Gavin; Paterson, Andrew; Ruiz-Ruiz, Federico; Raja, Balakrishnan; Strych, Ulrich; Rito-Palomares, Marco; Kourentzi, Katerina; Conrad, Jacinta C; Atmar, Robert L; Willson, Richard C

2016-04-14

In this work, we demonstrate that signal-masking reagents together with appropriate capture antibody carriers can eliminate the washing steps in sandwich immunoassays. A flotation immunoassay (FI) platform was developed with horseradish peroxidase chemiluminescence as the reporter system, the dye Brilliant Blue FCF as the signal-masking reagent, and buoyant silica micro-bubbles as the capture antibody carriers. Only reporters captured on micro-bubbles float above the dye and become visible in an analyte-dependent manner. These FIs are capable of detecting proteins down to attomole levels and as few as 10(6) virus particles. This signal-masking strategy represents a novel approach to simple, sensitive and quantitative immunoassays in both laboratory and point-of-care settings.

A biofilm microreactor system for simultaneous electrochemical and nuclear magnetic resonance techniques

International Nuclear Information System (INIS)

Renslow, Ryan S.; Babauta, Jerome T.; Majors, Paul D.; Mehta, Hardeep S.; Ewing, R. James; Ewing, Thomas; Mueller, Karl T.; Beyenal, Haluk

2014-01-01

In order to fully understand electrochemically active biofilms and the limitations to their scale-up in industrial biofilm reactors, a complete picture of the microenvironments inside the biofilm is needed. Nuclear magnetic resonance (NMR) techniques are ideally suited for the study of biofilms and for probing their microenvironments because these techniques allow for non-invasive interrogation and in situ monitoring with high resolution. By combining NMR with simultaneous electrochemical techniques, it is possible to sustain and study live electrochemically active biofilms. Here, we introduce a novel biofilm microreactor system that allows for simultaneous electrochemical and NMR techniques (EC-NMR) at the microscale. Microreactors were designed with custom radiofrequency resonator coils, which allowed for NMR measurements of biofilms growing on polarized gold electrodes. For an example application of this system, we grew Geobacter sulfurreducens biofilms. NMR was used to investigate growth media flow velocities, which were compared to simulated laminar flow, and electron donor concentrations inside the biofilms. We use Monte Carlo error analysis to estimate standard deviations of the electron donor concentration measurements within the biofilm. The EC-NMR biofilm microreactor system can ultimately be used to correlate extracellular electron transfer rates with metabolic reactions and explore extracellular electron transfer mechanisms

Magnetic-field-assisted synthesis of Co{sub 3}O{sub 4} nanoneedles with superior electrochemical capacitance

Energy Technology Data Exchange (ETDEWEB)

Zhu, Tao; Xie, Yan; Zhang, Guoxiong; He, Zhenni; Lu, Yisheng; Guo, Haibo [Shanghai University, Department of Electronic Information Materials, School of Materials Science and Engineering (China); Lin, Chuan [GE Global Research, China Technology Center (China); Chen, Yigang, E-mail: yigangchen@shu.edu.cn [Shanghai University, Department of Electronic Information Materials, School of Materials Science and Engineering (China)

2015-12-15

Nanostructured Co{sub 3}O{sub 4} films have been deposited on nickel foam in a magnetic-field-assisted hydrothermal process followed by annealing in air. The magnetic field strength is varied to study its relationship with nanostructures, morphology, and electrochemical properties of the Co{sub 3}O{sub 4} electrodes. The Co{sub 3}O{sub 4} films synthesized in the weak magnetic fields consist of dispersed nanoneedles, which are different from clustered nanoneedles when the magnetic field is absent. Moreover, the magnetic fields (of several millitesla) induced substantial changes in the nanostructures and electrochemical properties of the Co{sub 3}O{sub 4} films. A possible formation mechanism of Co{sub 3}O{sub 4} nanoneedles is proposed by comparing the morphologies and nanostructures of the films synthesized with and without the magnetic fields. Among these electrodes, the optimal one has a high specific capacitance (970.8 F g{sup −1} at 0.5 A g{sup −1}), good power capability (847.5 F g{sup −1} at 6.0 A g{sup −1}), and an excellent retention ratio (93.7 % over 1000 cycles). All these impressive results demonstrate that magnetic fields may be an economic and effective tool in hydrothermal synthesis of Co{sub 3}O{sub 4} electrodes for high-performance supercapacitors.

A magnetic particles-based chemiluminescence enzyme immunoassay for rapid detection of ovalbumin.

Science.gov (United States)

Feng, Xiao-Li; Ren, Hong-Lin; Li, Yan-Song; Hu, Pan; Zhou, Yu; Liu, Zeng-Shan; Yan, Dong-Ming; Hui, Qi; Liu, Dong; Lin, Chao; Liu, Nan-Nan; Liu, Yan-Yan; Lu, Shi-Ying

2014-08-15

Egg allergy is an important public health and safety concern, so quantification and administration of food or vaccines containing ovalbumin (OVA) are urgently needed. This study aimed to establish a rapid and sensitive magnetic particles-chemiluminescence enzyme immunoassay (MPs-CLEIA) for the determination of OVA. The proposed method was developed on the basis of a double antibodies sandwich immunoreaction and luminol-H2O2 chemiluminescence system. The MPs served as both the solid phase and separator, the anti-OVA MPs-coated polyclonal antibodies (pAbs) were used as capturing antibody, and the horseradish peroxidase (HRP)-labeled monoclonal antibody (mAb) was taken as detecting antibody. The parameters of the method were evaluated and optimized. The established MPs-CLEIA method had a linear range from 0.31 to 100ng/ml with a detection limit of 0.24ng/ml. The assays showed low reactivities and less than 5% of intraassay and interassay coefficients of variation (CVs), and the average recoveries were between 92 and 97%. Furthermore, the developed method was applied in real samples analysis successfully, and the correlation coefficient with the commercially available OVA kit was 0.9976. Moreover, it was more rapid and sensitive compared with the other methods for testing OVA. Copyright © 2014 Elsevier Inc. All rights reserved.

Immunoassay for serum amyloid A using a glassy carbon electrode modified with carboxy-polypyrrole, multiwalled carbon nanotubes, ionic liquid and chitosan

International Nuclear Information System (INIS)

Xia, Chunyong; Li, Yuan; Yuan, Guolin; Guo, Yanlei; Yu, Chao

2015-01-01

We report on a highly sensitive electrochemical immunoassay for the serum inflammation marker amyloid A (SAA). It is making use of a glassy carbon electrode that was modified with carboxy-endcapped polypyrrole (PPy-α-COOH), multiwalled carbon nanotubes (MWCNTs), ionic liquid and chitosan acting as the support platform. The nanocomposite increases the sensitivity and stability of the assay. Antibody against SAA was immobilized on a monolayer surface consisting of PPy-α-COOH. The electrode material was characterized by scanning electron microscopy, transmission electron microscopy, Fourier transform infrared spectra, cyclic voltammetry, electrochemical impedance spectroscopy and differential pulse voltammetry. The calibration plot for this assay, when operated at 0.16 V (vs. SCE) and applied to spiked serum samples, is linear in the 0.001 to 900 ng mL −1 SAA concentration range, and the detection limit is as low as 0.3 pg mL −1 (at an S/N ratio of 3). The electrode is stable and highly sensitive. The detection scheme is likely to be applicable to numerous other kinds of immunoassays. (author)

Immunoassay of paralytic shellfish toxins by moving magnetic particles in a stationary liquid-phase lab-on-a-chip.

Science.gov (United States)

Kim, Myoung-Ho; Choi, Suk-Jung

2015-04-15

In this study, we devised a stationary liquid-phase lab-on-a-chip (SLP LOC), which was operated by moving solid-phase magnetic particles in the stationary liquid phase. The SLP LOC consisted of a sample chamber to which a sample and reactants were added, a detection chamber containing enzyme substrate solution, and a narrow channel connecting the two chambers and filled with buffer. As a model system, competitive immunoassays of saxitoxin (STX), a paralytic shellfish toxin, were conducted in the SLP LOC using protein G-coupled magnetic particles (G-MPs) as the solid phase. Anti-STX antibodies, STX-horseradish peroxidase conjugate, G-MPs, and a STX sample were added to the sample chamber and reacted by shaking. While liquids were in the stationary state, G-MPs were transported from the sample chamber to the detection chamber by moving a magnet below the LOC. After incubation to allow the enzymatic reaction to occur, the absorbance of the detection chamber solution was found to be reciprocally related to the STX concentration of the sample. Thus, the SLP LOC may represent a novel, simple format for point-of-care testing applications of enzyme-linked immunosorbent assays by eliminating complicated liquid handling steps. Copyright © 2014 Elsevier B.V. All rights reserved.

Enzyme-linked electrochemical DNA ligation assay using magnetic beads.

Science.gov (United States)

Stejskalová, Eva; Horáková, Petra; Vacek, Jan; Bowater, Richard P; Fojta, Miroslav

2014-07-01

DNA ligases are essential enzymes in all cells and have been proposed as targets for novel antibiotics. Efficient DNA ligase activity assays are thus required for applications in biomedical research. Here we present an enzyme-linked electrochemical assay based on two terminally tagged probes forming a nicked junction upon hybridization with a template DNA. Nicked DNA bearing a 5' biotin tag is immobilized on the surface of streptavidin-coated magnetic beads, and ligated product is detected via a 3' digoxigenin tag recognized by monoclonal antibody-alkaline phosphatase conjugate. Enzymatic conversion of napht-1-yl phosphate to napht-1-ol enables sensitive detection of the voltammetric signal on a pyrolytic graphite electrode. The technique was tested under optimal conditions and various situations limiting or precluding the ligation reaction (such as DNA substrates lacking 5'-phosphate or containing a base mismatch at the nick junction, or application of incompatible cofactor), and utilized for the analysis of the nick-joining activity of a range of recombinant Escherichia coli DNA ligase constructs. The novel technique provides a fast, versatile, specific, and sensitive electrochemical assay of DNA ligase activity.

Evaluation of a magnetic particles-based chemiluminescence enzyme immunoassay for Golgi protein 73 in human serum.

Science.gov (United States)

Liu, Xiangyi; Wan, Xiaohua; Lu, Sheng; Zhang, Lijun; Yu, Shaohua; Lu, Xinxin

2015-05-20

Golgi protein 73 (GP73) is regarded as a potential serum biomarker for early diagnosis of hepatocellular carcinoma (HCC). We developed a rapid magnetic particles-based chemiluminescence enzyme immunoassay (MPs-CLEIA) for the determination of serum GP73. Fluorescein isothiocyanate (FITC) and alkaline phosphatase (ALP) were used to label 2 different monoclonal antibodies to GP73. Serum GP73 was captured with labeled antibodies and formed a sandwiched immunoreaction. The magnetic particles (MPs) coated with anti-FITC antibody were used as a means of separation of the GP73 protein from other serum proteins. After adding the enzyme substrate solution, the relative light unit (RLU) was measured. A MPs-CLEIA for serum GP73 was established and evaluated. The RLU was directly proportional to the concentration of GP73. The method linearity was 5-600 μg/l. Limit of the blank was 2.19 μg/l. The intra- and inter-assay imprecision was 73-0.89), and the sensitivity and specificity, with cut-off value of 115.6 μg/l, were 75.4% and 92.1%, respectively. The proposed method demonstrates an acceptable performance for quantifying serum GP73. This assay could be appropriate for routine use in clinical laboratories. Copyright © 2015 Elsevier B.V. All rights reserved.

Development of an immunomagnetic bead-based time-resolved fluorescence immunoassay for rapid determination of levels of carcinoembryonic antigen in human serum

Energy Technology Data Exchange (ETDEWEB)

Hou Jingyuan; Liu Tiancai; Lin Guanfeng; Li Zhixiong; Zou Liping; Li Ming [Institute of Antibody Engineering, School of Biotechnology, Southern Medical University, Guangzhou 510515 (China); Wu Yingsong, E-mail: wg@fimmu.com [Institute of Antibody Engineering, School of Biotechnology, Southern Medical University, Guangzhou 510515 (China)

2012-07-13

Highlights: Black-Right-Pointing-Pointer Magnetic beads was used as the solid phase for TRFIA. Black-Right-Pointing-Pointer The linearity range was broadened greatly compared with conventional TRFIA method. Black-Right-Pointing-Pointer The analysis time was significantly shorter compared with conventional TRFIA method. Black-Right-Pointing-Pointer This method could be developed for practical clinical detections of tumor-associated antigens. - Abstract: A novel immunoassay for the determination of tumor markers in human serum was established by combining a time-resolved fluoroimmunoassay (TRFIA) and immunomagnetic separation. Based on a sandwich-type immunoassay format, analytes in samples were captured by magnetic beads coated with one monoclonal antibody and 'sandwiched' by another monoclonal antibody labeled with europium chelates. The immunocomplex was separated and washed by exposure to a magnetic field and treatment with enhancement solution; fluorescence was then measured according to the number of europium ions dissociated. Levels of the model analyte, carcinoembryonic antigen (CEA), were determined in a linear range (1-1000 ng mL{sup -1}) with a limit of detection of 0.5 ng mL{sup -1} under optimal conditions. The reproducibility, recovery, and specificity of the immunoassay were demonstrated to be acceptable. To evaluate this novel assay for clinical applications, 239 serum samples were evaluated. Compared with the conventional TRFIA and chemiluminescence immunoassay (CLIA), the correlation coefficients of the developed immunoassay were 0.985 and 0.975, respectively. These results showed good correlation and confirmed that our method is feasible and could be used for the clinical determination of CEA (or other tumor antigens) in human serum.

Monoclonal antibody-based immunoassays.

Science.gov (United States)

Appleby, P; Reischl, U

1998-01-01

An immunoassay may be defined as an assay that employs an immunological reagent, usually an antibody, to confer specificity for the ligand being measured. As a corollary to this, the discovery, and subsequent development, of monoclonal antibodies (MAbs) has greatly expanded the application and use of immunoassays. Polyclonal reagents, with their associated problems of specificity and quality control, have now been largely replaced by readily available MAbs of potential immortality and well-defined specificity and affinity. This has resulted, in the last two decades, in a great expansion in the range of immunoassays available and also a significant improvement in their reproducibility and reliability.

Materials for Microfluidic Immunoassays: A Review.

Science.gov (United States)

Mou, Lei; Jiang, Xingyu

2017-08-01

Conventional immunoassays suffer from at least one of these following limitations: long processing time, high costs, poor user-friendliness, technical complexity, poor sensitivity and specificity. Microfluidics, a technology characterized by the engineered manipulation of fluids in channels with characteristic lengthscale of tens of micrometers, has shown considerable promise for improving immunoassays that could overcome these limitations in medical diagnostics and biology research. The combination of microfluidics and immunoassay can detect biomarkers with faster assay time, reduced volumes of reagents, lower power requirements, and higher levels of integration and automation compared to traditional approaches. This review focuses on the materials-related aspects of the recent advances in microfluidics-based immunoassays for point-of-care (POC) diagnostics of biomarkers. We compare the materials for microfluidic chips fabrication in five aspects: fabrication, integration, function, modification and cost, and describe their advantages and drawbacks. In addition, we review materials for modifying antibodies to improve the performance of the reaction of immunoassay. We also review the state of the art in microfluidic immunoassays POC platforms, from the laboratory to routine clinical practice, and also commercial products in the market. Finally, we discuss the current challenges and future developments in microfluidic immunoassays. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Detection of Avian Influenza Virus by Fluorescent DNA Barcode-based Immunoassay with Sensitivity Comparable to PCR

DEFF Research Database (Denmark)

Cao, Cuong; Dhumpa, Raghuram; Bang, Dang Duong

2010-01-01

involves the sandwiching of the target AIV between magnetic immunoprobes and barcode-carrying immunoprobes. Because each barcode-carrying immunoprobe is functionalized with a multitude of fluorophore-DNA barcode strands, many DNA barcodes are released for each positive binding event resulting......In this paper, a coupling of fluorophore-DNA barcode and bead-based immunoassay for detecting avian influenza virus (AIV) with PCR-like sensitivity is reported. The assay is based on the use of sandwich immunoassay and fluorophore-tagged oligonucleotides as representative barcodes. The detection...

Advanced techniques in immunoassay

International Nuclear Information System (INIS)

Toth, G.

1982-01-01

A brief overview of the development history of radioimmunoassay and related techniques with their theory and practice are given. A comparison of radioimmunoassay (RIA), enzyme immunoassay (EIA), spin immunoassay (SIA), sequential saturation analysis (SSA) etc., based on their main parameters, and their fields of application and recent trends are presented. (Sz.J.)

Radiolabelling for immunoassay

International Nuclear Information System (INIS)

Chapman, R.S.

1998-01-01

Since the early 1960s labelled compounds employed in immunoassay techniques, both radioimmunoassay and immunoradiometric assay, have involved radioisotopes typically 3 H (tritium) and 125 Iodine. With the advent of increasingly stringent governmental regulations regarding usage and disposal of radioisotopes and the impetus of research towards improved immunoassay sensitivity following the discovery of monoclonal antibodies and their application to excess reagent immunometric assay methodology, radioisotopic labels are gradually being replaced by non-isotopic labels: enzyme, fluorescence and chemiluminescence

Integration of microcoils for on-chip immunosensors based on magnetic nanoparticles capture

Directory of Open Access Journals (Sweden)

Olivier Lefebvre

2017-04-01

Full Text Available Immunoassays using magnetic nanoparticles (MNP are generally performed under the control of permanent magnet close to the micro-tube of reaction. Using a magnet gives a powerful method for driving MNP but remains unreliable or insufficient for a fully integrated immunoassay on lab-on-chip. The aim of this study is to develop a novel lab-on-chip concept for high efficient immunoassays to detect ovalbumin (Biodefense model molecule with microcoils employed for trapping MNP during the biofunctionalization steps. The objectives are essentially to optimize their efficiency for biological recognition by assuring a better bioactivity (antibodies-ovalbumin, and detect small concentrations of the targeted protein (~10 pg/mL. In this work, we studied the response of immunoassays complex function of ovalbumin concentration. The impact of MNP diameter in the biografting protocol was studied and permitted to choose a convenient MNP size for efficient biorecognition. We realized different immunoassays by controlling MNP in test tube and in microfluidic device using a permanent magnet. The comparison between these two experiments allows us to highlight an improvement of the limit of detection in microfluidic conditions by controlling MNP trapping with a magnet. Keywords: Bacteria, Lab-on-chip, ELISA, Magnetic nanoparticles, Ovalbumin, Microcoils, Fluorescent microscopy

Survey of immunoassay techniques for biological analysis

International Nuclear Information System (INIS)

Burtis, C.A.

1986-10-01

Immunoassay is a very specific, sensitive, and widely applicable analytical technique. Recent advances in genetic engineering have led to the development of monoclonal antibodies which further improves the specificity of immunoassays. Originally, radioisotopes were used to label the antigens and antibodies used in immunoassays. However, in the last decade, numerous types of immunoassays have been developed which utilize enzymes and fluorescent dyes as labels. Given the technical, safety, health, and disposal problems associated with using radioisotopes, immunoassays that utilize the enzyme and fluorescent labels are rapidly replacing those using radioisotope labels. These newer techniques are as sensitive, are easily automated, have stable reagents, and do not have a disposal problem. 6 refs., 1 fig., 2 tabs

Sandwich immunoassay for alpha-fetoprotein in human sera using gold nanoparticle and magnetic bead labels along with resonance Rayleigh scattering readout

International Nuclear Information System (INIS)

Lu, Yao; Huang, Xiangyi; Ren, Jicun

2013-01-01

We describe a sensitive sandwich immunoassay for alpha-fetoprotein (AFP). It is making use of gold nanoparticles (GNPs) and magnetic beads (MBs) as labels, and of resonance Rayleigh scattering for detection. Two antibodies were labeled with GNPs and MBs, respectively, and MB-antigen-GNP complexes were formed in the presence of antigens. The MB labels also serve as solid phase carriers that can be used to magnetically separate the immuno complex. The GNP labels are used as optical probes, and Rayleigh scattering was used to determine the concentration of free GNPs-antibody after separation of the MB-antigen-GNP complexes. The concentration of AFP is related to the intensity of light scattered by free GNPs in the 13.6 pM to 436 pM concentration range, and the limit of detection is 13.6 pM. The method was applied to the determination of AFP in sera of cancer patients, and the results agree well with those obtained by conventional ELISA. (author)

Floating Chip Mounting System Driven by Repulsive Force of Permanent Magnets for Multiple On-Site SPR Immunoassay Measurements

Science.gov (United States)

Horiuchi, Tsutomu; Tobita, Tatsuya; Miura, Toru; Iwasaki, Yuzuru; Seyama, Michiko; Inoue, Suzuyo; Takahashi, Jun-ichi; Haga, Tsuneyuki; Tamechika, Emi

2012-01-01

We have developed a measurement chip installation/removal mechanism for a surface plasmon resonance (SPR) immunoassay analysis instrument designed for frequent testing, which requires a rapid and easy technique for changing chips. The key components of the mechanism are refractive index matching gel coated on the rear of the SPR chip and a float that presses the chip down. The refractive index matching gel made it possible to optically couple the chip and the prism of the SPR instrument easily via elastic deformation with no air bubbles. The float has an autonomous attitude control function that keeps the chip parallel in relation to the SPR instrument by employing the repulsive force of permanent magnets between the float and a float guide located in the SPR instrument. This function is realized by balancing the upward elastic force of the gel and the downward force of the float, which experiences a leveling force from the float guide. This system makes it possible to start an SPR measurement immediately after chip installation and to remove the chip immediately after the measurement with a simple and easy method that does not require any fine adjustment. Our sensor chip, which we installed using this mounting system, successfully performed an immunoassay measurement on a model antigen (spiked human-IgG) in a model real sample (non-homogenized milk) that included many kinds of interfering foreign substances without any sample pre-treatment. The ease of the chip installation/removal operation and simple measurement procedure are suitable for frequent on-site agricultural, environmental and medical testing. PMID:23202030

Horseradish peroxidase and antibody labeled gold nanoparticle probe for amplified immunoassay of ciguatoxin in fish samples based on capillary electrophoresis with electrochemical detection.

Science.gov (United States)

Zhang, Zhaoxiang; Liu, Ying; Zhang, Chaoying; Luan, Wenxiu

2015-03-01

This paper describes a new amplified immunoassay with horseradish peroxidase (HRP) and antibody (Ab) labeled gold nanoparticles (AuNPs) probe hyphenated to capillary electrophoresis (CE) with electrochemical (EC) detection for ultrasensitive determination of ciguatoxin CTX1B. AuNPs were conjugated with HRP and Ab, and then incubated with limited amount of CTX1B to produce immunocomplex. The immunoreactive sample was injected into capillary for CE separation and EC detection. Enhanced sensitivity was obtained by adopting the AuNPs as carriers of HRP and Ab at high HRP/Ab molar ratio. The calibration curve of CTX1B was in the range of 0.06-90 ng/mL. The detection limit was 0.045 ng/mL, which is 38-fold lower than that of HPLC-MS method for CTX1B analysis. The proposed method was successfully applied for the quantification of CTX1B in contamined fish samples by simultaneously labeling Ab and HRP on AuNPs. The amplified IA with HRP and Ab labeled AuNPs probe hyphenated to CE and EC detection provides a sensitive analytical approach for the determination of trace ciguatoxin in complex samples. Copyright © 2015 Elsevier Ltd. All rights reserved.

Electrochemical fabrication, microstructure and magnetic properties of Sm{sub 2}Co{sub 17}/Fe{sub 7}Co{sub 3} dual phase nanocomposite

Energy Technology Data Exchange (ETDEWEB)

Cui, Chunxiang, E-mail: hutcui@hebut.edu.cn [Key Lab. for New Type of Functional Materials in Hebei Province, Hebei University of Technology, No.8, Road No.1, Dingzigu, Hongqiao District, Tianjin 300130 (China); Chen, Fenghua [Tianjin Sanhuan Lucky New Materials Inc., Tianjin Economical-Technological Development Area (TEDA), Tianjin 300457 (China); Yang, Wei; Li, Hongfang; Liu, Qiaozhi; Sun, Jibing [Key Lab. for New Type of Functional Materials in Hebei Province, Hebei University of Technology, No.8, Road No.1, Dingzigu, Hongqiao District, Tianjin 300130 (China)

2015-06-15

By utilizing alternate electrochemical reaction, atomic migration and deposition of Fe, Co, Sm and other chemical substances in the electrochemical solution, a large number of Sm{sub 2}Co{sub 17}/Fe{sub 7}Co{sub 3} dual phase nanowire arrays were carried out in the anodic aluminum oxide (AAO) template with highly uniform and orderly. The Sm{sub 2}Co{sub 17}/Fe{sub 7}Co{sub 3} dual phase nanowire arrays with diameter of 50 nm and length of 12 μm have the smooth surface and uniform diameter. The morphology and microstructure of annealed Sm{sub 2}Co{sub 17}/Fe{sub 7}Co{sub 3} dual phase nanowires were observed and analyzed using SEM, TEM and HRTEM. Compared with single-phase nanowires, dual phase magnetic nanowires have higher coercivity and saturation magnetization. In this composite system, both the hard and the soft phases have a high Curie temperature, therefore, we believe that the Sm{sub 2}Co{sub 17}/Fe{sub 7}Co{sub 3} dual phase nanowire arrays is a new type of high-temperature magnetic composites. - Highlights: • Sm{sub 2}Co{sub 17}/Fe{sub 7}Co{sub 3} dual phase nanowires were prepared by electrochemical method. • The interface pinning is the main factor to improve anisotropy field of the nanowires. • The dual phase magnetic nanowires have higher coercivity and saturation magnetization.

Rapid determination of recent cocaine use with magnetic particles-based enzyme immunoassays in serum, saliva, and urine fluids.

Science.gov (United States)

Vidal, Juan C; Bertolín, Juan R; Bonel, Laura; Asturias, Laura; Arcos-Martínez, M Julia; Castillo, Juan R

2016-06-05

Cocaine is one of the most worldwide used illicit drugs. We report a magnetic particles-based enzyme-linked immunoassay (mpEIA) method for the rapid and sensitive determination of cocaine (COC) in saliva, urine and serum samples. Under optimized conditions, the limits of detections were 0.09ngmL(-1) (urine), 0.15ngmL(-1) (saliva), and 0.06ngmL(-1) COC (human serum). Sensitivities were in the range EC50=0.6-2.5ngmL(-1) COC. The cross-reactivity with the principal metabolite benzoylecgonine (BZE) was only 1.6%. Recovering percentages of doped samples (0, 10, 50, and 100ngmL(-1) of COC) ranged from about 86-111%. Some advantages of the developed mpEIA over conventional ELISA kits are faster incubations, improved reproducibility, and consumption of lower amounts of antibody and enzyme conjugates due to the use of magnetic beads. The reported method was validated following the guidelines on bioanalytical methods of the European Medicines Agency (2011). Unmetabolized COC detection has a great interest in pharmacological, pharmacokinetics, and toxicokinetics studies, and can be used to detect a very recent COC use (1-6h). Copyright © 2016 Elsevier B.V. All rights reserved.

New advances in electrochemical biosensors for the detection of toxins: Nanomaterials, magnetic beads and microfluidics systems. A review

International Nuclear Information System (INIS)

Reverté, Laia; Prieto-Simón, Beatriz; Campàs, Mònica

2016-01-01

The use of nanotechnology in bioanalytical devices has special advantages in the detection of toxins of interest in food safety and environmental applications. The low levels to be detected and the small size of toxins justify the increasing number of publications dealing with electrochemical biosensors, due to their high sensitivity and design versatility. The incorporation of nanomaterials in their development has been exploited to further increase their sensitivity, providing simple and fast devices, with multiplexed capabilities. This paper gives an overview of the electrochemical biosensors that have incorporated carbon and metal nanomaterials in their configurations for the detection of toxins. Biosensing systems based on magnetic beads or integrated into microfluidics systems have also been considered because of their contribution to the development of compact analytical devices. The roles of these materials, the methods used for their incorporation in the biosensor configurations as well as the advantages they provide to the analyses are summarised. - Highlights: • Nanomaterials improve the performance of electrochemical biosensors. • Carbon nanomaterials can act as electrocatalysts or label supports in biosensors. • Metal nanomaterials can act as nanostructured supports or labels in biosensors. • Magnetic beads are exploited as immobilisation supports and/or label carriers.

New advances in electrochemical biosensors for the detection of toxins: Nanomaterials, magnetic beads and microfluidics systems. A review

Energy Technology Data Exchange (ETDEWEB)

Reverté, Laia [IRTA, Carretera Poble Nou km. 5.5, 43540 Sant Carles de la Ràpita, Tarragona (Spain); Prieto-Simón, Beatriz [ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, Future Industries Institute, University of South Australia, SA 5095 (Australia); Campàs, Mònica, E-mail: monica.campas@irta.cat [IRTA, Carretera Poble Nou km. 5.5, 43540 Sant Carles de la Ràpita, Tarragona (Spain)

2016-02-18

The use of nanotechnology in bioanalytical devices has special advantages in the detection of toxins of interest in food safety and environmental applications. The low levels to be detected and the small size of toxins justify the increasing number of publications dealing with electrochemical biosensors, due to their high sensitivity and design versatility. The incorporation of nanomaterials in their development has been exploited to further increase their sensitivity, providing simple and fast devices, with multiplexed capabilities. This paper gives an overview of the electrochemical biosensors that have incorporated carbon and metal nanomaterials in their configurations for the detection of toxins. Biosensing systems based on magnetic beads or integrated into microfluidics systems have also been considered because of their contribution to the development of compact analytical devices. The roles of these materials, the methods used for their incorporation in the biosensor configurations as well as the advantages they provide to the analyses are summarised. - Highlights: • Nanomaterials improve the performance of electrochemical biosensors. • Carbon nanomaterials can act as electrocatalysts or label supports in biosensors. • Metal nanomaterials can act as nanostructured supports or labels in biosensors. • Magnetic beads are exploited as immobilisation supports and/or label carriers.

Effects of size reduction on the structure and magnetic properties of core-shell Ni3Si/silica nanoparticles prepared by electrochemical synthesis

Czech Academy of Sciences Publication Activity Database

Pigozzi, G.; Mukherji, D.; Elerman, Y.; Strunz, Pavel; Gilles, R.; Hoelzel, M.; Barbier, B.; Schmutz, P.

2014-01-01

Roč. 584, JAN (2014), s. 119-127 ISSN 0925-8388 Institutional support: RVO:61389005 Keywords : intermetallics * nanostructured materials * transition metal alloys and compounds * electrochemical synthesis * crystal structure * magnetic measurements Subject RIV: BM - Solid Matter Physics ; Magnetism Impact factor: 2.999, year: 2014

Hydrogel nanoparticle based immunoassay

Science.gov (United States)

Liotta, Lance A; Luchini, Alessandra; Petricoin, Emanuel F; Espina, Virginia

2015-04-21

An immunoassay device incorporating porous polymeric capture nanoparticles within either the sample collection vessel or pre-impregnated into a porous substratum within fluid flow path of the analytical device is presented. This incorporation of capture particles within the immunoassay device improves sensitivity while removing the requirement for pre-processing of samples prior to loading the immunoassay device. A preferred embodiment is coreshell bait containing capture nanoparticles which perform three functions in one step, in solution: a) molecular size sieving, b) target analyte sequestration and concentration, and c) protection from degradation. The polymeric matrix of the capture particles may be made of co-polymeric materials having a structural monomer and an affinity monomer, the affinity monomer having properties that attract the analyte to the capture particle. This device is useful for point of care diagnostic assays for biomedical applications and as field deployable assays for environmental, pathogen and chemical or biological threat identification.

Establishment of a novel immunoassay system for rapid detection of 2,4-dichlorophenoxyacetic acid residues based on magnetic-fluorescent probes

Directory of Open Access Journals (Sweden)

WANG Yuanfeng

2014-12-01

Full Text Available A novel immunoassay system based on magnetic-fluorescent probes was established to detect 2.4-dichlorophenoxyacetic acid (2,4-D residue in liquid system in food and agricultural products.The composites of anti-2,4-D antibody bound to Fe3O4@SiO2-NH2 was employed as the solid phase as well as magnetic probe.The composites composed of 2,4-D-OVA labeled with CdTe@SiO2-NH2 as the fluorescent probe was used to produce fluorescent signal.2,4-D and its fluorescent probe competed binding the antibody on the surface of the magnetic probe.The optimization of 2,4-D-OVA dosage,coupling PH and reaction time in preparing the fluorescent probe were investigated.It showed that in the synthesis of fluorescent probe 8.2 was the optimal pH,70 min was the optimal coupling time,500 μL amount of 2,4-D-OVA.The standard curve was obtained with the concentration of 2,4-D and the maximum fluorescence intensity.The detection limit of the assay was gotten and it was 3.55×10-8.One reaction step and one washing step were needed.The assay significantly shortened the testing time and amplified the detection signal compared with classic ELISA.

Magnetophoretic potential at the movement of cluster products of electrochemical reactions in an inhomogeneous magnetic field

International Nuclear Information System (INIS)

Gorobets, O. Yu.; Gorobets, Yu. I.; Rospotniuk, V. P.

2015-01-01

An electric field arises from the influence of a nonuniform static magnetic field on charged colloid particles with magnetic susceptibility different from that of the surrounding liquid. It arises, for example, under the influence of a nonuniform static magnetic field in clusters of electrochemical reaction products created during metal etching, deposition, and corrosion processes without an external electric current passing through an electrolyte near a magnetized electrode surface. The corresponding potential consists of a Nernst potential of inhomogeneous distribution of concentration of colloid particles and a magnetophoretic potential (MPP). This potential has been calculated using a thermodynamic approach based on the equations of thermodynamics of nonequilibrium systems and the Onsager relations for a mass flow of correlated magnetic clusters under a gradient magnetic force in the electrolyte. The conditions under which the MPP contribution to the total electric potential may be significant are discussed with a reference to the example of a corroding spherical ferromagnetic steel electrode

Magnetophoretic potential at the movement of cluster products of electrochemical reactions in an inhomogeneous magnetic field

Energy Technology Data Exchange (ETDEWEB)

Gorobets, O. Yu., E-mail: pitbm@ukr.net; Gorobets, Yu. I., E-mail: Gorobets@imag.kiev.ua [National Technical University of Ukraine “KPI”, Peremogy Avenue 37, Kyiv 03056 (Ukraine); Institute of Magnetism NAS of Ukraine and National Academy of Sciences of Ukraine, Vernadsky Avenue, 36-b, Kyiv 03142 (Ukraine); Rospotniuk, V. P. [National Technical University of Ukraine “KPI”, Peremogy Avenue 37, Kyiv 03056 (Ukraine)

2015-08-21

An electric field arises from the influence of a nonuniform static magnetic field on charged colloid particles with magnetic susceptibility different from that of the surrounding liquid. It arises, for example, under the influence of a nonuniform static magnetic field in clusters of electrochemical reaction products created during metal etching, deposition, and corrosion processes without an external electric current passing through an electrolyte near a magnetized electrode surface. The corresponding potential consists of a Nernst potential of inhomogeneous distribution of concentration of colloid particles and a magnetophoretic potential (MPP). This potential has been calculated using a thermodynamic approach based on the equations of thermodynamics of nonequilibrium systems and the Onsager relations for a mass flow of correlated magnetic clusters under a gradient magnetic force in the electrolyte. The conditions under which the MPP contribution to the total electric potential may be significant are discussed with a reference to the example of a corroding spherical ferromagnetic steel electrode.

The right environment for the immunoassay

International Nuclear Information System (INIS)

Emon, J.M. Van; Gerlach, C.L.

1995-01-01

For the US Environmental Protection Agency (EPA), the first in-house research effort began in 1987, when results of an early immunoassay field study verified the technology's potential for environmental applications. Looking at the fundamental features of immunochemical reactions from the clinical laboratories, analytical chemists realized the potential value of these methods for hazardous waste site characterization and pesticide monitoring. Immunoassays rely on the interaction between an antibody and a target analyte. For environmental purposes, enzyme immunoassays are generally used. After the target analyte binds to the antibody, an enzymatic reaction yields a colorimetric change. This change, read visually or by a spectrophotometer, indicates the concentration of the target analyte. Promising results with assays for compounds (such as paraquat and pentachlorophenol) and compound groups (such as total petroleum hydrocarbons and polychlorinated biphenyls) spurred interest among various entrepreneurs. The first target market for immunoassays was environmental engineers and field crews who needed quick answers on-site to determine the direction of further remediation efforts

Preparation of RGO/Fe{sub 3}O{sub 4}/poly (acrylic acid) hydrogel nanocomposites with improved magnetic, thermal and electrochemical properties

Energy Technology Data Exchange (ETDEWEB)

Didehban, K.H., E-mail: Didehban95@gmail.com; Mohammadi, L.; Azimvand, J.

2017-07-01

A hydrogel nanocomposite composed of reduced graphene oxide (RGO), iron oxide (Fe{sub 3}O{sub 4}) nanoparticles, and polyacrylic acid (PAA) was prepared using radical polymerization. Different percentages of RGO, Fe{sub 3}O{sub 4}, and PAA were used to prepare the nanocomposite. Fourier transform infrared spectroscopy (FTIR) results confirmed the formation of the nanocomposite’s chemical structure. X-ray power diffraction (XRD) patterns revealed the principal peak’s 2θ value to be 77.39° with the size of the nanocomposite particles estimated at 96 nm. Results indicated that the electrochemical capacity of the nanocomposites was controlled by the weight percentage of RGO. Increases to the potential scan rate reduced porosity and surface area, thereby decreasing the electrochemical capacity of the nanocomposites. Moreover, increasing the percentage of Fe{sub 3}O{sub 4} nanoparticles in the nanocomposites improved their magnetic characteristics and thermal properties. The latter also improved when the RGO percentage increased. - Highlights: • A hydrogel nanocomposite composed of RGO/Fe{sub 3}O{sub 4}/PAA was synthesized successfully. • Increasing the percentage of iron nanoparticles improved magnetic properties. • Increasing the percentage of RGO improved thermal and electrochemical capacity. • The Fe{sub 3}O{sub 4} nanoparticles directly affected magnetic properties.

A magnetic particle micro-trap for large trapping surfaces

KAUST Repository

Gooneratne, Chinthaka P.

2012-01-08

Manipulation of micron-size magnetic particles of the superparamagnetic type contributes significantly in many applications like controlling the antibody/antigen binding process in immunoassays. Specifically, more target biomolecules can be attached/tagged and analyzed since the three dimensional structure of the magnetic particles increases the surface to volume ratio. Additionally, such biomolecular-tagged magnetic particles can be easily manipulated by an external magnetic field due to their superparamagnetic behavior. Therefore, magnetic particle- based immunoassays are extensively applied in micro-flow cytometry. The design of a square-loop micro-trap as a magnetic particle manipulator as well as numerical and experimental analysis is presented. Experimental results showed that the micro-trap could successfully trap and concentrate magnetic particles from a large to a small area with a high spatial range.

A magnetic particle micro-trap for large trapping surfaces

KAUST Repository

Gooneratne, Chinthaka P.; Liang, Cai; Giouroudi, Ioanna; Kosel, Jü rgen

2012-01-01

Manipulation of micron-size magnetic particles of the superparamagnetic type contributes significantly in many applications like controlling the antibody/antigen binding process in immunoassays. Specifically, more target biomolecules can be attached/tagged and analyzed since the three dimensional structure of the magnetic particles increases the surface to volume ratio. Additionally, such biomolecular-tagged magnetic particles can be easily manipulated by an external magnetic field due to their superparamagnetic behavior. Therefore, magnetic particle- based immunoassays are extensively applied in micro-flow cytometry. The design of a square-loop micro-trap as a magnetic particle manipulator as well as numerical and experimental analysis is presented. Experimental results showed that the micro-trap could successfully trap and concentrate magnetic particles from a large to a small area with a high spatial range.

Rapid micromotor-based naked-eye immunoassay.

Science.gov (United States)

de Ávila, Berta Esteban-Fernández; Zhao, Mingjiao; Campuzano, Susana; Ricci, Francesco; Pingarrón, José M; Mascini, Marcello; Wang, Joseph

2017-05-15

A dynamic micromotor-based immunoassay, exemplified by cortisol detection, based on the use of tubular micromotors functionalized with a specific antibody is described. The use of antibody-functionalized micromotors offers huge acceleration of both direct and competitive cortisol immunoassays, along with greatly enhanced sensitivity of direct and competitive immunoassays. The dramatically improved speed and sensitivity reflect the greatly increased likelihood of antibody-cortisol contacts and fluid mixing associated with the dynamic movement of these microtube motors and corresponding bubble generation that lead to a highly efficient and rapid recognition process. Rapid naked-eye detection of cortisol in the sample is achieved in connection to use of horseradish peroxidase (HRP) tag and TMB/H 2 O 2 system. Key parameters of the competitive immunoassay (e.g., incubation time and reaction volume) were optimized. This fast visual micromotor-based sensing approach enables "on the move" specific detection of the target cortisol down to 0.1μgmL -1 in just 2min, using ultrasmall (50µL) sample volumes. Copyright © 2017 Elsevier B.V. All rights reserved.

Sensitive electrochemical determination of α-fetoprotein using a glassy carbon electrode modified with in-situ grown gold nanoparticles, graphene oxide and MWCNTs acting as signal amplifiers

International Nuclear Information System (INIS)

Gao, Yan-Sha; Zhu, Xiao-Fei; Yang, Tao-Tao; Xu, Jing-Kun; Zhang, Kai-Xin; Lu, Li-Min

2015-01-01

The authors describe an electrochemical immunoassay for α-fetoprotein (α-FP) using a glassy carbon electrode (GCE) modified with a nanocomposite made from gold nanoparticles, graphene oxide and multi-walled carbon nanotubes (AuNPs/GO-MWCNTs) and acting as a signal amplification matrix. The nanocomposite was synthesized in a one-pot redox reaction between GO and HAuCl 4 without using an additional reductant. The stepwise assembly of the immunoelectrode was characterized by means of cyclic voltammetry and electrochemical impedance spectroscopy. The interaction of antigen and antibody on the surface of the electrode creates a barrier for electrons and causes retarded electron transfer, this resulting in decreased signals in differential pulse voltammetry of hexacyanoferrate which is added as an electrochemical probe. Using this strategy and by working at a potential of 0.2 V (vs. SCE), a wide analytical range (0.01 - 100 ng∙mL -1 ) is covered. The correlation coefficient is 0.9929, and the limit of detection is as low as 3 pg∙mL -1 at a signal-to-noise ratio of 3. This electrochemical immunoassay combines the specificity of an immunological detection scheme with the sensitivity of an electrode modified with AuNPs and GO-MWCNTs. (author)

Effects of magnetic processing on electrochemical and photoelectrochemical properties of electrodes modified with nanoclusters of a phenothiazine-C60 linked compound

International Nuclear Information System (INIS)

Yonemura, H; Yamada, S; Wakita, Y; Moribe, S; Fujiwara, Y; Tanimoto, Y

2009-01-01

Effects of magnetic processing on morphological, electrochemical, and photoelectrochemical properties of electrodes modified with nanoclusters of a phenothiazine-C 60 linked compound with four methylene group (Ph(4)C 60 ) were examined in the absence and presence of magnetic processing with three different magnetic environments due to strong magnetic field. The AFM measurements indicated that the morphologies of nanostructures of Ph(4)C 60 varied with magnetic enviroments as comparison with that in the absence of magnetic processing. At top position (5.6 T; - 940 T 2 /m) with hypogravity, large spherical nanoclusters (60∼70 nm diameter) were observed as comparion with those (ca. 20 nm diameter) in the absence of magnetic processing. At middle positon (15 T; 0 T 2 /m) with normal gravity, the fiber-like nanostructure was observed. At bottom position (9.8 T; + 1070T 2 /m) with hypergravity, the rod-like nanostrucure was observed. The interesting results might be ascribed to the different solvent properties due to the different rates of evaporation of two solvents in the toluene-acetonitrile mixed solvent during drying process under various magnetic environments. First reduction peaks due to C 60 moiety of Ph(4)C 60 nanostrucures in the presence of magnetic processing at three different positions were negative-shifted as comparison with that in the absence of magnetic processing. Potential dependencies of the photocurrents of the electrodes modified with Ph(4)C 60 nanostrucures in the presence of magnetic processing at three positions were also different from that in the absence of magnetic processing. The magnetic field effects in AFM, and electrochemical and photoelectrochemical measurements are most likely ascribed to the difference of the reduction potentials due to C 60 clusters between the absence and presence of magnetic processing due to the morphological change of Ph(4)C 60 nanostrucures.

Electrochemical sensors in breast cancer diagnostics and follow-up

Directory of Open Access Journals (Sweden)

Raquel Marques

2015-12-01

Full Text Available Purpose: The detection of tumor biomarkers can have a major contribution to the management of breast cancer. So far the only serum biomarker in current use in breast cancer is the cancer antigen 15-3 (CA15-3. This biomarker is used in advanced breast cancer to monitor patients and to help to identify treatment failure. The human epidermal growth factor receptor 2 (HER 2 is another biomarker whose characterization is usually made in tissue samples from primary tumour or metastasis and has been used as a prognostic factor but mainly as a target in immunotherapy treatment. Some previous studies suggest that the detection of the extracellular domain of HER2 (HER2-ECD in blood can be a prognostic factor, with even better results than its detection in tissue. Recent techniques for circulating protein biomarker detection use immunoassays, but some are, for example, not sufficiently sensitive for the detection of low biomarker concentrations. To overcome some of these problems, electrochemical (biosensors, and especially the ones using voltammetric detection, can be adequate alternatives because of their high selectivity and sensitivity which allows early detection of many diseases. Furthermore, electrochemical (biosensors are excellent to be included into point-of-care devices due to their fast response, simplicity, low cost, easy miniaturization and integration into automatic systems. Another advantage is the possibility of combining individual sensors into multiplexed detection systems. Like this they can provide fast recording of biomarker profiles of tumours which can play an important role in early detection and personalized medicine.Methods: Both individual as well as multiplexed electrochemical immunosensors were developed for the detection of CA15-3 and HER2-ECD. For this purpose a sandwich immunoassay was employed and the analytical signal was based on the voltammetric detection of enzymatically deposited silver. Screen-printed carbon

Electrochemical alloying of immiscible Ag and Co for their structural and magnetic analyses

Energy Technology Data Exchange (ETDEWEB)

Santhi, Kalavathy [Material Science Centre, Department of Nuclear Physics, University of Madras, Guindy Campus, Chennai 600025 (India); Department of Physics, Women’s Christian College, Chennai 600006 (India); Kumarsan, Dhanapal [Material Science Centre, Department of Nuclear Physics, University of Madras, Guindy Campus, Chennai 600025 (India); Vengidusamy, Naryanan [Department of Inorganic Chemistry, University of Madras, Guindy Campus, Chennai 600025 (India); Arumainathan, Stephen, E-mail: stephen_arum@hotmail.com [Material Science Centre, Department of Nuclear Physics, University of Madras, Guindy Campus, Chennai 600025 (India)

2017-07-01

Highlights: • Ag-Co alloy has been prepared using pulsed electrodeposition method. • Wide range of Ag composition in the alloy was obtained. • XPS measurement evident the Ag and Co in metallic nature. • The electrodeposition method develop dendrite like morphology. • Detailed analysis of magnetic behaviour is carried out. - Abstract: Electrochemical alloying of immiscible Ag and Co was carried out at different current densities from electrolytes of two different concentrations, after optimizing the electrolytic bath and operating conditions. The samples obtained were characterized using X-ray diffraction to confirm the simultaneous deposition of Ag and Co and to determine their crystallographic structure. The atomic percentage of Ag and Co contents in the granular alloy was determined by ICP-OES analysis. The XPS spectra were observed to confirm the presence of Ag and Co in the metallic form in the granular alloy samples. The micrographs observed using scanning and transmission electron microscopes threw light on the surface morphology and the size of the particles. The magnetic nature of the samples was analyzed at room temperature by a vibration sample magnetometer. Their magnetic phase transition while heating was also studied to provide further evidence for the magnetic behaviour and the structure of the deposits.

Electrochemical alloying of immiscible Ag and Co for their structural and magnetic analyses

International Nuclear Information System (INIS)

Santhi, Kalavathy; Kumarsan, Dhanapal; Vengidusamy, Naryanan; Arumainathan, Stephen

2017-01-01

Highlights: • Ag-Co alloy has been prepared using pulsed electrodeposition method. • Wide range of Ag composition in the alloy was obtained. • XPS measurement evident the Ag and Co in metallic nature. • The electrodeposition method develop dendrite like morphology. • Detailed analysis of magnetic behaviour is carried out. - Abstract: Electrochemical alloying of immiscible Ag and Co was carried out at different current densities from electrolytes of two different concentrations, after optimizing the electrolytic bath and operating conditions. The samples obtained were characterized using X-ray diffraction to confirm the simultaneous deposition of Ag and Co and to determine their crystallographic structure. The atomic percentage of Ag and Co contents in the granular alloy was determined by ICP-OES analysis. The XPS spectra were observed to confirm the presence of Ag and Co in the metallic form in the granular alloy samples. The micrographs observed using scanning and transmission electron microscopes threw light on the surface morphology and the size of the particles. The magnetic nature of the samples was analyzed at room temperature by a vibration sample magnetometer. Their magnetic phase transition while heating was also studied to provide further evidence for the magnetic behaviour and the structure of the deposits.

Fabrication of highly catalytic silver nanoclusters/graphene oxide nanocomposite as nanotag for sensitive electrochemical immunoassay

Energy Technology Data Exchange (ETDEWEB)

Wang, Jiamian; Wang, Xiuyun; Wu, Shuo, E-mail: wushuo@dlut.edu.cn; Song, Jie; Zhao, Yanqiu; Ge, Yanqiu; Meng, Changgong

2016-02-04

Silver nanoclusters and graphene oxide nanocomposite (AgNCs/GRO) is synthesized and functionalized with detection antibody for highly sensitive electrochemical sensing of carcinoembryonic antigen (CEA), a model tumor marker involved in many cancers. AgNCs with large surface area and abundant amount of low-coordinated sites are synthesized with DNA as template and exhibit high catalytic activity towards the electrochemical reduction of H{sub 2}O{sub 2}. GRO is employed to assemble with AgNCs because it has large specific surface area, super electronic conductivity and strong π-π stacking interaction with the hydrophobic bases of DNA, which can further improve the catalytic ability of the AgNCs. Using AgNCs/GRO as signal amplification tag, an enzyme-free electrochemical immunosensing protocol is designed for the highly sensitive detection of CEA on the capture antibody functionalized immunosensing interface. Under optimal conditions, the designed immunosensor exhibits a wide linear range from 0.1 pg mL{sup −1} to 100 ng mL{sup −1} and a low limit of detection of 0.037 pg mL{sup −1}. Practical sample analysis reveals the sensor has good accuracy and reproducibility, indicating the great application prospective of the AgNCs/GRO in fabricating highly sensitive immunosensors, which can be extended to the detection of various kinds of low abundance disease related proteins. - Highlights: • An enzyme-free electrochemical immunosensor is reported for detecting proteins. • A silver nanocluster/graphene oxide composite is synthesized as nanotag. • The nanotags exhibit highly catalytic activity to the electro-reduction of H{sub 2}O{sub 2}. • The as-fabricated immunosensor could detect protein in serum samples.

Can LC and LC-MS ever replace immunoassays?

Directory of Open Access Journals (Sweden)

Timothy G. Cross

2016-10-01

Full Text Available Immunoassays have been the technology of choice for the analysis of biomolecules for many decades across a wide range of applications in research, diagnostics and infectious disease monitoring. There are good reasons for the wide adoption of immunoassays but even such a well established and characterised technique has limitations and as such investigators are looking at alternative technologies. One such alternative is liquid chromatography (LC and, more specifically, liquid chromatography coupled with mass spectrometry (LC-MS. This article will review both immunoassay and LC and LC-MS technologies and methodologies and discuss the advantages and limitations of both approaches. In addition, the next developments that will need to occur before there is widespread adoption of LC and LC-MS technology preferentially over immunoassays will be examined.

Determination of avian influenza A (H9N2) virions by inductively coupled plasma mass spectrometry based magnetic immunoassay with gold nanoparticles labeling

Science.gov (United States)

Xiao, Guangyang; Chen, Beibei; He, Man; Shi, Kaiwen; Zhang, Xing; Li, Xiaoting; Wu, Qiumei; Pang, Daiwen; Hu, Bin

2017-12-01

Avian influenza viruses are the pathogens of global poultry epidemics, and may even cause the human infections. Here, we proposed a novel inductively coupled plasma mass spectrometry (ICP-MS) based immunoassay with gold nanoparticles (Au NPs) labeling for the determination of H9N2 virions. Magnetic-beads modified with anti-influenza A H9N2 hemagglutinin mono-antibody (mAb-HA) were utilized for the capture of H9N2 virions in complex matrix; and Au NPs conjugated with mAb-HA were employed for the specific labeling of H9N2 virions for subsequent ICP-MS detection. With a sandwich immunoassay strategy, this method exhibited a high specificity for H9N2 among other influenza A virions such as H1N1 and H3N2. Under the optimized conditions, this method could detect as low as 0.63 ng mL- 1 H9N2 virions with the linear range of 2-400 ng mL- 1, the relative standard deviation for seven replicate detections of H9N2 virions was 7.2% (c = 10 ng mL- 1). The developed method was applied for the detection of H9N2 virions in real-world chicken dung samples, and the recovery for the spiking samples was 91.4-116.9%. This method is simple, rapid, sensitive, selective, reliable and has a good application potential for virions detection in real-world samples.

Status of immunoassay as an analytical tool in environmental investigations

International Nuclear Information System (INIS)

Van Emon, J.M.

2000-01-01

Immunoassay methods were initially applied in clinical situations where their sensitivity and selectivity were utilized for diagnostic purposes. In the 1970s, pesticide chemists realized the potential benefits of immunoassay methods for compounds difficult to analyze by gas chromatography. This transition of the technology has extended to the analysis of soil, water, food and other matrices of environmental and human exposure significance particularly for compounds difficult to analyze by chromatographic methods. The utility of radioimmunoassays and enzyme immunoassays for environmental investigations was recognized in the 1980s by the U.S. Environmental Protection Agency (U.S. EPA) with the initiation of an immunoassay development programme. The U.S. Department of Agriculture (USDA) and the U.S. Food and Drug Administration (PDA) have investigated immunoassays for the detection of residues in food both from an inspection and a contamination prevention perspective. Environmental immunoassays are providing rapid screening information as well as quantitative information to fulfill rigorous data quality objectives for monitoring programmes

Catch and measure-mass spectrometry-based immunoassays in biomarker research.

Science.gov (United States)

Weiß, Frederik; van den Berg, Bart H J; Planatscher, Hannes; Pynn, Christopher J; Joos, Thomas O; Poetz, Oliver

2014-05-01

Mass spectrometry-based (MS) methods are effective tools for discovering protein biomarker candidates that can differentiate between physiological and pathophysiological states. Promising candidates are validated in studies comprising large patient cohorts. Here, targeted protein analytics are used to increase sample throughput. Methods involving antibodies, such as sandwich immunoassays or Western blots, are commonly applied at this stage. Highly-specific and sensitive mass spectrometry-based immunoassays that have been established in recent years offer a suitable alternative to sandwich immunoassays for quantifying proteins. Mass Spectrometric ImmunoAssays (MSIA) and Stable Isotope Standards and Capture by Anti-Peptide Antibodies (SISCAPA/iMALDI) are two prominent types of MS-based immunoassays in which the capture is done either at the protein or the peptide level. We present an overview of these emerging types of immunoassays and discuss their suitability for the discovery and validation of protein biomarkers. This article is part of a Special Issue entitled: Biomarkers: A Proteomic Challenge. © 2013.

Magnetic properties of nickel nanowires decorated with cobalt nanoparticles fabricated by two step electrochemical deposition technique

Energy Technology Data Exchange (ETDEWEB)

Maaz, K., E-mail: maaz@impcas.ac.cn [Materials Research Center, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou, 730000 (China); Nanomaterials Research Group, Physics Division, PINSTECH, Nilore, 45650, Islamabad (Pakistan); Duan, J.L. [Materials Research Center, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou, 730000 (China); Karim, S. [Nanomaterials Research Group, Physics Division, PINSTECH, Nilore, 45650, Islamabad (Pakistan); Chen, Y.H.; Yao, H.J.; Mo, D.; Sun, Y.M. [Materials Research Center, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou, 730000 (China); Liu, J., E-mail: j.liu@impcas.ac.cn [Materials Research Center, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou, 730000 (China)

2016-10-01

We demonstrate fabrication and magnetic characterization of novel nanostructures composed of Ni nanowires decorated with Co nanoparticles by two step etching and electrochemical deposition in polycarbonate template. Structural analysis confirmed the formation of nickel nanowires with diameter of 62 nm which are surrounded by cobalt nanoparticles of about 15 nm in diameter. By electron microscopy analyses it is evident that the nanoparticles are distributed on the surface of the nanowires. Analysis of magnetization data indicates that ferromagnetic Ni nanowires exhibit an easy axis of magnetization parallel to the wire long-axis while the angular dependence of coercivity indicates that magnetization reversal occurs through the curling process in these nanowires. An exchange bias accompanied by vertical shift in magnetization was observed below ∼20 K, measured under a cooling field of 1 kOe, which is attributed to the spin interactions between the spin-glass like surface layer and ferromagnetic core of the nanowires and nanoparticles. - Highlights: • Co-decorated Ni nanowires were fabricated by two-step electrodeposition technique. • The nanoparticles are distributed on the surface of nanowires. • Magnetization reversal occurs through the curling process in the nanowires. • Temperature dependent coercivity follows thermal activation model.

Designing novel nano-immunoassays: antibody orientation versus sensitivity

Energy Technology Data Exchange (ETDEWEB)

Puertas, S; Moros, M; Fernandez-Pacheco, R; Ibarra, M R; Grazu, V; De la Fuente, J M, E-mail: vgrazu@unizar.e, E-mail: jmfuente@unizar.e [Instituto de Nanociencia de Aragon (INA), Universidad de Zaragoza, Campus RIo Ebro, EdifIcio I-D, Mariano Esquillor, s/n, 50018 Zaragoza (Spain)

2010-12-01

There is a growing interest in the use of magnetic nanoparticles (MNPs) for their application in quantitative and highly sensitive biosensors. Their use as labels of biological recognition events and their detection by means of some magnetic method constitute a very promising strategy for quantitative high-sensitive lateral-flow assays. In this paper, we report the importance of nanoparticle functionalization for the improvement of sensitivity for a lateral-flow immunoassay. More precisely, we have found that immobilization of IgG anti-hCG through its polysaccharide moieties on MNPs allows more successful recognition of the hCG hormone. Although we have used the detection of hCG as a model in this work, the strategy of binding antibodies to MNPs through its sugar chains reported here is applicable to other antibodies. It has huge potential as it will be very useful for the development of quantitative and high-sensitive lateral-flow assays for its use on human and veterinary, medicine, food and beverage manufacturing, pharmaceutical, medical biologics and personal care product production, environmental remediation, etc.

Alkaline phosphatase labeled SERS active sandwich immunoassay for detection of Escherichia coli

Science.gov (United States)

Bozkurt, Akif Goktug; Buyukgoz, Guluzar Gorkem; Soforoglu, Mehmet; Tamer, Ugur; Suludere, Zekiye; Boyaci, Ismail Hakki

2018-04-01

In this study, a sandwich immunoassay method utilizing enzymatic activity of alkaline phosphatase (ALP) on 5-bromo-4-chloro-3-indolyl phosphate (BCIP) for Escherichia coli (E. coli) detection was developed using surface enhanced Raman spectroscopy (SERS). For this purpose, spherical magnetic gold coated core-shell nanoparticles (MNPs-Au) and rod shape gold nanoparticles (Au-NRs) were synthesized and modified for immunomagnetic separation (IMS) of E. coli from the solution. In order to specify the developed method to ALP activity, Au-NRs were labeled with this enzyme. After successful construction of the immunoassay, BCIP substrate was added to produce the SERS-active product; 5-bromo-4-chloro-3-indole (BCI). A good linearity (R2 = 0.992) was established between the specific SERS intensity of BCI at 600 cm- 1 and logarithmic E. coli concentration in the range of 1.7 × 101-1.7 × 106 cfu mL- 1. LOD and LOQ values were also calculated and found to be 10 cfu mL- 1 and 30 cfu mL- 1, respectively.

Bead-based immunoassays

NARCIS (Netherlands)

Wal, van der F.J.; Bergervoet, J.H.W.; Achterberg, R.P.; Haasnoot, W.

2014-01-01

Since the first immunoassay with (radioactive) labeled antibodies in the middle of the 20th century [1], many different formats on various platforms have been developed, using antibodies for capture and/or detection. If antibodies are used to capture compounds, a support, such as the wall of a

Streptavidin-functionalized capillary immune microreactor for highly efficient chemiluminescent immunoassay

Energy Technology Data Exchange (ETDEWEB)

Yang Zhanjun [State Key Laboratory of Analytical Chemistry for Life Science, Department of Chemistry, Nanjing University, Nanjing 210093 (China); College of Chemistry and Engineering, Yangzhou University, 88 South University Avenue, Yangzhou 225002 (China); Zong Chen [State Key Laboratory of Analytical Chemistry for Life Science, Department of Chemistry, Nanjing University, Nanjing 210093 (China); Ju Huangxian, E-mail: hxju@nju.edu.cn [State Key Laboratory of Analytical Chemistry for Life Science, Department of Chemistry, Nanjing University, Nanjing 210093 (China); Yan Feng, E-mail: yanfeng2007@sohu.com [Jiangsu Institute of Cancer Prevention and Cure, Nanjing 210009 (China)

2011-11-07

Highlights: {yields} A novel capillary immune microreactor was proposed for highly efficient flow-through chemiluminescent immunoassay. {yields} The microreactor was prepared by functionalizing capillary inner wall with streptavidin for capture of biotinylated antibody. {yields} The proposed immunoassay method showed wide dynamic range, good reproducibility, stability and practicality. {yields} The microreactor was low-cost and disposable, and possessed several advantages over the conventional immunoreactors. - Abstract: A streptavidin functionalized capillary immune microreactor was designed for highly efficient flow-through chemiluminescent (CL) immunoassay. The functionalized capillary could be used as both a support for highly efficient immobilization of antibody and a flow cell for flow-through immunoassay. The functionalized inner wall and the capture process were characterized using scanning electron microscopy. Compared to conventional packed tube or thin-layer cell immunoreactor, the proposed microreactor showed remarkable properties such as lower cost, simpler fabrication, better practicality and wider dynamic range for fast CL immunoassay with good reproducibility and stability. Using {alpha}-fetoprotein as model analyte, the highly efficient CL flow-through immunoassay system showed a linear range of 3 orders of magnitude from 0.5 to 200 ng mL{sup -1} and a low detection limit of 0.1 ng mL{sup -1}. The capillary immune microreactor could make up the shortcoming of conventional CL immunoreactors and provided a promising alternative for highly efficient flow-injection immunoassay.

Synthesis, characterization, magnetic and electrochemical properties of a new 3D hexa-copper-substituted germanotungstate

International Nuclear Information System (INIS)

Li, Yanzhou; Luo, Jie; Zhang, Yanting; Zhao, Junwei; Chen, Lijuan; Ma, Pengtao; Niu, Jingyang

2013-01-01

An inorganic–organic hybrid hexa-copper-substituted germanotungstate Na 2 [Cu(dap) 2 ] 2 [Cu(dap) 2 ] ([Cu 6 (H 2 O) 2 (dap) 2 ][B-α-GeW 9 O 34 ] 2 )·4H 2 O (1) (dap=1,2-diaminopropane) has been hydrothermally prepared and characterized by elemental analyses, inductively coupled plasma atomic emission spectrometry (ICP–AES) analyses, IR spectra, powder X-ray diffraction (PXRD), thermogravimetric analysis (TGA) and single-crystal X-ray diffraction. 1 displays the six-connected 3D network with the long topological (O′Keefe) vertex symbol is 4·4·6 4 ·4·4·4·4·6 4 ·4·4·4·6 4 ·4·4·4 and the short vertex (Schläfli) symbol of 4 12 6 3 . Magnetic measurements indicate that there are the overall ferromagnetic exchange interactions in the belt-like hexa-Cu II cluster in 1. Furthermore, the electrochemical behavior and electrocatalysis of 1 modified carbon paste electrode (1-CPE) have been studied. The reductions of nitrite, bromate and hydrogen peroxide are principally mediated by the W VI -based wave. - Graphical abstract: A hexa-Cu II sandwiched germanotungstate has been synthesized and structurally characterized. The magnetic, solid-state electrochemical and electrocatalytic properties have been investigated. Display Omitted - Highlights: • Transition-metal substituted polyoxometalates. • Hexa-copper-substituted germanotungstate. • Six-connected 3D network. • Electrocatalytic reduction of nitrite, bromate and hydrogen peroxide

Comparison of capillary electrophoresis-based immunoassay with fluorescence polarization immunoassay for the immunodetermination of methamphetamine using various methamphetamine antibodies.

Science.gov (United States)

Choi, J; Kim, C; Choi, M J

1998-11-01

An accurate and simple immunoassay using capillary electrophoresis (CE) with laser-induced fluorescence (LIF) was performed for the detection of methamphetamine (MA) in urine. The CE-LIF was conducted with an untreated fused-silica column using antiserum and a tracer of fluorescein isothiocyanate (FITC)-labeled MA. This CE-LIF system was compared with fluorescence polarization immunoassay (FPIA) in a TDx analyzer in the photo-check mode using the same FITC-labeled tracer and the same antiserum. Various antibodies, not only those prepared by our own immunogens but also those from commercial sources, were screened and characterized in both assay systems with regard to sensitivity, precision, and cross-reactivity. Both systems satisfied analytical precision and gave similar cross-reactivity patterns. However, the CE-LIF-based immunoassay was approximately one order superior to FPIA in sensitivity, requiring less volume of sample, antiserum, and tracer for the assay. Considering that the FPIA system is well known to be a useful tool for screening antibodies and detecting drugs, the CE-LIF-based immunoassay system, which is seemingly more advantageous than the FPIA system, appears to have great power for the characterization of antibodies and for the detection of MA in urine.

Effect of magnetic field on the microstructure and electrochemical performance of rapidly quenched La{sub 0.1}Nd{sub 0.075}Mg{sub 0.04}Ni{sub 0.65}Co{sub 0.12} alloy

Energy Technology Data Exchange (ETDEWEB)

Chen, Xiangrong [College of Chemistry and Chemical Engineering, Central South University, Changsha 410083 (China); Wang, Haiyan, E-mail: wanghy419@126.com [College of Chemistry and Chemical Engineering, Central South University, Changsha 410083 (China); Hunan Dahua New Energy Co., Ltd., Changsha 410600 (China); Zhu, Shuping; Li, Fangfang [College of Chemistry and Chemical Engineering, Central South University, Changsha 410083 (China); Tang, Yougen, E-mail: ygtang@csu.edu.cn [College of Chemistry and Chemical Engineering, Central South University, Changsha 410083 (China); Liu, Zuming [State Key Laboratory for Powder Metallurgy, Central South University, Changsha 410083 (China)

2014-12-25

Highlights: • La{sub 0.1}Nd{sub 0.075}Mg{sub 0.04}Ni{sub 0.65}Co{sub 0.12} alloy is rapidly quenched in a 0.18 T static magnetic field. • The multiphase structures of as-treated alloys remain unchanged. • Grain refinement is achieved with the aid of magnetic field. • Magnetic field favors the formation of La{sub 2}Ni{sub 7} phase. • The as-prepared alloy exhibits improved electrochemical performance. - Abstract: Rare earth–Mg–Ni-based (RE–Mg–Ni-based) La{sub 0.1}Nd{sub 0.075}Mg{sub 0.04}Ni{sub 0.65}Co{sub 0.12} hydrogen storage alloys were rapidly quenched with and without exerting a 0.18 T static magnetic field and investigated by X-ray diffraction (XRD), scanning electron microscope (SEM), energy dispersive spectroscopy (EDS) studies and various electrochemical measurements. The results show that all samples hold a two-phase structure consisting of La{sub 2}Ni{sub 7} phase and LaNi{sub 5} phase, suggesting that the structure remains unchanged after treatment. Grain refinement, homogeneous composition and increase in La{sub 2}Ni{sub 7} phase abundance are achieved when magnetic field is applied. In comparison to quenched alloys, higher discharge capacities are obtained for the alloys prepared with the aid of magnetic field mainly due to the larger La{sub 2}Ni{sub 7} phase abundance. Cycling stability is improved with increasing quenching rate probably owing to better anti-pulverization ability resulted from refined grain size. Ameliorated electrochemical kinetics of the magnetic field assisted rapidly quenched alloys has been confirmed by potential-step measurements and electrochemical impedance spectroscopy (EIS) tests in accordance with the enhanced electrochemical properties.

Effects of magnetic processing on electrochemical and photoelectrochemical properties of electrodes modified with nanoclusters of a phenothiazine-C{sub 60} linked compound

Energy Technology Data Exchange (ETDEWEB)

Yonemura, H; Yamada, S [Department of Applied Chemistry, Faculty of Engineering, Kyushu University, Fukuoka 819-0395 (Japan); Wakita, Y; Moribe, S [Department of Materials Physics and Chemistry, Graduate School of Engineering, Kyushu University, Fukuoka 819-0395 (Japan); Fujiwara, Y; Tanimoto, Y [Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Higashi-Hiroshima, Hiroshima 739-8526 (Japan)], E-mail: yonemura@mail.cstm.kyushu-u.ac.jp

2009-03-01

Effects of magnetic processing on morphological, electrochemical, and photoelectrochemical properties of electrodes modified with nanoclusters of a phenothiazine-C{sub 60} linked compound with four methylene group (Ph(4)C{sub 60}) were examined in the absence and presence of magnetic processing with three different magnetic environments due to strong magnetic field. The AFM measurements indicated that the morphologies of nanostructures of Ph(4)C{sub 60} varied with magnetic enviroments as comparison with that in the absence of magnetic processing. At top position (5.6 T; - 940 T{sup 2}/m) with hypogravity, large spherical nanoclusters (60{approx}70 nm diameter) were observed as comparion with those (ca. 20 nm diameter) in the absence of magnetic processing. At middle positon (15 T; 0 T{sup 2}/m) with normal gravity, the fiber-like nanostructure was observed. At bottom position (9.8 T; + 1070T{sup 2}/m) with hypergravity, the rod-like nanostrucure was observed. The interesting results might be ascribed to the different solvent properties due to the different rates of evaporation of two solvents in the toluene-acetonitrile mixed solvent during drying process under various magnetic environments. First reduction peaks due to C{sub 60} moiety of Ph(4)C{sub 60} nanostrucures in the presence of magnetic processing at three different positions were negative-shifted as comparison with that in the absence of magnetic processing. Potential dependencies of the photocurrents of the electrodes modified with Ph(4)C{sub 60} nanostrucures in the presence of magnetic processing at three positions were also different from that in the absence of magnetic processing. The magnetic field effects in AFM, and electrochemical and photoelectrochemical measurements are most likely ascribed to the difference of the reduction potentials due to C{sub 60} clusters between the absence and presence of magnetic processing due to the morphological change of Ph(4)C{sub 60} nanostrucures.

Electrochemical genosensing of Salmonella, Listeria and Escherichia coli on silica magnetic particles.

Science.gov (United States)

Liébana, Susana; Brandão, Delfina; Cortés, Pilar; Campoy, Susana; Alegret, Salvador; Pividori, María Isabel

2016-01-21

A magneto-genosensing approach for the detection of the three most common pathogenic bacteria in food safety, such as Salmonella, Listeria and Escherichia coli is presented. The methodology is based on the detection of the tagged amplified DNA obtained by single-tagging PCR with a set of specific primers for each pathogen, followed by electrochemical magneto-genosensing on silica magnetic particles. A set of primers were selected for the amplification of the invA (278 bp), prfA (217 bp) and eaeA (151 bp) being one of the primers for each set tagged with fluorescein, biotin and digoxigenin coding for Salmonella enterica, Listeria monocytogenes and E. coli, respectively. The single-tagged amplicons were then immobilized on silica MPs based on the nucleic acid-binding properties of silica particles in the presence of the chaotropic agent as guanidinium thiocyanate. The assessment of the silica MPs as a platform for electrochemical magneto-genosensing is described, including the main parameters to selectively attach longer dsDNA fragments instead of shorter ssDNA primers based on their negative charge density of the sugar-phosphate backbone. This approach resulted to be a promising detection tool with sensing features of rapidity and sensitivity very suitable to be implemented on DNA biosensors and microfluidic platforms. Copyright © 2015 Elsevier B.V. All rights reserved.

Recent advancements in the immunoassay domain

International Nuclear Information System (INIS)

Pradelles, Ph.

1997-01-01

The two types of immunoassay techniques, the competition analysis and the immuno-metric analysis (sandwich type), are described; the tracers used with theses methods have high specific radioactivity levels in order to be traced at extremely low content. Non radioactive tracers have been also developed, such as enzymatic, fluorescent, luminescent tracers, which are simpler and may be used at home. The Cea has recently developed some innovative immunoassay formats, such as acetylcholinesterase as a new enzymatic tracer, and immuno-metric dosage for very small molecules such as haptenes

An ultrasensitive chemiluminescence immunoassay for fumonisin B1 detection in cereals based on gold-coated magnetic nanoparticles.

Science.gov (United States)

Jie, Mingsha; Yu, Songcheng; Yu, Fei; Liu, Lie; He, Leiliang; Li, Yanqiang; Zhang, Hongquan; Qu, Lingbo; Harrington, Peter de B; Wu, Yongjun

2018-07-01

In the present study, a novel highly sensitive magnetic enzyme chemiluminescence immunoassay (MECLIA) was developed to detect fumonisin B 1 (FB 1 ) in cereal samples. The gold-coated magnetic nanoparticles (Fe 3 O 4 @Au, GoldMag) were used as solid phase carrier to develop a competitive CLIA for detecting FB 1 , in which FB 1 in samples would compete with FB 1 -ovalbumin coated on the surface of Fe 3 O 4 @Au nanoparticles for binding with FB 1 antibodies. Successively, horseradish peroxidase labeled goat anti-rabbit IgG (HRP-IgG) was conjugated with FB 1 antibodies on the microplate. In substrate solution containing luminol and H 2 O 2 , HRP-IgG catalyzed luminol oxidation by H 2 O 2 , generating a high chemiluminescence signal. The FB 1 immune GoldMag particles were characterized by Fourier transform infrared spectroscopy, scanning electron microscope and zeta potential analysis, etc. RESULTS: The concentrations and the reaction times of these immunoreagents were optimized to improve the performances of this method. The established method could detect as low as 0.027 ng mL -1 FB 1 from 0.05 ng mL -1 to 25 ng mL -1 , demonstrating little cross-reaction (less than 2.4%) with other structurally related compounds. The average intrassay relative SD (RSD) (n = 6) was 3.4% and the average interassay RSD (n = 6) was 5.4%. This method was successfully applied for the determination of FB 1 in corn and wheat and gave recoveries of between 98-110% and 91-105%, respectively. The results of the present study suggest that the MECLIA approach has potential application for high-throughput fumonisin screening in cereals. © 2018 Society of Chemical Industry. © 2018 Society of Chemical Industry.

Synthesis of magnetite nanoparticles using electrochemical oxidation

Directory of Open Access Journals (Sweden)

Ye. Ya. Levitin

2014-08-01

Full Text Available The monodisperse magnetite nanoparticles are promising for use in the biomedical industry for targeted drug delivery, cell separation and biochemical products, Magnetic Resonance Imaging, immunological studies, etc. Classic method for the synthesis of magnetite is the chemical condensation Elmore’s, it is simple and cheap, but it is complicated by the formation of side compounds which impair the magnetic properties of the final product. Biological and medical purposes require high purity magnetite nanoparticles. Electrochemical methods of producing nanoparticles of magnetite acquire significant spread. The kinetics of electrochemical processes are a function of a larger number of parameters than the kinetics of conventional chemical reaction, thus electrochemical reactions can be thinner and more completely adjusted to give a predetermined size nanoparticles. In the kinetics of the electrochemical oxidation and reduction the important role is played by the nature of the electrode. In many industrial processes, it is advisable to use lead dioxide anodes with titanium current lead. Purpose of the work To determine the optimum conditions of electrochemical oxidation of Fe2+ Fe3+to produce magnetite with high purity and improved magnetic characteristics. Materials and methods Electrochemical studies were carried out in a glass cell ЯСЭ-2 using a potentiostat ПИ-50-1.1 and a recording device ПДА1. Reference electrode - silver chloride ЭВЛ1М 3.1, potentials listed on the hydrogen scale. The test solution contained 80 g/ l FeSO4×7H2O and H2SO4(to pH 1. The pH of the solution was measured with a pH–meter « рН–150». Concentration ratio of Fe3+/Fe2+in the solution was measured by permanganometric method. Magnetite particle sizes were measured by an electron microscope computer ЭВМ-100Л, an increasing is 2×105. Saturation magnetization was evaluated by the magnetization curve, for the measured sample in the field with strength

Review of the biochemical basis of enzyme immunoassays

International Nuclear Information System (INIS)

Klingler, W.

1982-01-01

The ever increasing number of radioimmunological determination poses problems allied with the handling of radioactive substances. In recent years various non-radioactive methods have been developed, among which the enzyme immunoassay is already in routine use. Homogeneous and heterogeneous enzyme immunoassays are described. Criteria for enzymes, substrates and enzyme-substrate reactions are listed. (orig.) [de

Ultrasensitive molecularly imprinted electrochemical sensor based on magnetism graphene oxide/β-cyclodextrin/Au nanoparticles composites for chrysoidine analysis

International Nuclear Information System (INIS)

Wang, Xiaojiao; Li, Xiangjun; Luo, Chuannan; Sun, Min; Li, Leilei; Duan, Huimin

2014-01-01

Highlights: • Synthesis and application of MGO/β-CD@AuNPs as a sensor for chrysoidine analysis. • The synthesized polymer had a laminar structure with high surface. • The propose sensor showed high selectivity and good sensitivity. - Abstract: A imprinted electrochemical sensor based on glassy carbon electrode (GCE) for ultrasensitive detection of chrysoidine was fabricated. A GCE was modified by magnetic graphene oxide/β-cyclodextrin/gold nanoparticles composites (MGO/β-CD@AuNPs). The sensing surface area and electronic transmission rate were increased, which was benefited from the distribution property of MGO/β-CD@AuNPs. The MGO/β-CD@AuNPs composite improved electrochemical response and sensitivity of the sensor. The molecularly imprinted electrochemical sensor was prepared by electropolymerization on modified electrode. Chrysoidine and pyrrole were used as template molecule and functional monomer, respectively. Under the optimization experimental conditions, the electrochemical sensor exhibited excellent analytical performance: the detection of chrysoidine ranged from 5.0 × 10 −8 mol/L to 5.0 × 10 −6 mol/L with the detection limit of 1.7 × 10 −8 mol/L. The sensor was applied to determine chrysoidine in spiked water samples and showed high selectivity, good sensitivity and acceptable reproducibility. The proposed method provides a promising platform for trace amount detection of other food additives

Process for electrochemically gasifying coal using electromagnetism

Science.gov (United States)

Botts, Thomas E.; Powell, James R.

1987-01-01

A process for electrochemically gasifying coal by establishing a flowing stream of coal particulate slurry, electrolyte and electrode members through a transverse magnetic field that has sufficient strength to polarize the electrode members, thereby causing them to operate in combination with the electrolyte to electrochemically reduce the coal particulate in the slurry. Such electrochemical reduction of the coal produces hydrogen and carbon dioxide at opposite ends of the polarized electrode members. Gas collection means are operated in conjunction with the process to collect the evolved gases as they rise from the slurry and electrolyte solution.

Chemiluminescence immunoassay for chloramphenicol

International Nuclear Information System (INIS)

Lin Si; Xu Wenge; Liu Yibing

2007-06-01

A simple, solid-phase chemiluminescence immunoassay (CLIA) for the measurement of Chloramphenicol(CAP) in foodstuffs is described. A rabbit anti-CAP IgG is passively adsorbed onto the walls of polypropylene plates. The labeled conjugant is horseradish peroxidase(HRP) conjugate of CAP. Luminol solution is used as the substrate of HRP. The light yield is inversely proportional to the concentration of CAP. The method has a similar sensitivity (0.05 ng/mL), specificity, precision, and accuracy to a conventional enzyme immunoassay (EIA). The intra-assay and inter-assay CVs of ten samples were <8 and <20%, respectively, and the analytical recovery of the method was 87% 100%. The experimental correlation coefficient of dilution was found to be 0.999 using milk supernatant as buffer. The assay range for the method was 0.1-10 ng/mL, and it displayed good linearity. (authors)

The immunoassay handbook

National Research Council Canada - National Science Library

Wild, David (David G.)

2001-01-01

... importantly, enabling them to keep current on the basic theory behind immunoassay. Since the publication of the previous edition in 1994, the field has continued to evolve rapidly, and the need for a fully updated version of this book is now paramount. The second edition has been comprehensively updated and new chapters have been added to each section" [publisher's web site].

Evaluation of Six Different Immunoassays for Serum Thyrotropin

International Nuclear Information System (INIS)

Ma Donghong; Lu Hankui; Gao Yunchao; Ge Wenli; Xiong Jiang; Liu Qiaoping; Gu Qing

2010-01-01

To analyzes the discrepancy and association among six different thyrotropin (TSH) immunoassay methods and to study their impact on the clinical diagnoses of thyroid diseases, the 150 serum samples from three groups consisting of hyperthyroidism, hypothyroidism and healthy subjects, 50 samples in each group were included in this study. The serum TSH levels were measured simultaneously by radioimmunoassay (RIA), immunoradiometric assay (IRMA), three-type chemilumiminescence immunoassay (CLIA) and electrochemiluminescence immunoassay (ECLIA). The results showed that individual serum TSH level varied significantly from one assay to another. There was no correlation between TSH RIA and other five assays in groups of hyperthyroidism and healthy subjects(P>0.05). The correlations between TSH IRMA and four automatic assays in hyperthyroidism group were relatively low (r= 0.38∼0.41). However, among the four automatic assays, TSH levels were well correlated (r= 0.92∼0.99). For clinical diagnoses, TSH RIA alone was not useful in the differentiation of hyperthyroidism and normal subjects, and TSH IRMA was misleading in some hyperthyroidism. There were no significant differences for four TSH automatic immunoassays in differential diagnoses of thyroid diseases. (authors)

Multiplex detection of plant pathogens using a microsphere immunoassay technology.

Directory of Open Access Journals (Sweden)

Ratthaphol Charlermroj

Full Text Available Plant pathogens are a serious problem for seed export, plant disease control and plant quarantine. Rapid and accurate screening tests are urgently required to protect and prevent plant diseases spreading worldwide. A novel multiplex detection method was developed based on microsphere immunoassays to simultaneously detect four important plant pathogens: a fruit blotch bacterium Acidovorax avenae subsp. citrulli (Aac, chilli vein-banding mottle virus (CVbMV, potyvirus, watermelon silver mottle virus (WSMoV, tospovirus serogroup IV and melon yellow spot virus (MYSV, tospovirus. An antibody for each plant pathogen was linked on a fluorescence-coded magnetic microsphere set which was used to capture corresponding pathogen. The presence of pathogens was detected by R-phycoerythrin (RPE-labeled antibodies specific to the pathogens. The assay conditions were optimized by identifying appropriate antibody pairs, blocking buffer, concentration of RPE-labeled antibodies and assay time. Once conditions were optimized, the assay was able to detect all four plant pathogens precisely and accurately with substantially higher sensitivity than enzyme-linked immunosorbent assay (ELISA when spiked in buffer and in healthy watermelon leaf extract. The assay time of the microsphere immunoassay (1 hour was much shorter than that of ELISA (4 hours. This system was also shown to be capable of detecting the pathogens in naturally infected plant samples and is a major advancement in plant pathogen detection.

Multiplex detection of plant pathogens using a microsphere immunoassay technology.

Science.gov (United States)

Charlermroj, Ratthaphol; Himananto, Orawan; Seepiban, Channarong; Kumpoosiri, Mallika; Warin, Nuchnard; Oplatowska, Michalina; Gajanandana, Oraprapai; Grant, Irene R; Karoonuthaisiri, Nitsara; Elliott, Christopher T

2013-01-01

Plant pathogens are a serious problem for seed export, plant disease control and plant quarantine. Rapid and accurate screening tests are urgently required to protect and prevent plant diseases spreading worldwide. A novel multiplex detection method was developed based on microsphere immunoassays to simultaneously detect four important plant pathogens: a fruit blotch bacterium Acidovorax avenae subsp. citrulli (Aac), chilli vein-banding mottle virus (CVbMV, potyvirus), watermelon silver mottle virus (WSMoV, tospovirus serogroup IV) and melon yellow spot virus (MYSV, tospovirus). An antibody for each plant pathogen was linked on a fluorescence-coded magnetic microsphere set which was used to capture corresponding pathogen. The presence of pathogens was detected by R-phycoerythrin (RPE)-labeled antibodies specific to the pathogens. The assay conditions were optimized by identifying appropriate antibody pairs, blocking buffer, concentration of RPE-labeled antibodies and assay time. Once conditions were optimized, the assay was able to detect all four plant pathogens precisely and accurately with substantially higher sensitivity than enzyme-linked immunosorbent assay (ELISA) when spiked in buffer and in healthy watermelon leaf extract. The assay time of the microsphere immunoassay (1 hour) was much shorter than that of ELISA (4 hours). This system was also shown to be capable of detecting the pathogens in naturally infected plant samples and is a major advancement in plant pathogen detection.

Nanobody medicated immunoassay for ultrasensitive detection of cancer biomarker alpha-fetoprotein.

Science.gov (United States)

Chen, Jing; He, Qing-hua; Xu, Yang; Fu, Jin-heng; Li, Yan-ping; Tu, Zhui; Wang, Dan; Shu, Mei; Qiu, Yu-lou; Yang, Hong-wei; Liu, Yuan-yuan

2016-01-15

Immunoassay for cancer biomarkers plays an important role in cancer prevention and early diagnosis. To the development of immunoassay, the quality and stability of applied antibody is one of the key points to obtain reliability and high sensitivity for immunoassay. The main purpose of this study was to develop a novel immunoassay for ultrasensitive detection of cancer biomarker alpha-fetoprotein (AFP) based on nanobody against AFP. Two nanobodies which bind to AFP were selected from a phage display nanobody library by biopanning strategy. The prepared nanobodies are clonable, thermally stable and applied in both sandwich enzyme linked immunoassay (ELISA) and immuno-PCR assay for ultrasensitive detection of AFP. The limit detection of sandwich ELISA setup with optimized nanobodies was 0.48ng mL(-1), and the half of saturation concentration (SC50) value was 6.68±0.56ng mL(-1). These nanobodies were also used to develop an immuno-PCR assay for ultrasensitive detection of AFP, its limit detection values was 0.005ng mL(-1), and the linear range was 0.01-10,000ng mL(-1). These established immunoassays based on nanobodies were highly specific to AFP and with negligible cross reactivity with other tested caner biomarkers. Furthermore, this novel concept of nanobodies mediated immunoassay may provide potential applications in a general method for the ultrasensitive detection of various cancer biomarkers. Copyright © 2015 Elsevier B.V. All rights reserved.

Magnetic particles in medical research - a review

International Nuclear Information System (INIS)

Sajid, K.M.

2001-01-01

Magnetic (or magnetizable) particles have assumed increasing importance in medical and biological research since 1966 when the effect of a magnetic field on the movement of suspended particles was initially studied. In fields like haematology, cell biology, microbiology, biochemistry and immunoassays, they currently provide the basis for separation techniques, which previously relied on gravitational forces. The body cells (e.g., blood cells) can be made magnetic by incubating them in a medium containing several Fe/sub 3/O/sub 4/ particles, which are adsorbed to the membrane surfaces. Some bacteria (also called magnetostatic bacteria) respond to externally applied magnetic lines of force due to their intracellular magnetic particles. These properties are useful in the isolation of these cells/bacteria. In biochemistry magnetic particles are used to immobilize enzymes without any loss of enzyme activity. The immobilized enzymes can facilitate the separation of end products without extensive instrumentation. In immunoassays the antibodies are covalently linked to polymer coated iron oxide particles. An electromagnet is used to sediment these particles after reaction. This excludes the use of centrifuge to separate antigen-antibody complexes. In pharmacy and pharmacology the magnetic particles are important in drug transport. In techniques like ferrography, nuclear magnetic resonance imaging (NMRI), spectroscopic studies and magnetic resonance imaging (MRI) the magnetic particles serve as contrast agents and give clinically important spatial resolution. Magnetic particles also find extensive applications in cancer therapy, genetic engineering, pneumology, nuclear medicine, radiology and many other fields. This article reviews these applications. (author)

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

Tunable electronic and magnetism of SrTiO{sub 3}/BiFeO{sub 3} (001) superlattice: For electrochemical applications

Energy Technology Data Exchange (ETDEWEB)

Xu, Qiang; Sopiha, Kostiantyn; Sobhan, Mushtaq; Anariba, Franklin; Wu, Ping, E-mail: wuping@sutd.edu.sg [Entropic Interface Group (EIG), Engineering Product Development, Singapore University of Technology and Design (SUTD), 8 Somapah Road, Singapore 487372 (Singapore); Ong, Khuong Phuong; Zheng, Jian Wei [Institute of High Performance Computing (IHPC), Agency for Science, Technology and Research - A*Star, 1 Fusionopolis Way, #16-16 Connexis, Singapore 138632 (Singapore)

2016-01-04

Practical strategy in tuning the conductivity and magnetism of SrTiO{sub 3}/BiFeO{sub 3} (STO/BFO) (001) superlattice is investigated using the first-principles method based on density functional theory. Our calculated results show that both the conductivity and magnetism of this superlattice can be tuned via a control of its interface terminations. The STO layers maintain semiconducting, while the BFO layers demonstrate metallic character. Therefore, the conductivity of STO/BFO is controlled by the BFO layers. Furthermore, a magnetic STO/BFO (001) superlattice can be found in n-type TiO{sub 2}/BiO interface but with heavy electron carriers. The thickness of BFO does not change the electronic structure and character of STO/BFO (001) superlattice. This study provides a fundamental understanding of the chemically turned conductivity and magnetism of BFO thin films, which may further advance electrochemical applications like magnetic-field aided chemical gas sensing, solar cells, and photo-catalytic chemical reactions.

Effective gasoline site assessment using the D TECH trademark BTEX immunoassay

International Nuclear Information System (INIS)

Hudak, R.T.; Melby, J.M.; Stave, J.W.

1994-01-01

The application of immunoassay to environmental testing can greatly aid in assessing sites for various contaminants including PCB, TNT, RDX, PAH and BTEX. Immunoassay offers several benefits to site assessment: it is accurate, reproducible and relatively inexpensive compared to instrumental analysis. In addition, because of its ease-of-use, this technique is ideal for the field and requires minimal user training. To demonstrate not only the effectiveness of the BTEX immunoassay, but also the reliability of the field results, a gasoline contaminated site was assessed comparing the BTEX immunoassay to gas chromatography. All sampling and site related activities were executed in accordance to the USEPA SW-846 guidelines. Three (3) analyses were performed on each sample. One immunoassay analysis was performed in the field by an individual who received two (2) hours of training prior to the start of the study. A technician familiar with the immunoassay ran the second analysis in a laboratory. Finally, an independent GC laboratory certified for BTEX method 8020 and 602 performed the GC analyses. One hundred one (101) samples were analyzed: thirty-nine (39) samples were water, the other sixty-two (62) were soils ranging from clay to silt. The results and costs of the methods are compared

Synthesis, characterization, magnetic and electrochemical properties of a new 3D hexa-copper-substituted germanotungstate

Energy Technology Data Exchange (ETDEWEB)

Li, Yanzhou; Luo, Jie; Zhang, Yanting [Institute of Molecular and Crystal Engineering, Henan Key Lab of Polyoxometalate Chemistry, College of Chemistry and Chemical Engineering, Henan University, Kaifeng, Henan 475004 (China); Zhao, Junwei, E-mail: zhaojunwei@henu.edu.cn [Institute of Molecular and Crystal Engineering, Henan Key Lab of Polyoxometalate Chemistry, College of Chemistry and Chemical Engineering, Henan University, Kaifeng, Henan 475004 (China); Basic Experiment Teaching Center, Henan University, Kaifeng, Henan 475004 (China); Chen, Lijuan, E-mail: ljchen@henu.edu.cn [Institute of Molecular and Crystal Engineering, Henan Key Lab of Polyoxometalate Chemistry, College of Chemistry and Chemical Engineering, Henan University, Kaifeng, Henan 475004 (China); State Key Laboratory of Structural Chemistry, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, Fujian 350002 (China); Ma, Pengtao; Niu, Jingyang [Institute of Molecular and Crystal Engineering, Henan Key Lab of Polyoxometalate Chemistry, College of Chemistry and Chemical Engineering, Henan University, Kaifeng, Henan 475004 (China)

2013-09-15

An inorganic–organic hybrid hexa-copper-substituted germanotungstate Na{sub 2}[Cu(dap){sub 2}]{sub 2}[Cu(dap){sub 2}] ([Cu{sub 6}(H{sub 2}O){sub 2}(dap){sub 2}][B-α-GeW{sub 9}O{sub 34}]{sub 2})·4H{sub 2}O (1) (dap=1,2-diaminopropane) has been hydrothermally prepared and characterized by elemental analyses, inductively coupled plasma atomic emission spectrometry (ICP–AES) analyses, IR spectra, powder X-ray diffraction (PXRD), thermogravimetric analysis (TGA) and single-crystal X-ray diffraction. 1 displays the six-connected 3D network with the long topological (O′Keefe) vertex symbol is 4·4·6{sub 4}·4·4·4·4·6{sub 4}·4·4·4·6{sub 4}·4·4·4 and the short vertex (Schläfli) symbol of 4{sup 12}6{sup 3}. Magnetic measurements indicate that there are the overall ferromagnetic exchange interactions in the belt-like hexa-Cu{sup II} cluster in 1. Furthermore, the electrochemical behavior and electrocatalysis of 1 modified carbon paste electrode (1-CPE) have been studied. The reductions of nitrite, bromate and hydrogen peroxide are principally mediated by the W{sup VI}-based wave. - Graphical abstract: A hexa-Cu{sup II} sandwiched germanotungstate has been synthesized and structurally characterized. The magnetic, solid-state electrochemical and electrocatalytic properties have been investigated. Display Omitted - Highlights: • Transition-metal substituted polyoxometalates. • Hexa-copper-substituted germanotungstate. • Six-connected 3D network. • Electrocatalytic reduction of nitrite, bromate and hydrogen peroxide.

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

Nanosilver-penetrated polyion graphene complex membrane for mediator-free amperometric immunoassay of alpha-fetoprotein using nanosilver-coated silica nanoparticles

International Nuclear Information System (INIS)

Tang Juan; Tang Dianping; Su Biling; Li Qunfang; Qiu Bin; Chen Guonan

2011-01-01

Research highlights: → We fabricate a polyion graphene complex membrane-based immunosensing platform for sensitive electrochemical immunoassay of alpha-fetoprotein. → Nanosilver-coated silica nanocomposites as bionanolabels. → Graphene nanosheets, single-stranded DNA and silver nanoparticles as matrices. → Direct electron transfer without electron mediator. → 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 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 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 2 (HRP-anti-AFP-Ag-SiO 2 ) as secondary antibodies. Compared with pure silver nanoparticles, Ag-SiO 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 3σ) in pH 6.0 PBS-H 2 O 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 electrochemiluminescent analyzer.

Three-dimensional nanoflower-like MnO2 functionalized graphene as catalytically promoted nanolabels for ultrasensitive electrochemiluminescence immunoassay

International Nuclear Information System (INIS)

Su, Min; Zhang, Yan; Song, Xianrang; Ge, Shenguang; Yan, Mei; Yu, Jinghua; Huang, Jiadong

2013-01-01

A novel electrochemiluminescence (ECL) immunosensor based on the peroxydisulfate solution for detection of prostate-specific antigen (PSA) has been developed. In this work, gold nanoparticles (AuNPs) were electrodeposited on the glassy carbon electrode surface, which could increase the surface area to capture a large amount of primary antibodies as well as to improve the electronic transmission rate. The as-prepared bionanolabels, three-dimensional (3D) nanoflower-like MnO 2 functionalized graphene (GN/MnO 2 ), with novel hybrid architecture were initially fabricated via electrostatic interaction. Then, to provide an effective matrix for antibody immobilization with good stability and bioactivity, the novel materials were modified by AuNPs with the aid of poly (diallyldimethylammonium chloride) via a simple sonication-induced assembly method. In this immunosensor, MnO 2 with the characteristics of well electrochemical behavior, low cost, and environmental-friendly, is used as a substitute for horseradish peroxidase to catalyze H 2 O 2 to produce O 2 , and its 3D nanoflower-like structure is beneficial for the unparallel surface-to-volume ratio. Furthermore, a simple, low-cost and portable home-made potential transformer was utilized in this system to take the place of conventional electrochemical workstation. With a sandwich-type immunoassay format, the amount of functionalized GN/MnO 2 labeled antibodies increased with the increment of antigens in the samples, which implied that the ECL signals enhanced with the increase of in situ generated O 2 due to the enhancive catalysis of MnO 2 to the H 2 O 2 . The immunosensor displayed excellent analytical performance for the detection of PSA in the range of 0.005–10 ng mL −1 with a detection limit of 2.5 pg mL −1 at 3σ. Moreover, the proposed method showed good precision, acceptable stability and reproducibility, and could be used for the detection of PSA in real samples. The proposed method provides a new

Fluorescence immunoassay for detecting periodontal bacterial pathogens in plaque.

OpenAIRE

Wolff, L F; Anderson, L; Sandberg, G P; Aeppli, D M; Shelburne, C E

1991-01-01

A particle concentration fluorescence immunoassay has been modified into a bacterial concentration fluorescence immunoassay (BCFIA) to rapidly detect periodontopathic bacteria in human plaque samples. The BCFIA utilizes fluorescently tagged monoclonal antibodies (MAbs) directed against the lipopolysaccharide of selected gram-negative plaque bacteria. Microorganisms closely associated with periodontal disease that can be identified in plaque with the BCFIA include Porphyromonas gingivalis, Bac...

Fabrication and characterization of tosyl-activated magnetic and nonmagnetic monodisperse microspheres for use in microfluic-based ferritin immunoassay

Czech Academy of Sciences Publication Activity Database

Reymond, F.; Vollet, Ch.; Plichta, Zdeněk; Horák, Daniel

2013-01-01

Roč. 29, č. 2 (2013), s. 532-542 ISSN 8756-7938 R&D Projects: GA MŠk 7E12053; GA ČR GAP503/10/0664 EU Projects: European Commission(XE) 246513 - NADINE Institutional support: RVO:61389013 Keywords : biosensors * electrochemistry * immunoassays Subject RIV: CD - Macromolecular Chemistry Impact factor: 1.883, year: 2013

A half-ring GMR sensor for detection of magnetic beads immobilized on a circular micro-trap

KAUST Repository

Gooneratne, Chinthaka Pasan

2011-11-01

Utilizing magnetic principles in biological immunoassays is an attractive option given its ability to remotely and non-invasively manipulate and detect cells tagged with micro/nano size superparamagnetic type beads and due to the fact that even the most complex biological immunoassays will have very little magnetic effect. The presence of magnetic beads can be detected by a magnetic sensor which quantifies the amount of target cells present in the immunoassay. In order to increase the detection rate a circular conducting micro-trap is employed to attract, trap and transport the magnetic beads to the sensing area. In this research we propose a half-ring spin valve type giant magnetoresistance (GMR) sensor for the measurement of stray fields produced by 2 μm magnetic beads which are around the circular micro-trap. A couple of half-ring GMR sensors can be used to cover the entire circular border width, in order to detect the majority of the immobilized magnetic beads. Analytical and numerical analysis leading towards the fabrication of the half-ring GMR sensor are presented. DC characterization of the fabricated sensor showed a magnetoresistance of 5.9 %. Experimental results showed that the half-ring GMR sensor detected the presence of 2 μm magnetic beads. Hence, half-ring GMR sensors integrated with a circular micro-trap have great potential to be used as an effective disease diagnostic device. © 2011 IEEE.

A half-ring GMR sensor for detection of magnetic beads immobilized on a circular micro-trap

KAUST Repository

Gooneratne, Chinthaka Pasan; Liang, Cai; Useinov, Arthur; Kosel, Jü rgen; Giouroudi, Ioanna

2011-01-01

Utilizing magnetic principles in biological immunoassays is an attractive option given its ability to remotely and non-invasively manipulate and detect cells tagged with micro/nano size superparamagnetic type beads and due to the fact that even the most complex biological immunoassays will have very little magnetic effect. The presence of magnetic beads can be detected by a magnetic sensor which quantifies the amount of target cells present in the immunoassay. In order to increase the detection rate a circular conducting micro-trap is employed to attract, trap and transport the magnetic beads to the sensing area. In this research we propose a half-ring spin valve type giant magnetoresistance (GMR) sensor for the measurement of stray fields produced by 2 μm magnetic beads which are around the circular micro-trap. A couple of half-ring GMR sensors can be used to cover the entire circular border width, in order to detect the majority of the immobilized magnetic beads. Analytical and numerical analysis leading towards the fabrication of the half-ring GMR sensor are presented. DC characterization of the fabricated sensor showed a magnetoresistance of 5.9 %. Experimental results showed that the half-ring GMR sensor detected the presence of 2 μm magnetic beads. Hence, half-ring GMR sensors integrated with a circular micro-trap have great potential to be used as an effective disease diagnostic device. © 2011 IEEE.

Detection of narcotics with an immunoassay film badge

International Nuclear Information System (INIS)

Lukens, H.R.

1993-01-01

Efficient personnel performance, a major requirement for a safe nuclear industry, is jeopardized where personnel use narcotics. However, detection of narcotics at nuclear plants is a challenge. The unique specificity and sensitivity of an immunoassay has been implemented in the form of a small, dry immunoassay film badge (IFB) for the detection of vapors emitted by narcotics. The device is suitable as an area monitor, and its characteristics are suitable for use as a breath monitor for the detection of drug use

Development of national immunoassay reagent programmes

International Nuclear Information System (INIS)

Sufi, S.B.; Micallef, J.V.; Ahsan, R.; Goncharov, N.P.

1992-01-01

Despite the existence of networks of fully equipped laboratories with well-trained staff, the availability of immunodiagnostic services in developing countries is often limited by the high cost of imported kits. There are a number of ways of tackling this problem, ranging from bulk purchase of kits or reagents to local development and production of assay systems. Argentina/Chile, China, Cuba/Mexico, and Thailand are amongst the countries which have established local immunoassay reagent programmes to manufacture low cost, high quality immunoassay reagents. Kits from these projects are now beginning to become available, and it is hoped that they will promote national diagnostic services and research, as well as stimulating the development of reagent programmes for other analytes. (author). 4 refs, 1 tab

Control of (pre-analytical aspects in immunoassay measurements of metabolic hormones in rodents

Directory of Open Access Journals (Sweden)

Maximilian Bielohuby

2018-04-01

Full Text Available The measurement of circulating hormones by immunoassay remains a cornerstone in preclinical endocrine research. For scientists conducting and interpreting immunoassay measurements of rodent samples, the paramount aim usually is to obtain reliable and meaningful measurement data in order to draw conclusions on biological processes. However, the biological variability between samples is not the only variable affecting the readout of an immunoassay measurement and a considerable amount of unwanted or unintended variability can be quickly introduced during the pre-analytical and analytical phase. This review aims to increase the awareness for the factors ‘pre-analytical’ and ‘analytical’ variability particularly in the context of immunoassay measurement of circulating metabolic hormones in rodent samples. In addition, guidance is provided how to gain control over these variables and how to avoid common pitfalls associated with sample collection, processing, storage and measurement. Furthermore, recommendations are given on how to perform a basic validation of novel single and multiplex immunoassays for the measurement of metabolic hormones in rodents. Finally, practical examples from immunoassay measurements of plasma insulin in mice address the factors ‘sampling site and inhalation anesthesia’ as frequent sources of introducing an unwanted variability during the pre-analytical phase. The knowledge about the influence of both types of variability on the immunoassay measurement of circulating hormones as well as strategies to control these variables are crucial, on the one hand, for planning and realization of metabolic rodent studies and, on the other hand, for the generation and interpretation of meaningful immunoassay data from rodent samples.

Structural and magnetic properties of nickel nanowires grown in porous anodic aluminium oxide template by electrochemical deposition technique

Science.gov (United States)

Nugraha Pratama, Sendi; Kurniawan, Yudhi; Muhammady, Shibghatullah; Takase, Kouichi; Darma, Yudi

2018-03-01

We study the formation of nickel nanowires (Ni NWs) grown in porous anodic aluminium oxide (AAO) template by the electrochemical deposition technique. Here, the initial AAO template was grown by anodization of aluminium substrate in sulphuric acid solution. The cross-section, crystal structure, and magnetic properties of Ni NWs system were characterized by field-emission SEM, XRD, and SQUID. As a result, the highly-ordered Ni NWs are observed with the uniform diameter of 27 nm and the length from 31 to 163 nm. Based on XRD spectra analysis, Ni NWs have the face-centered cubic structure with the lattice parameter of 0.35 nm and average crystallite size of 17.19 nm. From SQUID measurement at room temperature, by maintaining the magnetic field perpendicular to Ni NWs axis, the magnetic hysteresis of Ni NWs system show the strong ferromagnetism with the coercivity and remanence ratio of ∼148 Oe and ∼0.23, respectively. The magnetic properties are also calculated by means of generalized gradient approximation methods. From the calculation result, we show that the ferromagnetism behavior comes from Ni NWs without any contribution from AAO template or the substrate. This study opens the potential application of Ni NWs system for novel functional magnetic devices.

Microstructural and electrochemical characterization of Ni/Ti/sub 2/N composite coating for sintered NdFeB permanent magnets

International Nuclear Information System (INIS)

Ali, A.; Ahmad, A.; Deen, K.M.; Ahmad, R.

2009-01-01

Sintered NdFeB magnets have complex microstructure that makes them susceptible to corrosion in humid or moist environments. The paper presents the anticorrosion characteristics of a novel Ni/Ti/sub 2/N composite coating applied through electrodeposition and cathodic arc physical vapour deposition (CAPVD) to sintered NdFeB permanent magnets. The performance of composite coating was evaluated in simulated marine environment with the help of dc polarization techniques. The rate of coating degradation was also determined by employing ac electrochemical impedance spectroscopy (EIS). The coating morphology and surface chemistry were studied with scanning electron microscope (SEM). X-ray diffraction (XRD) was used for identification of component phases in the coating-substrate system. The results showed that the composite coating provided an adequately improved corrosion protection to the sintered NdFeB magnets in the simulated marine environment compared to the earlier reported ceramic and metallic coatings. The composite coating did not damage the magnetic properties of coating-substrate system that remained at par with the ceramic and nickel coating having copper interlayer. (author)

Electrochemical approach for acute myocardial infarction diagnosis based on direct antibodies-free analysis of human blood plasma.

Science.gov (United States)

Suprun, Elena V; Saveliev, Anatoly A; Evtugyn, Gennady A; Lisitsa, Alexander V; Bulko, Tatiana V; Shumyantseva, Victoria V; Archakov, Alexander I

2012-03-15

A novel direct antibodies-free electrochemical approach for acute myocardial infarction (AMI) diagnosis has been developed. For this purpose, a combination of the electrochemical assay of plasma samples with chemometrics was proposed. Screen printed carbon electrodes modified with didodecyldimethylammonium bromide were used for plasma charactrerization by cyclic (CV) and square wave voltammetry and square wave (SWV) voltammetry. It was shown that the cathodic peak in voltammograms at about -250 mV vs. Ag/AgCl can be associated with AMI. In parallel tests, cardiac myoglobin and troponin I, the AMI biomarkers, were determined in each sample by RAMP immunoassay. The applicability of the electrochemical testing for AMI diagnostics was confirmed by statistical methods: generalized linear model (GLM), linear discriminant analysis (LDA) and quadratic discriminant analysis (QDA), artificial neural net (multi-layer perception, MLP), and support vector machine (SVM), all of which were created to obtain the "True-False" distribution prediction where "True" and "False" are, respectively, positive and negative decision about an illness event. Copyright © 2011 Elsevier B.V. All rights reserved.

Synthesis, structure, magnetic, electrical and electrochemical properties of Al, Cu and Mg doped MnO{sub 2}

Energy Technology Data Exchange (ETDEWEB)

Hashem, Ahmed M., E-mail: ahmedh242@yahoo.com [National Research Centre, Inorganic Chemistry Department, Behoes St., Dokki, Cairo (Egypt); Institute for Complex Materials, IFW Dresden, Helmholtzstr. 20, D-01069 Dresden (Germany); Abuzeid, Hanaa M. [National Research Centre, Inorganic Chemistry Department, Behoes St., Dokki, Cairo (Egypt); Narayanan, N. [Institute for Complex Materials, IFW Dresden, Helmholtzstr. 20, D-01069 Dresden (Germany); Ehrenberg, Helmut [Institute for Complex Materials, IFW Dresden, Helmholtzstr. 20, D-01069 Dresden (Germany); Materials Science, Technische Universitaet Darmstadt, Petersenstr. 23, D-64287 Darmstadt (Germany); Julien, C.M. [Universite Pierre et Marie Curie, Physicochimie des Electrolytes, Colloides et Sciences Analytiques (PECSA), 4 place Jussieu, 75005 Paris (France)

2011-10-17

Highlights: {yields} Al, Mg and Cu doped MnO{sub 2} as cathode in Li-ion batteries. {yields} Pure phase MnO{sub 2} for virgin and doped MnO{sub 2} were obtained. {yields} Doping elements improve the electrical conductivity of MnO{sub 2}. {yields} Electrochemical behaviour of MnO{sub 2} improved after doping by Al, Mg and Cu. - Abstract: Pure and doped manganese dioxides were prepared by wet-chemical method using fumaric acid and potassium permanganate as raw materials. X-ray diffraction patterns show that pure and Al, Cu and Mg doped manganese dioxides (d-MnO{sub 2}) crystallized in the cryptomelane-MnO{sub 2} structure. Thermal analysis show that, with the assistance of potassium ions inside the 2 x 2 tunnel, the presence of Al, Cu and Mg doping elements increases the thermal stability of d-MnO{sub 2}. The electrical conductivity of d-MnO{sub 2} increases in comparison with pure MnO{sub 2}, while Al-doped MnO{sub 2} exhibits the lower resistivity. As shown in the magnetic measurements, the value of the experimental effective magnetic moment of Mn ions decreases with introduction of dopants, which is attributed to the presence of a mixed valency of high-spin state Mn{sup 4+}/Mn{sup 3+}. Doped MnO{sub 2} materials show good capacity retention in comparison with virgin MnO{sub 2}. Al-doped MnO{sub 2} shows the best electrochemical results in terms of capacity retention and recharge efficiency.

Heterogeneous time-resolved electrochemiluminoimmunoassay of thyroid stimulating hormone with magnetic beads at oxide-covered aluminum electrode

Energy Technology Data Exchange (ETDEWEB)

Ala-Kleme, Timo, E-mail: timo.ala-kleme@utu.fi

2017-06-15

The heterogeneous immunoassay of thyroid stimulating hormone (TSH) was detected by the time-resolved cathodic electrochemiluminescence immunoassay (tr-CECLIA) method using magnetic beads as a mobile support. The magnetic beads coated with sandwich type TSH immunoassay were captured with a magnet for washing and detection processes. The time-resolved cathodic electrochemiluminescence (tr-CECL) signal of Tb(III) chelate label was generated by cathodic pulse polarization in the aluminum working electrode and platinum counter electrode system. The detection method causes injection of high energy electrons into the aqueous solution near the aluminum electrode and creates rigid simultaneous oxidative and reductive conditions that excitate the Tb(III) chelate used as a label luminophore in the heterogeneous sandwich type immunoassay of TSH in the surface of the magnetic beads. The limit of detection of the method was about 50 mIU L{sup −1}. The precision of it was noticed to be good; the coefficient variation percentage was realized to be lower than 10 %. Unfortunately the limit of detection is not good enough for determination of analyte levels of very low concentrations for instance TSH in body fluids. The possible application areas of the method are in highly sophisticated micro or nano fluidic detection and sensor systems where the aspiration level of the analyte detection limit is not very high and the mobility and manageability and the large surface area of the magnetic beads can be utilized efficiently in separating, washing, moving, coating and detecting processes. In the case of tr-CECLIA the presented method makes possible to use multipurpose working electrodes instead of disposable ones.

Development of nanobody-based flow injection chemiluminescence immunoassay for sensitive detection of human prealbumin.

Science.gov (United States)

Ma, Lei; Sun, Yanyan; Kang, Xuejun; Wan, Yakun

2014-11-15

Nanobodies, derived from camelid heavy-chain antibodies, have novel and impactful applications in clinical diagnostics. Our objective is to develop a nanobody-based chemiluminescence immunoassay for sensitive detection of human prealbumin (PA). In this context, a phage display nanobody library is constructed via immunizing dromedary camel with human prealbumin. Three nanobodies have been identified by five successive bio-panning steps. Based on their high expression level and good affinity, two out of three are chosen for further study. Magnetic beads (MBs) were functionalized with PEI by acylamide bond formed between the carboxyl group on the surface of the MB. Then, an anti-PA nanobody (Nb1) can be effectively immobilized onto the surface of the functionalized MB using glutaradehyde as the link. The modified MBs with Nb1 can specifically capture the target PA and reacted with silica nanoparticles with co-immobilized HRP and anti-PA nanobody (Nb2). The concentration of PA was detected by flow injection chemiluminescence. When using MB/PEI as the carrier of anti-PA Nb1, the CL signal significantly increased to 4-fold compared with the signal using MB without PEI modification. The CL signal was further amplified to 5-fold when Si/Nb2 was used as the signal probe. Under optimized conditions, the present immunoassay exhibited a wide quantitative range from 0.05 to 1000 μg L(-1) with a detection limit of 0.01 μg L(-1). The sensitivity of the proposed immunoassay offers great promises in providing a sensitive, specific, time saving, and potential method for detecting PA in clinical settings. Copyright © 2014 Elsevier B.V. All rights reserved.

Micromotor-based lab-on-chip immunoassays

Science.gov (United States)

García, Miguel; Orozco, Jahir; Guix, Maria; Gao, Wei; Sattayasamitsathit, Sirilak; Escarpa, Alberto; Merkoçi, Arben; Wang, Joseph

2013-01-01

Here we describe the first example of using self-propelled antibody-functionalized synthetic catalytic microengines for capturing and transporting target proteins between the different reservoirs of a lab-on-a-chip (LOC) device. A new catalytic polymer/Ni/Pt microtube engine, containing carboxy moieties on its mixed poly(3,4-ethylenedioxythiophene) (PEDOT)/COOH-PEDOT polymeric outermost layer, is further functionalized with the antibody receptor to selectively recognize and capture the target protein. The new motor-based microchip immunoassay operations are carried out without any bulk fluid flow, replacing the common washing steps in antibody-based protein bioassays with the active transport of the captured protein throughout the different reservoirs, where each step of the immunoassay takes place. A first microchip format involving an `on-the-fly' double-antibody sandwich assay (DASA) is used for demonstrating the selective capture of the target protein, in the presence of excess of non-target proteins. A secondary antibody tagged with a polymeric-sphere tracer allows the direct visualization of the binding events. In a second approach the immuno-nanomotor captures and transports the microsphere-tagged antigen through a microchannel network. An anti-protein-A modified microengine is finally used to demonstrate the selective capture, transport and convenient label-free optical detection of a Staphylococcus aureus target bacteria (containing proteinA in its cell wall) in the presence of a large excess of non-target (Saccharomyces cerevisiae) cells. The resulting nanomotor-based microchip immunoassay offers considerable potential for diverse applications in clinical diagnostics, environmental and security monitoring fields.Here we describe the first example of using self-propelled antibody-functionalized synthetic catalytic microengines for capturing and transporting target proteins between the different reservoirs of a lab-on-a-chip (LOC) device. A new catalytic

Determining total thyroxine with the aid of the homogeneous enzyme immunoassay

International Nuclear Information System (INIS)

Vogt, W.

1978-01-01

Total thyrozine values obtained with the aid of the homogeneous enzyme immunoassay are compared with those delivered by the RIA and the efficiency of the EMIT technique is evaluated. Some results obtained via Enzymun-T4, a heterologous enzyme immunoassay, are also given. (VJ) 891 VJ [de

A disposable electrochemical immunosensor based on carbon screen-printed electrodes for the detection of prostate specific antigen.

Science.gov (United States)

Yan, Mei; Zang, Dejin; Ge, Shenguang; Ge, Lei; Yu, Jinghua

2012-01-01

A novel screen-printed electrode (SPEs) on sheets of vegetable parchment was prepared. The obtained SPEs were stable, convenient, inexpensive and suitable for large-area screen-printing. With these SPEs, we explored the fabrication of a novel, disposable and highly sensitive electro-analytical immunosensor using graphene nanosheets (GS) and horseradish peroxidase (HRP)-labeled signal antibody functionalized with gold nanoparticles (HRP-Ab(2)/Au NPs). GS was used to increase the conductivity and stability of this immunosensor due to its fast electron transportation and good biocompatibility. Au NPs could not only provide a large surface area for the immobilization of HRP-Ab(2) but also enhance the electroreduction between HRP and H(2)O(2) to amplify the electrochemical signal on the sandwich immuno-complexes modified SPEs. The proposed SPEs were characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and electrochemical methods involving cyclic voltammetry (CV), and electrochemical impedence method. Using prostate specific antigen (PSA) as a model analyte, this immunosensor showed a wide linear range over 6 orders of magnitude with the minimum value down to 2 pg mL(-1). In addition, this immunosensor could avoid the need of deoxygenation for the electrochemical immunoassay. Thus, it provided a promising potential in clinical applications. Copyright © 2012 Elsevier B.V. All rights reserved.

Enhancing graphene/CNT based electrochemical detection using magneto-nanobioprobes

OpenAIRE

sprotocols

2015-01-01

Authors: Priyanka Sharma, V Bhalla, E Senthil Prasad, V Dravid, G Shekhawat & C. Raman Suri ### Abstract This protocol describes an optimized signal amplification strategy to develop an ultra-sensitive magneto-electrochemical biosensing platform. The new protocol combines the advantages of carbon nanotube (CNT) and reduced graphene oxide (rGO) together with electrochemical bursting of magnetic nanoparticles. The method involves synthesis of gold-iron (Au/Fe) nano-structures function...

Highly sensitive immunoassay based on E. coli with autodisplayed Z-domain

International Nuclear Information System (INIS)

Jose, Joachim; Park, Min; Pyun, Jae-Chul

2010-01-01

The Z-domain of protein A has been known to bind specifically to the F c region of antibodies (IgGs). In this work, the Z-domain of protein A was expressed on the outer membrane of Escherichia coli by using 'Autodisplay' technology as a fusion protein of autotransport domain. The E. coli with autodisplayed Z-domain was applied to the sandwich-type immunoassay as a solid-support of detection-antibodies against a target analyte. For the feasibility demonstration of the E. coli based immunoassay, C-reactive protein (CRP) assay was carried out by using E. coli with autodisplayed Z-domain. The limit of detection (LOD) and binding capacity of the E. coli based immunoassay were estimated to be far more sensitive than the conventional ELISA. Such a far higher sensitivity of E. coli based immunoassay than conventional ELISA was explained by the orientation control of immobilized antibodies and the mobility of E. coli in assay matrix. From the test results of 45 rheumatoid arthritis (RA) patients' serum and 15 healthy samples, a cut-off value was established to have optimal sensitivity and selectivity values for RA. The CRP test result of each individual sample was compared with ELISA which is the reference method for RA diagnosis. From this work, the E. coli with Z-domain was proved to be feasible for the medical diagnosis based on sandwich-type immunoassay.

Magnetic field-assisted synthesis of wire-like Co3O4 nanostructures: Electrochemical and photocatalytic studies

International Nuclear Information System (INIS)

Zhao, Xiubin; Pang, Zhanwen; Wu, Mingzai; Liu, Xiansong; Zhang, Hui; Ma, Yongqing; Sun, Zhaoqi; Zhang, Lide; Chen, Xiaoshuang

2013-01-01

Graphical abstract: Schematic illustration for the magnetic field-assisted growth of wire-like Co 3 O 4 nanostructure. Display Omitted Highlights: ► Co 3 O 4 nanowires are prepared by magnetic field hydrothermal reduction and annealing. ► These Co 3 O 4 nanowires possess enhanced capacitance. ► The Co 3 O 4 nanowires have a good photocatalytic activity for methyl orange. -- Abstract: Wire-like Co 3 O 4 nanostructures were prepared by the combination of magnetic field-assisted hydrothermal reduction of cobalt ions and the subsequent ambient annealing at 500 °C. X-ray diffraction (XRD) and scanning electron microscopy (SEM) were used to characterize the structure and morphological evolution of the products. The results show that the wire-like nanostructures possess diameters about 250 nm and lengths over 10 μm. The possible formation mechanism of the wire-like Co 3 O 4 nanostructures is also proposed based on the SEM results. Galvanostatic methods were used to characterize the electrochemical properties. The measurements indicate that the wire-like Co 3 O 4 nanostructures show larger discharge and charge capacities than that of spherical Co 3 O 4 nanoparticles prepared in the absence of magnetic field. In addition, the photocatalytic activity of the products was investigated by measuring the photodegradation of methyl orange solution under ultraviolet radiation, which shows that both the wire-like and spherical products have a good photocatalytic activity.

Multiplex Immunoassay Profiling of Hormones Involved in Metabolic Regulation.

Science.gov (United States)

Stephen, Laurie; Guest, Paul C

2018-01-01

Multiplex immunoassays are used for rapid profiling of biomarker proteins and small molecules in biological fluids. The advantages over single immunoassays include lower sample consumption, cost, and labor. This chapter details a protocol to develop a 5-plex assay for glucagon-like peptide 1, growth hormone, insulin, leptin, and thyroid-stimulating hormone on the Luminex ® platform. The results of the analysis of insulin in normal control subjects are given due to the important role of this hormone in nutritional programming diseases.

Francisella tularensis detection using magnetic labels and a magnetic biosensor based on frequency mixing

International Nuclear Information System (INIS)

Meyer, Martin H.F.; Krause, Hans-Joachim; Hartmann, Markus; Miethe, Peter; Oster, Juergen; Keusgen, Michael

2007-01-01

A biosensor that uses resonant coils with a special frequency-mixing technique and magnetic beads as detectable labels has been established for the detection of Francisella tularensis, the causative agent for tularemia. The detection principle is based on a sandwich immunoassay using an anti-Ft antibody for immunofiltration immobilized to ABICAP[reg] polyethylene filters, and biotinylated with streptavidin-coated magnetic beads as labels. The linear detection range of this biosensor was found to be 10 4 -10 6 cfu F. tularensis lipopolysaccharide (LPS) per ml. Tested sample matrices were physiological PBS buffer and rabbit serum

Potential applications of immunoassays in studies of flatfish recruitment

Science.gov (United States)

Feller, Robert J.

The fisheries recruitment-stock problem, a lack of correlation between measures of reproductive output of the parent stock and recruitment to the fishery, has several potential biotic and abiotic causes. Immunoassays may be useful in examining several aspects of this and several other problems in flatfish ecology: stock identification, parasitism and disease, and trophic interactions. Given stage-specific antisera capable of recognozing antigenic moieties of, for instance, eggs, larvae, or newly-settled juveniles, it is possible to screen stomach contents of many putative predators ( e.g., shrimp or crabs) rapidly for the presence and amounts of platfish prey. This trophic application of immunological methods has great promise for measuring loss of potential recruits to predation. All immunoassays are limited by the quality of antisera used and the researcher's ability to interpret quantitative data in an ecologically meaningful way. Key references for applications of immunoassays in fish-related questions are provided with recommendations for their utilization.

Negative interference by rheumatoid factor in alpha-fetoprotein chemiluminescent microparticle immunoassay.

Science.gov (United States)

Wang, Hui; Bi, Xiaohui; Xu, Lei; Li, Yirong

2017-01-01

Background Rheumatoid factor causes positive interference in multiple immunoassays. Recently, negative interference has also been found in immunoassays in the presence of rheumatoid factor. The chemiluminescent microparticle immunoassay is widely used to determine serum alpha-fetoprotein. However, it is not clear whether the presence of rheumatoid factor in the serum causes interference in the chemiluminescent microparticle immunoassay of alpha-fetoprotein. Methods Serum alpha-fetoprotein was determined using the ARCHITECT alpha-fetoprotein assay. The estimation of alpha-fetoprotein recovery was carried out in samples prepared by diluting high-concentration alpha-fetoprotein serum with rheumatoid factor-positive or rheumatoid factor-negative serum. Paramagnetic microparticles coated with hepatitis B surface antigen-anti-HBs complexes were used to remove rheumatoid factor from the serum. Results The average recovery of alpha-fetoprotein was 88.4% and 93.8% in the rheumatoid factor-positive and rheumatoid factor-negative serum samples, respectively. The recovery of alpha-fetoprotein was significantly lower in the rheumatoid factor-positive serum samples than in the rheumatoid factor-negative serum samples. In two of five rheumatoid factor-positive samples, a large difference was found (9.8%) between the average alpha-fetoprotein recoveries in the serially diluted and initial recoveries. Fourteen rheumatoid factor-positive serum samples were pretreated with hepatitis B surface antigen-anti-HBs complex-coated paramagnetic microparticles. The alpha-fetoprotein concentrations measured in the pretreated samples increased significantly. Conclusions It was concluded that the alpha-fetoprotein chemiluminescent microparticle immunoassay is susceptible to interference by rheumatoid factor, leading to significantly lower results. Eliminating the incidence of negative interference from rheumatoid factor should be an important goal for immunoassay providers. In the meantime

Development of a novel ultrasensitive enzyme immunoassay for human glutamic acid decarboxylase 65 antibody.

Science.gov (United States)

Numata, Satoshi; Katakami, Hideki; Inoue, Shinobu; Sawada, Hirotake; Hashida, Seiichi

2016-07-01

We developed a novel, ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) for determination of glutamic acid decarboxylase autoantibody concentrations in serum samples from patients with type 2 diabetes. We developed an immune complex transfer enzyme immunoassay for glutamic acid decarboxylase autoantibody and measured glutamic acid decarboxylase autoantibody from 22 patients with type 1 diabetes, 29 patients with type 2 diabetes, and 32 healthy controls. A conventional ELISA kit identified 10 patients with type 1 diabetes and one patient with type 2 diabetes as glutamic acid decarboxylase autoantibody positive, whereas 15 patients with type 1 diabetes and six patients with type 2 diabetes were identified as glutamic acid decarboxylase autoantibody positive using immune complex transfer enzyme immunoassay. Immune complex transfer enzyme immunoassay is a highly sensitive and specific assay for glutamic acid decarboxylase autoantibody and might be clinically useful for diabetic onset prediction and early diagnosis. © The Author(s) 2016.

Heparin interferes with the radioenzymatic and homogeneous enzyme immunoassays for aminoglycosides

International Nuclear Information System (INIS)

Krogstad, D.J.; Granich, G.G.; Murray, P.R.; Pfaller, M.A.; Valdes, R.

1981-01-01

Heparin interferes with measurement of aminoglycosides in serum by biological, radioenzymatic, and homogeneous enzyme immunoassay techniques, but not with radioimmunoassay. At concentrations greater than or equal to 10 5 and greater than or equal to 3 X 10 6 USP units/L, respectively, it interferes with the radioenzymatic assay by inhibiting the gentamicin 3-acetyltransferase and kanamycin 6'-acetyltransferase enzymes used in the assay. It interferes with the homogeneous enzyme immunoassays for gentamicin and tobramycin (at concentrations greater than or equal to 10 5 and greater than or equal to10 4 USP units/L, respectively), but not with the commercially available homogeneous enzyme immunoassays for other drugs. Heparin interference with the homogeneous enzyme immunoassay for aminoglycosides requires both the heparin polyanion and glucose-6-phosphate dehydrogenase bound to a cationic aminoglycoside. This interference can be reproduced with dextran sulfate (but not dextran), and does not occur with free enzyme (glucose-6-phosphate dehydrogenase) alone. Heparin interference with these two assays and at concentrations that may be present in intravenous infusions or in seriously underfilled blood-collection tubes is described

Magnetic effects in electrochemistry

Directory of Open Access Journals (Sweden)

NEBOJSA D. NIKOLIC

2005-05-01

Full Text Available The effect of imposed magnetic fields onto the electrodeposition of magnetic (nickel and non – magnetic (copper metals was analysed. Also, magnetic properties of electrochemically obtained nanocontacts were examined. An effort to establish a possible correlation between the morphologies of the nanocontacts and the effect of the very large ballistic magnetoresistance (BMR effect was made.

Sensitivity-Enhancement of FRET Immunoassays by Multiple-Antibody Conjugation on Quantum Dots.

Science.gov (United States)

Annio, Giacomo; Jennings, Travis; Tagit, Oya; Hildebrandt, Niko

2018-05-23

Quantum dots (QDs) are not only advantageous for color-tuning, improved brightness, and high stability, but their nanoparticle surfaces also allow for the attachment of many biomolecules. Because IgG antibodies (ABs) are in the same size range of biocompatible QDs and the AB orientation after conjugation to the QD is often random, it is difficult to predict if few or many ABs per QD will lead to an efficient AB-QD conjugate. This is particularly true for homogeneous Förster resonance energy transfer (FRET) sandwich immunoassays, for which the ABs on the QD must bind a biomarker that needs to bind a second AB-FRET-conjugate. Here, we investigate the performance of Tb-to-QD FRET immunoassays against total prostate specific antigen (TPSA) by changing the number of ABs per QD while leaving all the other assay components unchanged. We first characterize the AB-QD conjugation by various spectroscopic, microscopic, and chromatographic techniques and then quantify the TPSA immunoassay performance regarding sensitivity, limit of detection, and dynamic range. Our results show that an increasing conjugation ratio leads to significantly enhanced FRET immunoassays. These findings will be highly important for developing QD-based immunoassays in which the concentrations of both ABs and QDs can significantly influence the assay performance.

Development of an Electrochemical Biosensor for the Detection of Aflatoxin M1 in Milk

Directory of Open Access Journals (Sweden)

Jean-Louis Marty

2010-10-01

Full Text Available We have developed an electrochemical immunosensor for the detection of ultratrace amounts of aflatoxin M1 (AFM1 in food products. The sensor was based on a competitive immunoassay using horseradish peroxidase (HRP as a tag. Magnetic nanoparticles coated with antibody (anti-AFM1 were used to separate the bound and unbound fractions. The samples containing AFM1 were incubated with a fixed amount of antibody and tracer [AFM1 linked to HRP (conjugate] until the system reached equilibrium. Competition occurs between the antigen (AFM1 and the conjugate for the antibody. Then, the mixture was deposited on the surface of screen-printed carbon electrodes, and the mediator [5-methylphenazinium methyl sulphate (MPMS] was added. The enzymatic response was measured amperometrically. A standard range (0, 0.005, 0.01, 0.025, 0.05, 0.1, 0.25, 0.3, 0.4 and 0.5 ppb of AFM1-contaminated milk from the ELISA kit was used to obtain a standard curve for AFM1. To test the detection sensitivity of our sensor, samples of commercial milk were supplemented at 0.01, 0.025, 0.05 or 0.1 ppb with AFM1. Our immunosensor has a low detection limit (0.01 ppb, which is under the recommended level of AFM1 [0.05 µg L-1 (ppb], and has good reproducibility.

Interference in immunoassay

International Nuclear Information System (INIS)

Chapman, R.S.

1998-01-01

Interfering factors are evident in both limited reagent (radioimmunoassay) and excess reagent (immunometric assay) technologies and should be suspected whenever there is a discrepancy between analytical results and clinical findings in the investigation of particular diseases. The overall effect of interference in immunoassay is analytical bias in result, either positive or negative of variable magnitude. The interference maybe caused by a wide spectrum of factors from poor sample collection and handling to physiological factors e.g. lipaemia, heparin treatment, binding protein abnormalities, autoimmunity and drug treatments. The range of interfering factors is extensive and difficult to discuss effectively in a short review

Sensitive electrochemical immunosensor for α-fetoprotein based on graphene/SnO2/Au nanocomposite.

Science.gov (United States)

Liu, Junfeng; Lin, Guanhua; Xiao, Can; Xue, Ying; Yang, Ankang; Ren, Hongxuan; Lu, Wensheng; Zhao, Hong; Li, Xiangjun; Yuan, Zhuobin

2015-09-15

A label-free electrochemical immunosensor for sensitive detection of α-fetoprotein (AFP) was developed based on graphene/SnO2/Au nanocomposite. The graphene/SnO2/Au nanocomposite modified glassy carbon electrode was used to immobilize α-fetoprotein antibody (anti-AFP) and to construct the immunosensor. Results demonstrated that the peak currents of [Ru(NH3)6](3+) decreased due to the interaction between antibody and antigen on the modified electrode. Thus, a label-free immunosensor for the detection of AFP was realized by monitoring the peak current change of [Ru(NH3)6](3+). The factors influencing the performance of the immunosensor were investigated in details. Under optimal conditions, the peak currents obtained by DPV decreased linearly with the increasing AFP concentrations in the range from 0.02 to 50 ng mL(-1) with a linear coefficient of 0.9959. This electrochemical immunoassay has a low detection limit of 0.01 ng mL(-1) (S/N=3) and was successfully applied to the determination of AFP in serum samples. Copyright © 2015 Elsevier B.V. All rights reserved.

Procedures for Sensitive Immunoassay

Energy Technology Data Exchange (ETDEWEB)

Givol, D. [Department of Chemical Immunology, Weizmann Institute of Science, Rehovot (Israel)

1970-02-15

Sensitive immunoassay methods should be applied to small molecules of biological importance, which are non-immunogenic by themselves, such as small peptide hormones (e.g. bradykinin), plant hormones (e.g. indoleacetic acid), nucleotides and other small molecules. Methods of binding these small molecules, as haptens, to immunogenic carriers by various cross-linking agents are described (dicyclohexylcarbodiimide, tolylene-diisocyanate and glutaraldehyde), and the considerations involved in relation to the methods of binding and the specificity of the antibodies formed are discussed. Some uses of antibody bound to bromoacetyl cellulose as an immuno adsorbent convenient for assay of immunoglobulins are described. Finally, the sensitive immunoassay method of chemically modified phage is described. This includes methods of binding small molecules (such as the dinitrophenyl group, penicillin, indoleacetic acid) or proteins (such as insulin, immunoglobulins) to phages. Methods of direct chemical conjugation, or an indirect binding via anti-phage Fab, are described. The phage inactivation method by direct plating and its modifications (such as decision technique and complex inactivation) are compared with the more simple end-point titration method. The inhibition of phage inactivation has some advantages as it does not require radioactive material, or expensive radioactive counters, and avoids the need for separation between bound and unbound antigen. Hence, if developed, it could be used as an alternative to radioimmunoassay. (author)

Francisella tularensis detection using magnetic labels and a magnetic biosensor based on frequency mixing

Energy Technology Data Exchange (ETDEWEB)

Meyer, Martin H.F. [Institute for Pharmaceutical Chemistry, Philipps-Universitaet Marburg (Germany); Krause, Hans-Joachim [Institute of Bio-and Nanosystems (IBN-2), Research Center Juelich (Germany); Hartmann, Markus [Institute for Pharmaceutical Chemistry, Philipps-Universitaet Marburg (Germany); Miethe, Peter [SENOVA GmbH, Jena (Germany); Oster, Juergen [chemagen GmbH, Baesweiler (Germany); Keusgen, Michael [Institute for Pharmaceutical Chemistry, Philipps-Universitaet Marburg (Germany)]. E-mail: Keusgen@staff.uni-marburg.de

2007-04-15

A biosensor that uses resonant coils with a special frequency-mixing technique and magnetic beads as detectable labels has been established for the detection of Francisella tularensis, the causative agent for tularemia. The detection principle is based on a sandwich immunoassay using an anti-Ft antibody for immunofiltration immobilized to ABICAP[reg] polyethylene filters, and biotinylated with streptavidin-coated magnetic beads as labels. The linear detection range of this biosensor was found to be 10{sup 4}-10{sup 6} cfu F. tularensis lipopolysaccharide (LPS) per ml. Tested sample matrices were physiological PBS buffer and rabbit serum.

Yoctomole electrochemical genosensing of Ebola virus cDNA by rolling circle and circle to circle amplification.

Science.gov (United States)

Carinelli, S; Kühnemund, M; Nilsson, M; Pividori, M I

2017-07-15

This work addresses the design of an Ebola diagnostic test involving a simple, rapid, specific and highly sensitive procedure based on isothermal amplification on magnetic particles with electrochemical readout. Ebola padlock probes were designed to detect a specific L-gene sequence present in the five most common Ebola species. Ebola cDNA was amplified by rolling circle amplification (RCA) on magnetic particles. Further re-amplification was performed by circle-to-circle amplification (C2CA) and the products were detected in a double-tagging approach using a biotinylated capture probe for immobilization on magnetic particles and a readout probe for electrochemical detection by square-wave voltammetry on commercial screen-printed electrodes. The electrochemical genosensor was able to detect as low as 200 ymol, corresponding to 120 cDNA molecules of L-gene Ebola virus with a limit of detection of 33 cDNA molecules. The isothermal double-amplification procedure by C2CA combined with the electrochemical readout and the magnetic actuation enables the high sensitivity, resulting in a rapid, inexpensive, robust and user-friendly sensing strategy that offers a promising approach for the primary care in low resource settings, especially in less developed countries. Copyright © 2016 Elsevier B.V. All rights reserved.

Abbott prism: a multichannel heterogeneous chemiluminescence immunoassay analyzer.

Science.gov (United States)

Khalil, O S; Zurek, T F; Tryba, J; Hanna, C F; Hollar, R; Pepe, C; Genger, K; Brentz, C; Murphy, B; Abunimeh, N

1991-09-01

We describe a multichannel heterogeneous immunoassay analyzer in which a sample is split between disposable reaction trays in a group of linear tracks. The system's pipettor uses noninvasive sensing of the sample volume and disposable pipet tips. Each assay track has (a) a conveyor belt for moving reaction trays to predetermined functional stations, (b) temperature-controlled tunnels, (c) noncontact transfer of the reaction mixture between incubation and detection wells, and (d) single-photon counting to detect a chemiluminescence (CL) signal from the captured immunochemical product. A novel disposable reaction tray, with separate reaction and detection wells and self-contained fluid removal, is used in conjunction with the transfer device on the track to produce a carryover-free system. The linear immunoassay track has nine predetermined positions for performing individual assay steps. Assay step sequence and timing is selected by changing the location of the assay modules between these predetermined positions. The assay methodology, a combination of microparticle capture and direct detection of a CL signal on a porous matrix, offers excellent sensitivity, specificity, and ease of automation. Immunoassay configurations have been tested for hepatitis B surface antigen and for antibodies to hepatitis B core antigen, hepatitis C virus, human immunodeficiency virus I and II, and human T-cell leukemia virus I and II.

Immunoassay for determination of trilobolide

Czech Academy of Sciences Publication Activity Database

Huml, L.; Jurášek, M.; Mikšátková, P.; Zimmermann, T.; Tomanová, P.; Buděšínský, Miloš; Rottnerová, Z.; Šimková, M.; Harmatha, Juraj; Kmoníčková, Eva; Lapčík, O.; Drašar, P. B.

2017-01-01

Roč. 117, Jan (2017), s. 105-111 ISSN 0039-128X. [Conference on Isoprenoids /23./. Minsk, 04.09.2016-07.09.2016] Institutional support: RVO:61388963 ; RVO:68378041 Keywords : trilobolide * avidin-biotin * ELISA * Laser trilobum * synthesis * immunoassay Subject RIV: CE - Biochemistry; FR - Pharmacology ; Medidal Chemistry (UEM-P) OBOR OECD: Biochemical research methods; Pharmacology and pharmacy (UEM-P) Impact factor: 2.282, year: 2016

Development of immunoassays for detecting clothianidin residue in agricultural products.

Science.gov (United States)

Li, Ming; Sheng, Enze; Cong, Lujing; Wang, Minghua

2013-04-17

Two enzyme-linked immunosorbent assays (ELISAs) based on polyclonal antibodies (PcAbs) for clothianidin are described: colorimetric detection format (ELISA) and pattern of chemiluminescent assay (CLEIA). Clothianidin hapten was synthesized and conjugated to bovine serum albumin (BSA) and ovalbumin (OVA) to produce immunogen and coating antigen. Anticlothianidin PcAbs were obtained from immunized New Zealand white rabbits. Under optimal conditions, the half-maximal inhibition concentration (IC₅₀) and the limit of detection (LOD, IC₂₀) of clothianidin were 0.046 and 0.0028 mg/L for the ELISA and 0.015 and 0.0014 mg/L for the CLEIA, respectively. There were no obvious cross-reactivities of the antibodies with its analogues except for dinotefuran. Recoveries of 76.4-116.4% for the immunoassays were achieved from spiked samples. The results of immunoassays for the spiked and authentic samples were largely consistent with gas chromatography. Therefore, the proposed immunoassays would be convenient and satisfactory analytical methods for the monitoring of clothianidin in agricultural products.

AN ENVIRONMENTAL TECHNOLOGY VERIFICATION (ETV) TESTING OF THREE IMMUNOASSAY TEST KITS FOR ANTHRAX, BOTULINUM TOXIN AND RICIN

Science.gov (United States)

Immunoassay test kits are based on immunoassay methods, where specific antibodies are used to detect and measure the contaminants of interest. Immunoassay test kits rely on the reaction of a contaminant or antigen with a selective antibody to give a product that can be measures....

Rapid electrochemical quantification of Salmonella Pullorum and Salmonella Gallinarum based on glucose oxidase and antibody-modified silica nanoparticles.

Science.gov (United States)

Luo, Yiheng; Dou, Wenchao; Zhao, Guangying

2017-07-01

In this article, a facile and sensitive electrochemical method for quantification of Salmonella Pullorum and Salmonella Gallinarum (S. Pullorum and S. Gallinarum) was established by monitoring glucose consumption with a personal glucose meter (PGM). Antibody-functionalized magnetic nanoparticles (IgG-MNPs) were used to capture and enrich S. Pullorum and S. Gallinarum, and IgG-MNPs-S. Pullorum and IgG-MNPs-S. Gallinarum complexes were magnetically separated from a sample using a permanent magnet. The trace tag was prepared by loading polyclonal antibodies and high-content glucose oxidase on amino-functionalized silica nanoparticles (IgG-SiNPs-GOx). With a sandwich-type immunoassay format, IgG-SiNPs-GOx were added into the above mixture solution and conjugated to the complexes, forming sandwich composites IgG-MNPs/S. Pullorum and S. Gallinarum/IgG-SiNPs-GOx. The above sandwich composites were dispersed in glucose solution. Before and after the hydrolysis of glucose, the concentration of glucose was measured using PGM. Under optimal conditions, a linear relationship between the decrease of glucose concentration and the logarithm of S. Pullorum and S. Gallinarum concentration was obtained in the concentration range from 1.27 × 10 2 to 1.27 × 10 5  CFU mL -1 , with a detection limit of 7.2 × 10 1  CFU mL -1 (S/N = 3). This study provides a portable, low-cost, and quantitative analytical method for bacteria detection; thus, it has a great potential in the prevention of disease caused by S. Pullorum and S. Gallinarum in poultry. Graphical abstract A schematic illustration of the fabrication process of IgG-SiNPs-GOD nanomaterials (A) and IgG-MNPs (B) and experimental procedure of detection of S. Pullorum and S. Gallinarum using GOD-functionalized silica nanospheres as trace tags based on PGM (C).

Magnetic actuator for the control and mixing of magnetic bead-based reactions on-chip.

Science.gov (United States)

Berenguel-Alonso, Miguel; Granados, Xavier; Faraudo, Jordi; Alonso-Chamarro, Julián; Puyol, Mar

2014-10-01

While magnetic bead (MB)-based bioassays have been implemented in integrated devices, their handling on-chip is normally either not optimal--i.e. only trapping is achieved, with aggregation of the beads--or requires complex actuator systems. Herein, we describe a simple and low-cost magnetic actuator to trap and move MBs within a microfluidic chamber in order to enhance the mixing of a MB-based reaction. The magnetic actuator consists of a CD-shaped plastic unit with an arrangement of embedded magnets which, when rotating, generate the mixing. The magnetic actuator has been used to enhance the amplification reaction of an enzyme-linked fluorescence immunoassay to detect Escherichia coli O157:H7 whole cells, an enterohemorrhagic strain, which have caused several outbreaks in food and water samples. A 2.7-fold sensitivity enhancement was attained with a detection limit of 603 colony-forming units (CFU) /mL, when employing the magnetic actuator.

Electrochemical immunosensor for the determination of insulin-like growth factor-1 using electrodes modified with carbon nanotubes-poly(pyrrole propionic acid) hybrids.

Science.gov (United States)

Serafín, V; Agüí, L; Yáñez-Sedeño, P; Pingarrón, J M

2014-02-15

An amperometric immunosensor for the determination of the hormone insulin-like growth factor 1 (IGF1) is reported for the first time in this work. As electrochemical transducer, a multiwalled carbon nanotubes-modified glassy carbon electrode on which poly(pyrrole propionic acid) was electropolymerized was prepared. This approach provided a high content of surface confined carboxyl groups suitable for direct covalent binding of anti-IGF1 monoclonal antibody. A sandwich-type immunoassay using a polyclonal antibody labeled with peroxidase, hydrogen peroxide as the enzyme substrate and catechol as redox mediator was employed to monitor the affinity reaction. All the variables involved in the preparation of the modified electrode were optimized and the electrodes were characterized by electrochemical impedance spectroscopy and cyclic voltammetry. Moreover, the different experimental variables affecting the amperometric response of the immunosensor were also optimized. The calibration graph for IGF1 showed a range of linearity extending from 0.5 to 1000 pg/mL, with a detection limit, 0.25 pg/mL, more than 100 times lower than the lowest values reported for the ELISA immunoassays available for IGF1 (30 pg/mL, approximately). Excellent reproducibility for the measurements carried out with different immunosensors and selectivity against other hormones were also evidenced. A commercial human serum spiked with IGF1 at different levels between 0.01 and 10.0 ng/mL was analyzed with good results. © 2013 Elsevier B.V. All rights reserved.

History of inductively coupled plasma mass spectrometry-based immunoassays

International Nuclear Information System (INIS)

Giesen, Charlotte; Waentig, Larissa; Panne, Ulrich; Jakubowski, Norbert

2012-01-01

The analysis of biomolecules requires highly sensitive and selective detection methods capable of tolerating a complex, biological matrix. First applications of biomolecule detection by ICP-MS relied on the use of heteroelements as a label for quantification. However, the combination of immunoassays and ICP-MS facilitates multiparametric analyses through elemental tagging, and provides a powerful alternative to common bioanalytical methods. This approach extends the detection of biomarkers in clinical diagnosis, and has the potential to provide a deeper understanding of the investigated biological system. The results might lead to the detection of diseases at an early stage, or guide treatment plans. Immunoassays are well accepted and established for diagnostic purposes, albeit ICP-MS is scarcely applied for the detection of immune-based assays. However, the screening of biomarkers demands high throughput and multiplex/multiparametric techniques, considering the variety of analytes to be queried. Finally, quantitative information on the expression level of biomarkers is highly desirable to identify abnormalities in a given organism. Thus, it is the aim of this review to introduce the fundamentals, and to discuss the enormous strength of ICP-MS for the detection of different immunoassays on the basis of selected applications, with a special focus on LA‐ICP‐MS. - Highlights: ► We discuss the fundamentals of elemental tagging for ICP‐MS applications. ► We propose a definition for the expressions “label” and “tag”. ► We highlight LA‐ICP‐MS‐based heteroelement detection. ► We give an historic overview on ICP-MS and LA‐ICP‐MS-based immunoassays. ► In a personal outlook, we discuss future improvements realistically attainable.

BIOTIN INTERFERENCE WITH ROUTINE CLINICAL IMMUNOASSAYS: UNDERSTAND THE CAUSES AND MITIGATE THE RISKS.

Science.gov (United States)

Samarasinghe, Shanika; Meah, Farah; Singh, Vinita; Basit, Arshi; Emanuele, Nicholas; Emanuele, Mary Ann; Mazhari, Alaleh; Holmes, Earle W

2017-08-01

The objectives of this report are to review the mechanisms of biotin interference with streptavidin/biotin-based immunoassays, identify automated immunoassay systems vulnerable to biotin interference, describe how to estimate and minimize the risk of biotin interference in vulnerable assays, and review the literature pertaining to biotin interference in endocrine function tests. The data in the manufacturer's "Instructions for Use" for each of the methods utilized by seven immunoassay system were evaluated. We also conducted a systematic search of PubMed/MEDLINE for articles containing terms associated with biotin interference. Available original reports and case series were reviewed. Abstracts from recent scientific meetings were also identified and reviewed. The recent, marked, increase in the use of over-the-counter, high-dose biotin supplements has been accompanied by a steady increase in the number of reports of analytical interference by exogenous biotin in the immunoassays used to evaluate endocrine function. Since immunoassay methods of similar design are also used for the diagnosis and management of anemia, malignancies, autoimmune and infectious diseases, cardiac damage, etc., biotin-related analytical interference is a problem that touches every area of internal medicine. It is important for healthcare personnel to become more aware of immunoassay methods that are vulnerable to biotin interference and to consider biotin supplements as potential sources of falsely increased or decreased test results, especially in cases where a lab result does not correlate with the clinical scenario. FDA = U.S. Food & Drug Administration FT3 = free tri-iodothyronine FT4 = free thyroxine IFUs = instructions for use LH = luteinizing hormone PTH = parathyroid hormone SA/B = streptavidin/biotin TFT = thyroid function test TSH = thyroid-stimulating hormone.

Automation on an Open-Access Platform of Alzheimer's Disease Biomarker Immunoassays.

Science.gov (United States)

Gille, Benjamin; Dedeene, Lieselot; Stoops, Erik; Demeyer, Leentje; Francois, Cindy; Lefever, Stefanie; De Schaepdryver, Maxim; Brix, Britta; Vandenberghe, Rik; Tournoy, Jos; Vanderstichele, Hugo; Poesen, Koen

2018-04-01

The lack of (inter-)laboratory standardization has hampered the application of universal cutoff values for Alzheimer's disease (AD) cerebrospinal fluid (CSF) biomarkers and their transfer to general clinical practice. The automation of the AD biomarker immunoassays is suggested to generate more robust results than using manual testing. Open-access platforms will facilitate the integration of automation for novel biomarkers, allowing the introduction of the protein profiling concept. A feasibility study was performed on an automated open-access platform of the commercial immunoassays for the 42-amino-acid isoform of amyloid-β (Aβ 1-42 ), Aβ 1-40 , and total tau in CSF. Automated Aβ 1-42 , Aβ 1-40 , and tau immunoassays were performed within predefined acceptance criteria for bias and imprecision. Similar accuracy was obtained for ready-to-use calibrators as for reconstituted lyophilized kit calibrators. When compared with the addition of a standard curve in each test run, the use of a master calibrator curve, determined before and applied to each batch analysis as the standard curve, yielded an acceptable overall bias of -2.6% and -0.9% for Aβ 1-42 and Aβ 1-40 , respectively, with an imprecision profile of 6.2% and 8.4%, respectively. Our findings show that transfer of commercial manual immunoassays to fully automated open-access platforms is feasible, as it performs according to universal acceptance criteria.

Synthesis of iron oxides nanoparticles with very high saturation magnetization form TEA-Fe(III) complex via electrochemical deposition for supercapacitor applications

Science.gov (United States)

Elrouby, Mahmoud; Abdel-Mawgoud, A. M.; El-Rahman, Rehab Abd

2017-11-01

This work is devoted to the synthesis of magnetic iron oxides nanoparticles with very high saturation magnetization to be qualified for supercapacitor applications using, a simple electrodeposition technique. It is found that the electrochemical reduction process depends on concentration, temperature, deposition potential and the scan rate of potential. The nature of electrodeposition process has been characterized via voltammetric and chronoamperometric techniques. The morphology of the electrodeposits has been investigated using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The structure and phase content of these investigated electrodeposits have been examined and calculated. The obtained iron oxides show a high saturation magnetization (Ms) of about 229 emu g-1. The data exhibited a relation between Ms of electrodeposited iron oxide and specific capacitance. This relation exhibits that the highest Ms value of electrodeposited iron oxides gives also highest specific capacitance of about 725 Fg-1. Moreover, the electrodeposited iron oxides exhibit a very good stability. The new characteristics of the electro synthesized iron oxides at our optimized conditions, strongly qualify them as a valuable material for high-performance supercapacitor applications.

Recent Advances in Electrochemical Glycobiosensing

Directory of Open Access Journals (Sweden)

Germarie Sánchez-Pomales

2011-01-01

Full Text Available Biosensors based on electrochemical transduction mechanisms have recently made advances into the field of glycan analysis. These glyco-biosensors offer simple, rapid, sensitive, and economical approaches to the measurement need for rapid glycan analysis for biomarker detection, cancer and disease diagnostics, and bioprocess monitoring of therapeutic glycoproteins. Although the prevalent methods of glycan analysis (high-performance liquid chromatography, mass spectrometry, and nuclear magnetic resonance spectroscopy provide detailed identification and structural analysis of glycan species, there are significantly few low-cost, rapid glycan assays available for diagnostic and screening applications. Here we review instances in which glyco-biosensors have been used for glycan analysis using a variety of electrochemical transduction mechanisms (e.g., amperometric, potentiometric, impedimetric, and voltammetric, selective binding agents (e.g., lectins and antibodies, and redox species (e.g., enzyme substrates, inorganic, and nanomaterial.

Design and Fabrication of a PDMS Microchip Based Immunoassay

Energy Technology Data Exchange (ETDEWEB)

Shao, Guocheng; Wang, Wanjun; Wang, Jun; Lin, Yuehe

2010-07-01

In this paper, we describe the design and fabrication process of a polydimethylsiloxane (PDMS) microchip for on-chip multiplex immunoassay application. The microchip consists of a PDMS microfluidic channel layer and a micro pneumatic valve control layer. By selectively pressurizing the pneumatic microvalves, immuno reagents were controlled to flow and react in certain fluidic channel sites. Cross contamination was prevented by tightly closed valves. Our design was proposed to utilize PDMS micro channel surface as the solid phase immunoassay substrate and simultaneously detect four targets antigens on chip. Experiment result shows that 20psi valve pressure is sufficient to tightly close a 200µm wide micro channel with flow rate up to 20µl/min.

Preparation by irradiation of a solid support for enzyme immunoassay

International Nuclear Information System (INIS)

Kumakura, M.; Kaetsu, I.

1984-01-01

Reagents (immobilized anti-α-fetoprotein discs) having a porous structure were prepared for enzyme immunoassay of α-fetoprotein by radiation polymerization at low temperatures. Discs were attached to sticks for easy handling. The activity (determined by absorbance at 492 nm) of the discs varied with the hydrophilic properties and size of the discs. The discs are sufficiently sensitive and precise for enzyme immunoassay of α-fetoprotein. Anti-AFP dissolved in PBS solution was mixed with a monomer solution of hydroxyethyl methacrylate and hydroxypropyl methacrylate. The mixture was frozen to -78 0 C and gamma irradiated. (Auth.)

Novel immunoassay formats for integrated microfluidic circuits: diffusion immunoassays (DIA)

Science.gov (United States)

Weigl, Bernhard H.; Hatch, Anson; Kamholz, Andrew E.; Yager, Paul

2000-03-01

Novel designs of integrated fluidic microchips allow separations, chemical reactions, and calibration-free analytical measurements to be performed directly in very small quantities of complex samples such as whole blood and contaminated environmental samples. This technology lends itself to applications such as clinical diagnostics, including tumor marker screening, and environmental sensing in remote locations. Lab-on-a-Chip based systems offer many *advantages over traditional analytical devices: They consume extremely low volumes of both samples and reagents. Each chip is inexpensive and small. The sampling-to-result time is extremely short. They perform all analytical functions, including sampling, sample pretreatment, separation, dilution, and mixing steps, chemical reactions, and detection in an integrated microfluidic circuit. Lab-on-a-Chip systems enable the design of small, portable, rugged, low-cost, easy to use, yet extremely versatile and capable diagnostic instruments. In addition, fluids flowing in microchannels exhibit unique characteristics ('microfluidics'), which allow the design of analytical devices and assay formats that would not function on a macroscale. Existing Lab-on-a-chip technologies work very well for highly predictable and homogeneous samples common in genetic testing and drug discovery processes. One of the biggest challenges for current Labs-on-a-chip, however, is to perform analysis in the presence of the complexity and heterogeneity of actual samples such as whole blood or contaminated environmental samples. Micronics has developed a variety of Lab-on-a-Chip assays that can overcome those shortcomings. We will now present various types of novel Lab- on-a-Chip-based immunoassays, including the so-called Diffusion Immunoassays (DIA) that are based on the competitive laminar diffusion of analyte molecules and tracer molecules into a region of the chip containing antibodies that target the analyte molecules. Advantages of this

Nitrocellulose membrane-based enzyme-linked immunoassay for dengue serotype-1 IgM detection

International Nuclear Information System (INIS)

Leon, S.; Guevara, C.; Chunga, A.

1999-01-01

To evaluate the sensitivity and specifity of a nitrocellulose membrane-based immunoassay for dengue IgM, with respect to capture enzyme immunoassay, for the diagnosis of dengue virus infection. 101 serum samples were processed and divided into 2 groups: 53 from dengue serotype 1 (DEN1) infected patients, and 48 from healthy subjects. Both groups were tested with a nitrocellulose membrane-based IgM capture enzyme immunoassay (NMB-EIA) and also with an ELISA as referential pattern. NMB-EIA testing detected IgM anti-DEN1 in 94,34% of samples from infected patients, and in 14,58% of control samples, whereas ELISA fails to report false positive or false negative results: NMB-EIA appears to be a good alternative for dengue infection diagnosis. (authors)

Rapid detection of fungal alpha-amylase in the work environment with a lateral flow immunoassay

NARCIS (Netherlands)

Bogdanovic, J.; Koets, M.; Sander, I.; Wouters, I.; Meijster, T.; Heederik, D.J.J.; Amerongen, van A.; Doekes, G.

2006-01-01

Background Occupational allergen exposure assessment usually requires airborne dust sampling at the worksite followed by dust extraction and enzyme immunoassay (EIA) analysis at the laboratory. Use of semiquantitative lateral flow immunoassays (LFIAs) may allow a more rapid detection procedure with

Immunoassay separation technique

International Nuclear Information System (INIS)

1977-01-01

A method for effecting the immunoassay of a multiplicity of samples, each possibly containing an antigen or an antibody to be assayed, is discussed. Each sample is incubated with a solution containing a detectable antigen or antibody to form a multiplicity of mixtures, each mixture containing as components antigen-antibody, non-complexed antigen and non-complexed antibody. At least one of the components of the said mixture is separated by adsorption. There after, quantity of detectable antigen or antibody is detected in one of the non-adsorbed portions of the mixture. An improvement, compared to other techniques, is the continuous and sequential separation of at least one component, which is intended to be separated from each said multiplicity of mixtures

Comparison of pre-processing methods for multiplex bead-based immunoassays.

Science.gov (United States)

Rausch, Tanja K; Schillert, Arne; Ziegler, Andreas; Lüking, Angelika; Zucht, Hans-Dieter; Schulz-Knappe, Peter

2016-08-11

High throughput protein expression studies can be performed using bead-based protein immunoassays, such as the Luminex® xMAP® technology. Technical variability is inherent to these experiments and may lead to systematic bias and reduced power. To reduce technical variability, data pre-processing is performed. However, no recommendations exist for the pre-processing of Luminex® xMAP® data. We compared 37 different data pre-processing combinations of transformation and normalization methods in 42 samples on 384 analytes obtained from a multiplex immunoassay based on the Luminex® xMAP® technology. We evaluated the performance of each pre-processing approach with 6 different performance criteria. Three performance criteria were plots. All plots were evaluated by 15 independent and blinded readers. Four different combinations of transformation and normalization methods performed well as pre-processing procedure for this bead-based protein immunoassay. The following combinations of transformation and normalization were suitable for pre-processing Luminex® xMAP® data in this study: weighted Box-Cox followed by quantile or robust spline normalization (rsn), asinh transformation followed by loess normalization and Box-Cox followed by rsn.

Feasibility of a simple microsieve-based immunoassay platform

NARCIS (Netherlands)

Zweitzig, D.R.; Tibbe, Arjan G.J.; Nguyen, A.T.; van Rijn, C.J.M.; Kopnitsky, M.J.; Cichonski, K.; Terstappen, Leonardus Wendelinus Mathias Marie

2016-01-01

The intrinsic properties of silicon microsieves, such as an optically flat surface, high overall porosity, and low flow resistance have led to an increasing number of biotechnology applications. In this report, the feasibility of creating a microsieve-based immunoassay platform was explored.

Feasibility of a simple microsieve-based immunoassay platform

NARCIS (Netherlands)

Zweitzig, Daniel R.; Tibbe, Arjan G.; Nguyen, Ai T.; Rijn, van Cees J.M.; Kopnitsky, Mark J.; Cichonski, Kathleen; Terstappen, Leon W.M.M.

2016-01-01

The intrinsic properties of silicon microsieves, such as an optically flat surface, high overall porosity, and low flow resistance have led to an increasing number of biotechnology applications. In this report, the feasibility of creating a microsieve-based immunoassay platform was explored.

Alkaline phosphatase-fused repebody as a new format of immuno-reagent for an immunoassay

Energy Technology Data Exchange (ETDEWEB)

Seo, Hyo-Deok; Lee, Joong-jae [Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon, 305-701 (Korea, Republic of); Kim, Yu Jung [Industrial Biotechnology and Bioenergy Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon (Korea, Republic of); Hantschel, Oliver [School of Life Sciences, École Polytechnique Fédérale de Lausanne (EPFL), Lausanne (Switzerland); Lee, Seung-Goo [Industrial Biotechnology and Bioenergy Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon (Korea, Republic of); Kim, Hak-Sung, E-mail: hskim76@kaist.ac.kr [Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon, 305-701 (Korea, Republic of)

2017-01-15

Enzyme-linked immunoassays based on an antibody-antigen interaction are widely used in biological and medical sciences. However, the conjugation of an enzyme to antibodies needs an additional chemical process, usually resulting in randomly cross-linked molecules and a loss of the binding affinity and enzyme activity. Herein, we present the development of an alkaline phosphatase-fused repebody as a new format of immuno-reagent for immunoassays. A repebody specifically binding to human TNF-α (hTNF-α) was selected through a phage display, and its binding affinity was increased up to 49 nM using a modular engineering approach. A monomeric alkaline phosphatase (mAP), which was previously isolated from a metagenome library, was genetically fused to the repebody as a signal generator, and the resulting repebody-mAP fusion protein was used for direct and sandwich immunoassays of hTNF-α. We demonstrate the utility and potential of the repebody-mAP fusion protein as an immuno-reagent by showing the sensitivity of 216 pg mL{sup −1} for hTNF-α in a sandwich immunoassay. Furthermore, this repebody-mAP fusion protein enabled the detection of hTNF-α spiked in a serum-supplemented medium with high accuracy and reproducibility. It is thus expected that a mAP-fused repebody can be broadly used as an immuno-reagent in immunoassays. - Highlights: • A human TNF-α (hTNF-α)-specific repebody was selected using a phage display. • A monomeric alkaline phosphatase (mAP) was genetically fused to the repebody. • mAP-fused repebody enabled detection of hTNF-α with high sensitivity and accuracy. • mAP-fused repebody can be widely used as a new immuno-reagent in immunoassays.

Study on the determination of human placental lactogen (HPL) using an enzyme-immunoassay. Comparison with a commercial radio-immunoassay in the course of normal pregnancies

International Nuclear Information System (INIS)

Schneider, B.

1982-01-01

A novel enzyme-immunoassay (EIA) for determining human placental lactogen (HPL) was studied for its practicability and quality. The precision of the system in series was tested by using a serum taken each in the 19th, 29th and 40th pregnancy week. A normal range graph between the 10th and the 40th pregnancy week (10 sera per pregnancy week) was established from 310 sera of normal-course pregnancies. The graph practically agreed with the known RIA-established graphs. When comparing with a radio-immunoassay for HPL of routine application and known quality criteria, r=0.93 indicated a close correlation of the values found. (orig./MG) [de

Surface-Enhanced Raman Scattering (SERS) for Detection in Immunoassays: applications, fundamentals, and optimization

International Nuclear Information System (INIS)

Jeremy Daniel Driskell

2006-01-01

Immunoassays have been utilized for the detection of biological analytes for several decades. Many formats and detection strategies have been explored, each having unique advantages and disadvantages. More recently, surface-enhanced Raman scattering (SERS) has been introduced as a readout method for immunoassays, and has shown great potential to meet many key analytical figures of merit. This technology is in its infancy and this dissertation explores the diversity of this method as well as the mechanism responsible for surface enhancement. Approaches to reduce assay times are also investigated. Implementing the knowledge gained from these studies will lead to a more sensitive immunoassay requiring less time than its predecessors. This dissertation is organized into six sections. The first section includes a literature review of the previous work that led to this dissertation. A general overview of the different approaches to immunoassays is given, outlining the strengths and weaknesses of each. Included is a detailed review of binding kinetics, which is central for decreasing assay times. Next, the theoretical underpinnings of SERS is reviewed at its current level of understanding. Past work has argued that surface plasmon resonance (SPR) of the enhancing substrate influences the SERS signal; therefore, the SPR of the extrinsic Raman labels (ERLs) utilized in our SERS-based immunoassay is discussed. Four original research chapters follow the Introduction, each presented as separate manuscripts. Chapter 2 modifies a SERS-based immunoassay previously developed in our group, extending it to the low-level detection of viral pathogens and demonstrating its versatility in terms of analyte type, Chapter 3 investigates the influence of ERL size, material composition, and separation distance between the ERLs and capture substrate on the SERS signal. This chapter links SPR with SERS enhancement factors and is consistent with many of the results from theoretical treatments

Surface-Enhanced Raman Scattering (SERS) for Detection in Immunoassays. Applications, fundamentals, and optimization

Energy Technology Data Exchange (ETDEWEB)

Driskell, Jeremy Daniel [Iowa State Univ., Ames, IA (United States)

2006-08-09

Immunoassays have been utilized for the detection of biological analytes for several decades. Many formats and detection strategies have been explored, each having unique advantages and disadvantages. More recently, surface-enhanced Raman scattering (SERS) has been introduced as a readout method for immunoassays, and has shown great potential to meet many key analytical figures of merit. This technology is in its infancy and this dissertation explores the diversity of this method as well as the mechanism responsible for surface enhancement. Approaches to reduce assay times are also investigated. Implementing the knowledge gained from these studies will lead to a more sensitive immunoassay requiring less time than its predecessors. This dissertation is organized into six sections. The first section includes a literature review of the previous work that led to this dissertation. A general overview of the different approaches to immunoassays is given, outlining the strengths and weaknesses of each. Included is a detailed review of binding kinetics, which is central for decreasing assay times. Next, the theoretical underpinnings of SERS is reviewed at its current level of understanding. Past work has argued that surface plasmon resonance (SPR) of the enhancing substrate influences the SERS signal; therefore, the SPR of the extrinsic Raman labels (ERLs) utilized in our SERS-based immunoassay is discussed. Four original research chapters follow the Introduction, each presented as separate manuscripts. Chapter 2 modifies a SERS-based immunoassay previously developed in our group, extending it to the low-level detection of viral pathogens and demonstrating its versatility in terms of analyte type, Chapter 3 investigates the influence of ERL size, material composition, and separation distance between the ERLs and capture substrate on the SERS signal. This chapter links SPR with SERS enhancement factors and is consistent with many of the results from theoretical treatments

Current status and future developments in radiolabelled immunoassays

International Nuclear Information System (INIS)

Edwards, R.

1998-01-01

Radioisotopes are used extensively in medical practice and their use in RIA or IRMA usually represent a small proportion of the total. Radiolabelled immunoassays based on 125 I constitute a simple didactic, cost effective and robust technology which is still regarded as the reference method in many clinical applications. The IAEA has implemented many successful programmes using the ''bulk reagent'' approach, involving 68 countries in all the different regions. The main achievements have been in technology transfer with self sufficiency in production for some countries; training of large numbers of staff; quality control and quality assurance schemes; devolution of screening programmes for neonatal congenital hypothryoidism. Alternatives to the use of radioisotopic tracers are constrained by many factors and are often only available in restricted commercial packages. They are often not suitable for technology transfer programmes and often lack any didactic component in addition to a relative high cost. The production of radiolabels using 125 I is both simple and adaptable. In addition expertise in their preparation and purification is widespread even in developing countries. Together with the ease of producing antibodies, the facts have made 125 I-radiolabelled immunoassays ideal for investigative procedures for many research activities (30,31) particularly in the medical context where radioisotopes are commonly used. In conclusion, even a superficial examination of public health statistics for various countries throughout the continents indicates a need for a simple, inexpensive and robust analytical tool. In this light, there is a predicted continuing role for radiolabelled immunoassays. (author)

Evaluation of six immunoassays for detection of dengue virus-specific immunoglobulin M and G antibodies

NARCIS (Netherlands)

J. Groen (Jan); P. Koraka (Penelope); J. Velzing (Jans); C. Copra (Cederick); A.D.M.E. Osterhaus (Albert)

2000-01-01

textabstractThe performance of six commercially available immunoassay systems for the detection of dengue virus-specific immunoglobulin M (IgM) and IgG antibodies in serum was evaluated. These included two IgM and IgG enzyme immunoassays (EIA) from MRL Laboratories and PanBio, a rapid

An Inexpensive, Fast and Sensitive Quantitative Lateral Flow Magneto-Immunoassay for Total Prostate Specific Antigen

Directory of Open Access Journals (Sweden)

Jacqueline M. Barnett

2014-07-01

Full Text Available We describe the detection characteristics of a device the Resonant Coil Magnetometer (RCM to quantify paramagnetic particles (PMPs in immunochromatographic (lateral flow assays. Lateral flow assays were developed using PMPs for the measurement of total prostate specific antigen (PSA in serum samples. A detection limit of 0.8 ng/mL was achieved for total PSA using the RCM and is at clinically significant concentrations. Comparison of data obtained in a pilot study from the analysis of serum samples with commercially available immunoassays shows good agreement. The development of a quantitative magneto-immunoassay in lateral flow format for total PSA suggests the potential of the RCM to operate with many immunoassay formats. The RCM has the potential to be modified to quantify multiple analytes in this format. This research shows promise for the development of an inexpensive device capable of quantifying multiple analytes at the point-of-care using a magneto-immunoassay in lateral flow format.

Type I Clathrates as Novel Silicon Anodes: An Electrochemical and Structural Investigation

OpenAIRE

Li, Ying; Raghavan, Rahul; Wagner, Nicholas A.; Davidowski, Stephen K.; Baggetto, Lo?c; Zhao, Ran; Cheng, Qian; Yarger, Jeffery L.; Veith, Gabriel M.; Ellis?Terrell, Carol; Miller, Michael A.; Chan, Kwai S.; Chan, Candace K.

2015-01-01

Silicon clathrates contain cage?like structures that can encapsulate various guest atoms or molecules. An electrochemical evaluation of type I silicon clathrates based on Ba8Al y Si46?y as the anode material for lithium?ion batteries is presented here. Postcycling characterization with nuclear magnetic resonance and X?ray diffraction shows no discernible structural or volume changes even after electrochemical insertion of 44 Li (?1 Li/Si) into the clathrate structure. The observed properties ...

Fundamentals of magnet-actuated droplet manipulation on an open hydrophobic surface†

Science.gov (United States)

Long, Zhicheng; Shetty, Abhishek M.; Solomon, Michael J.; Larson, Ronald G.

2010-01-01

We systematically investigate droplet movement, coalescence, and splitting on an open hydrophobic surface. These processes are actuated by magnetic beads internalized in an oil-coated aqueous droplet using an external magnet. Results are organized into an ‘operating diagram’ that describes regions of droplet stable motion, breakage, and release from the magnet. The results are explained theoretically with a simple model that balances magnetic, friction, and capillary-induced drag forces and includes the effects of particle type, droplet size, surrounding oil layer, surface tension, and viscosity. Finally, we discuss the implications of the results for the design of magnet-actuated droplet systems for applications such as nucleic acid purification, immunoassay and drug delivery. PMID:19458864

A fast universal immobilization of immunoglobulin G at 4 °C for the development of array-based immunoassays.

Directory of Open Access Journals (Sweden)

Shu-Lin Guo

Full Text Available To maintain the antibody activity and enhance performance of array-based immunoassays, protein G was used to allow a shorter duration of immunoglobulin G immobilization at 4 °C, with the antibody placed in the appropriate orientation. The multiplexed detection of six pain-related message molecules (PRMMs was used as examples for the development of array-based immunoassays: substance P, calcitonin gene-related peptide, nerve growth factor, brain-derived neurotrophic factor, tumor necrosis factor-α, and β-endorphin. Protein G- and non-protein G-coated slides were tested. Compared to non-protein G immunoassays, protein G shortened the antibody immobilization time at 4 °C from overnight to 2 hours. Only protein G-facilitated immunoassays succeeded in simultaneously detecting all six PRMMs with high specificity. Dose-response curves showed that the limits of detection of the protein G-multiplexed immunoassays for the PRMMs was approximately 164, 167, 120, 60, 80, and 92 pg/ml, respectively. Thus, protein G effectively shortens the duration of antibody immobilization at 4 °C, allowing the use of sensitive array-based immunoassays for the simultaneous detection of PRMMs.

High temperature radio-frequency superconducting quantum interference device system for detection of magnetic nanoparticles

International Nuclear Information System (INIS)

Pretzell, Alf

2012-01-01

This doctoral thesis was aimed at establishing a set-up with high-temperature superconductor (HTS) radio-frequency (rf) superconducting quantum interference device (SQUID) technology for the detection of magnetic nanoparticles and in particular for testing applications of magnetic nanoparticle immunoassays. It was part of the EU-project ''Biodiagnostics'' running from 2005 to 2008. The method of magnetic binding assays was developed as an alternative to other methods of concentration determination like enzyme linked immunosorbent assay (ELISA), or fluorescent immunoassay. The ELISA has sensitivities down to analyte-concentrations of pg/ml. Multiple incubation and washing steps have to be performed for these techniques, the analyte has to diffuse to the site of binding. The magnetic assay uses magnetic nanoparticles as markers for the substance to be detected. It is being explored by current research and shows similar sensitivity compared to ELISA but in contrast - does not need any washing and can be read out directly after binding - can be applied in solution with opaque media, e.g. blood or muddy water - additionally allows magnetic separation or concentration - in combination with small magnetoresistive or Hall sensors, allows detection of only a few particles or even single beads. For medical or environmental samples, maybe opaque and containing a multitude of substances, it would be advantageous to devise an instrument, which allows to be read out quickly and with high sensitivity. Due to the mentioned items the magnetic assay might be a possibility here.

Comparative determination of phenytoin by spectrophotometry, gas chromatography, liquid chromatography, enzyme immunoassay, and radioimmunoassay

International Nuclear Information System (INIS)

Castro, A.; Ibanez, J.; DiCesare, J.L.; Adams, R.F.; Malkus, H.

1978-01-01

Sera from patients being treated with phenytoin were analyzed for the drug by spectrophotometry, gas chromatography, radioimmunoasay, enzyme immunoassay, and liquid chromatography. The assay values obtained were intercompared statistically. Enzyme immunoassay and liquid chromatography appear to be attractive alternatives to the more traditional methods of spectrophotometry and gas chromatography. Our radioimmunoassay data correlated poorly with results by the four other methods

Fabrication and electrochemical performance of graphene—ZnO nanocomposites

International Nuclear Information System (INIS)

Li Zhen-Peng; Men Chuan-Ling; Wang Wan; Cao Jun

2014-01-01

Graphene—ZnO nanocomposites were synthesized successfully through a one-step solvothermal approach. The morphology, structure, and composition of the prepared nanocomposites were investigated by scanning electron microscopy (SEM), transmission electron microscope (TEM), laser micro Raman spectroscopy, and Fourier transform infra-red spectroscopy (FT-IR). The outcomes confirmed that this approach is comparatively steady, practicable, and operable compared with other reported methods. The electrochemical performance of the graphene-ZnO electrodes was analyzed through cyclic voltammetry, altering-current (AC) impedance, and chronopotentiometry tests. The graphene—ZnO electrodes exhibited an improved electrode performance with higher specific capacitance (115 F·g −1 ), higher electrochemical stability, and higher energy density than the graphene electrodes and most reported graphene—ZnO electrodes. Graphene—ZnO nanocomposites have a steady reversible charge/discharge behavior, which makes them promising candidates for electrochemical capacitors (ECs). (condensed matter: electronic structure, electrical, magnetic, and optical properties)

Which amphetamine-type stimulants can be detected by oral fluid immunoassays?

Science.gov (United States)

Souza, Daniele Z; Boehl, Paula O; Comiran, Eloisa; Prusch, Débora S; Zancanaro, Ivomar; Fuentefria, Alexandre M; Pechansky, Flavio; Duarte, Paulina C A V; De Boni, Raquel B; Fröehlich, Pedro E; Limberger, Renata P

2012-02-01

The use of oral fluid for monitoring drug consumption on roads has many advantages over conventional biological fluids; therefore, several immunoassays have been developed for this purpose. In this work, the ability of 3 commercial immunoassays to detect amphetamine-type stimulants (ATSs) in oral fluid was assessed. In addition, it was reviewed the main controlled ATSs available worldwide, as well as the oral fluid immunological screening tests that have been used for identifying ATSs in drivers. The analytical specificity of amphetamine direct enzyme-linked immunosorbent assay (ELISA), methamphetamine direct ELISA (Immunalysis Corporation), and Oral-View saliva multidrug of abuse test (Alfa Scientific Designs) was evaluated using ATS-spiked oral fluid. Legislation and published articles that report the use of immunological screening tests to detect ATS consumption in conductors were reviewed, including the kit's technical information, project reports, police and drug databases. Even at high concentrations, the tested assays were not able to detect methylphenidate, fenproporex, or diethylpropion, controlled ATSs legally marketed in many countries. This evidences the need to develop new kits that enable one to control the misuse of prescription ATSs on roads through oral fluid immunoassays.

Plasmon enhanced fluoro-immunoassay using egg yolk antibodies for ultra-sensitive detection of herbicide diuron.

Science.gov (United States)

Sharma, Priyanka; Kukkar, Manil; Ganguli, Ashok K; Bhasin, Aman; Suri, C Raman

2013-08-07

Plasmon enhanced fluorescence immunoassay (PEFI) format has been reported in developing a sensitive heterogeneous fluoroimmunoassay for monitoring the phenylurea herbicide diuron. Computer-assisted molecular modeling was carried out to study the conformational and electrostatic effects of synthesized hapten for producing highly specific egg yolk antibody against a phenyl urea herbicide diuron. The generated antibodies were labeled with fluorescein isothiocyanate at different molar ratios and used as tracer in the developed fluorescence based immunoassay. The sensitivity of the assay format was enhanced by using silver nanoparticles tagged with bovine serum albumin as a new blocking reagent in the developed PEFI format. Enhancer treatment on the developed immunoassay showed a significant improvement of fluorescence signal intensity with approximately 10 fold increase in assay sensitivity. The immunoassay has a detection limit of 0.01 ng mL(-1) with good signal precision (~2%) in the optimum working concentration range between 1 pg mL(-1) to 10 μg mL(-1) of diuron. These findings facilitate high throughput fluorescence-based processes that could be useful in biology, drug discovery and compound screening applications.

Fake news? Biotin interference in thyroid immunoassays.

Science.gov (United States)

Koehler, Viktoria F; Mann, Ulrike; Nassour, Ayham; Alexander Mann, W

2018-05-29

We report on a 47 year old male patient with multiple sclerosis (MS) presenting in our outpatient neurology clinic in Frankfurt/Main for therapy evaluation. Before change of treatment laboratory investigations were performed. Thyroid function tests (TFTs) with a streptavidin/biotin based immunoassay revealed severe hyperthyroidism with positive thyroid autoantibodies suggestive for Graves' disease. Clinical presentation and thyroid sonography were unremarkable. Due to the discordance between clinical presentation and TFTs, we repeated medical history, in which the patient reported taking high-doses of biotin (300 mg/day) for MS. Recent studies with patients suffering from primary and secondary progressive MS, indicated promising effects of high-dose biotin on MS-related disability. In immunoassays relaying on streptavidin-biotin interaction, biotin intake can cause falsely high or low results. Two weeks after withdrawing biotin, biotin/streptavidin dependant assays showed no longer the biochemical picture of severe hyperthyroidism. Biotin intake should be paused for at least two to five days prior to the use of biotin/streptavidin dependant assays. Alternatively, non-biotin/streptavidin dependant assays (radioimmunoassay, gas chromatography-mass spectrometry/liquid chromatography-mass spectrometry) may be used. Copyright © 2017. Published by Elsevier B.V.

An environmentally friendly electro-oxidative approach to recover valuable elements from NdFeB magnet waste

NARCIS (Netherlands)

Venkatesan, P.; Sun, Z.; Sietsma, J.; Yang, Y.

2018-01-01

In this manuscript, we demonstrate a room temperature electrochemical process for efficiently recycling NdFeB magnet waste. First, the magnet waste was completely leached with HCl and then, in-situ electrochemical oxidation was performed to selectively oxidize Fe(II) in the leachate to Fe(III).

Preparation of Pr-Fe-Co-B-Nb-M (M= Al, P, Cu, Ga and/or Gd) HDDR magnets and alloys and characterization of their magnetic properties and corrosion resistance

International Nuclear Information System (INIS)

Oliveira, Mara Cristina Lopes de

2009-01-01

HDDR process has attracted great interest for producing polymer- bonded rare earth based magnets. It presents commercial advantages when compared with conventional sintered magnets owing to easy and low cost manufacturing. With the development of anisotropic powders using praseodymium, the expectations about this process grow e also the need for studying new compositions and alloy additions. In this work the magnetic properties of polymer-bonded magnets prepared with PrFeB magnetic alloys using HDDR process have been studied. Pr 14 Fe bal Co 16 B 6 Nb 0,1 was used as the reference alloy Phosphorus, copper, aluminium, gallium and gadolinium additions have been performed to increase the magnetic properties of the reference alloy. The microstructural characterization of the magnets has been carried out through optical microscopy and SEM. The complex microstructure influences the electrochemical behavior of the magnetic alloys. The literature about this subject is scarce. Thus, the corrosion resistance of the different alloys prepared during this work was evaluated using electrochemical impedance spectroscopy and potentiodynamic polarization curves. A correlation between the microstructural features and the electrochemical behavior of the alloys has been established. The results showed that phosphorus and aluminium additions up to 1.0wt% had a beneficial effect on the magnetic properties and corrosion resistance of the alloys. Copper additions, on the other hand, strongly diminished the magnetic properties of the reference alloy. (author)

Electrochemical Biosensors - Sensor Principles and Architectures

Science.gov (United States)

Grieshaber, Dorothee; MacKenzie, Robert; Vörös, Janos; Reimhult, Erik

2008-01-01

Quantification of biological or biochemical processes are of utmost importance for medical, biological and biotechnological applications. However, converting the biological information to an easily processed electronic signal is challenging due to the complexity of connecting an electronic device directly to a biological environment. Electrochemical biosensors provide an attractive means to analyze the content of a biological sample due to the direct conversion of a biological event to an electronic signal. Over the past decades several sensing concepts and related devices have been developed. In this review, the most common traditional techniques, such as cyclic voltammetry, chronoamperometry, chronopotentiometry, impedance spectroscopy, and various field-effect transistor based methods are presented along with selected promising novel approaches, such as nanowire or magnetic nanoparticle-based biosensing. Additional measurement techniques, which have been shown useful in combination with electrochemical detection, are also summarized, such as the electrochemical versions of surface plasmon resonance, optical waveguide lightmode spectroscopy, ellipsometry, quartz crystal microbalance, and scanning probe microscopy. The signal transduction and the general performance of electrochemical sensors are often determined by the surface architectures that connect the sensing element to the biological sample at the nanometer scale. The most common surface modification techniques, the various electrochemical transduction mechanisms, and the choice of the recognition receptor molecules all influence the ultimate sensitivity of the sensor. New nanotechnology-based approaches, such as the use of engineered ion-channels in lipid bilayers, the encapsulation of enzymes into vesicles, polymersomes, or polyelectrolyte capsules provide additional possibilities for signal amplification. In particular, this review highlights the importance of the precise control over the delicate

A conventional chemical reaction for use in an unconventional assay: A colorimetric immunoassay for aflatoxin B1 by using enzyme-responsive just-in-time generation of a MnO2 based nanocatalyst.

Science.gov (United States)

Lai, Wenqiang; Zeng, Qiao; Tang, Juan; Zhang, Maosheng; Tang, Dianping

2018-01-10

The authors describe a colorimetric immunoassay for the model nalyte aflatoxin B 1 (AFB 1 ). It is based on the just-in-time generation of an MnO 2 nanocatalyst. Unlike previously developed immunoassay, the chromogenic reaction relies on the just-in-time formation of an oxidase mimic without the aid of the substrate. Potassium permanganate (KMnO 4 ) is converted into manganese dioxide (MnO 2 ) which acts as an oxidase mimic that catalyzes the oxidation 3,3',5,5'-tetramethylbenzidine (TMB) by oxygen to give a blue colored product. In the presence of ascorbic acid (AA), KMnO 4 is reduced to Mn(II) ions. This results in a decrease in the amount of MnO 2 nanocatalyst. Hence, the oxidation of TMB does not take place. By adding ascorbate oxidase, AA is converted into dehydroascorbic acid which cannot reduce KMnO 4 . Based on these observations, a colorimetric competitive enzyme immunoassay was developed where ascorbate oxidase and gold nanoparticle-labeled antibody against AFB 1 and magnetic beads carrying bovine serum albumin conjugated to AFB 1 are used for the determination of AFB 1 . In presence of AFB 1 , it will compete with the BSA-conjugated AFB 1 (on the magnetic beads) for the labeled antibody against AFB 1 on the gold nanoparticles. This makes the amount of ascorbate oxidase/anti-AFB 1 antibody-labeled gold nanoparticles, which conjugated on magnetic beads, reduce, and resulted in an increase of ascorbic acid. Under optimal conditions, the absorbance (measured at 652 nm) decreases with increasing AFB 1 concentrations in the range from 0.1 to 100 ng mL -1 , with a 0.1 ng mL -1 detection limit (at the 3S blank level). The accuracy of the assay was validated by analyzing spiked peanut samples. The results matched well with those obtained with a commercial ELISA kit. Conceivably, the method is not limited to aflatoxins but has a wide scope in that it may be applied to many other analytes for which respective antibodies are available. Graphical abstract

An Ultrasensitive Electrochemiluminescence Immunoassay for Carbohydrate Antigen 19-9 in Serum Based on Antibody Labeled Fe3O4 Nanoparticles as Capture Probes and Graphene/CdTe Quantum Dot Bionanoconjugates as Signal Amplifiers

Science.gov (United States)

Gan, Ning; Zhou, Jing; Xiong, Ping; Li, Tianhua; Jiang, Shan; Cao, Yuting; Jiang, Qianli

2013-01-01

The CdTe quantum dots (QDs), graphene nanocomposite (CdTe-G) and dextran–Fe3O4 magnetic nanoparticles have been synthesized for developing an ultrasensitive electrochemiluminescence (ECL) immunoassay for Carcinoembryonic antigen 19-9 (CA 19-9) in serums. Firstly, the capture probes (CA 19-9 Ab1/Fe3O4) for enriching CA 19-9 were synthesized by immobilizing the CA 19-9’s first antibody (CA 19-9 Ab1) on magnetic nanoparticles (dextran-Fe3O4). Secondly, the signal probes (CA 19-9 Ab2/CdTe-G), which can emit an ECL signal, were formed by attaching the secondary CA 19-9 antibody (CA 19-9 Ab2) to the surface of the CdTe-G. Thirdly, the above two probes were used for conjugating with a serial of CA 19-9 concentrations. Graphene can immobilize dozens of CdTe QDs on their surface, which can emit stronger ECL intensity than CdTe QDs. Based on the amplified signal, ultrasensitive antigen detection can be realized. Under the optimal conditions, the ECL signal depended linearly on the logarithm of CA 19-9 concentration from 0.005 to 100 pg/mL, and the detection limit was 0.002 pg/mL. Finally, five samples of human serum were tested, and the results were compared with a time-resolved fluorescence assay (TRFA). The novel immunoassay provides a stable, specific and highly sensitive immunoassay protocol for tumor marker detection at very low levels, which can be applied in early diagnosis of tumor. PMID:23685872

The management of isolated positive syphilis enzyme immunoassay results in HIV-negative patients attending a sexual health clinic.

Science.gov (United States)

Thorley, Nicola; Adebayo, Michael; Smit, Erasmus; Radcliffe, Keith

2016-08-01

An unconfirmed positive treponemal enzyme immunoassay (enzyme immunoassay positive, Treponema pallidum particle agglutination negative and rapid plasma reagin negative) presents a clinical challenge to distinguish early syphilis infection from false-positive results. These cases are referred for syphilis line assay (INNO-LIA) and recalled for repeat syphilis serology. We performed a retrospective audit to establish the proportion of HIV-negative cases with unconfirmed positive enzyme immunoassay results, the proportion of these cases that received an INNO-LIA test and repeat syphilis serology testing and reviewed the clinical outcomes; 0.35% (80/22687) cases had an unconfirmed positive treponemal enzyme immunoassay result. Repeat syphilis serology was performed in 80% (64/80) cases, but no additional cases of syphilis were identified. Eighty-eight per cent (70/80) received an INNO-LIA test; 14% (5/37) unconfirmed enzyme immunoassay-positive cases with no prior history of syphilis were confirmed on INNO-LIA assay, supporting a diagnosis of latent syphilis. As a confirmatory treponemal test, the INNO-LIA assay may be more useful than repeat syphilis serological testing. © The Author(s) 2016.

Simple patterned nanofiber scaffolds and its enhanced performance in immunoassay.

Directory of Open Access Journals (Sweden)

Jing Wang

Full Text Available Cancer has become the leading cause of death worldwide; early diagnosis and treatment of cancers is critical for the survival of the patients. The concentration of cancer markers in easy-to-access biological fluids can provide great assistance in screening for occult primary cancers, distinguishing malignant from benign findings, determining prognosis and prediction for cancer patients. The multiplex detection technology of a panel of cancer markers can greatly increase the accuracy of disease diagnosis. Herein, we briefly fabricate a high-throughput micro-immunoassay based on the electrospun polystyrene (PS substrates to improve detection sensitivity. The immunoassay was evaluated by analyzing three different cancer biomarkers (AFP, CEA, VEGF. For AFP, CEA, VEGF immunofluorescence assay, the LOD of assay conducted on electrospun PS substrates before or after plasma and the conventional PS substrates were 0.42, 0.10, 1.12 ng/mL, 0.57, 0.09, 1.24 ng/mL, and 159.75, 26.19, 385.59 pg/mL, respectively (P < 0.05. Due to the high porosity and large surface area-to-volume ratio which is the foremost merit of nanostructures, and the plasma treatment which make the hydrophobic PS nanofibers hydropholic, the nanofibers substrates showed sufficient retention of immunoassay functionality and high potential for capture molecules immobilization. Consequently, the immunofluorescence assay conducted on electrospun PS substrates could significantly enhance the sensitivity and limits of detection.

Deoxynivalenol-mimic nanobody isolated from a naïve phage display nanobody library and its application in immunoassay.

Science.gov (United States)

Qiu, Yu-Lou; He, Qing-Hua; Xu, Yang; Bhunia, Arun K; Tu, Zhui; Chen, Bo; Liu, Yuan-Yuan

2015-08-05

In this study, using mycotoxin deoxynivalenol (DON) as a model hapten, we developed a nanobody-based environmental friendly immunoassay for sensitive detection of DON. Two nanobodies (N-28 and N-31) which bind to anti-DON monoclonal antibody (MAb) were isolated from a naive phage display library. These nanobodies are clonable, thermally stable and mycotoxin-free products and can be served as coating antigen mimetics in heterologous immunoassay. The half inhibition concentration (IC50) of the immunoassay developed with N-28 and N-31 was 8.77 ± 0.41 ng mL(-1) and 19.97 ± 0.84 ng mL(-1), respectively, which were 18- and 8-fold more sensitive than the conventional coating antigen (DON-BSA) based immunoassay. In order to better understand the molecular mechanism of antigen mimicry by nanobody, the 3D structure of "nanobody (N-28) - anti-DON MAb" complex was presented and verified by molecular modeling and alanine-scanning mutagenesis. The results showed that hydrogen bond and hydrophobic interaction formed between Thr 102 - Ser 106 of N-28 and CDR H3 residues of anti-DON antibody may contribute to their binding. This novel concept of enhancing sensitivity of immunoassay for DON based on nanobody may provide potential applications in a general method for immunoassay of various food chemical contaminants. Copyright © 2015 Elsevier B.V. All rights reserved.

A Reuse Evaluation for Solar-Cell Silicon Wafers via Shift Revolution and Tool Rotation Using Magnetic Assistance in Ultrasonic Electrochemical Micromachining

Directory of Open Access Journals (Sweden)

P. S. Pa

2013-01-01

Full Text Available A new reuse fabrication using a tool module with rotation and revolution through a process of magnetic assistance in ultrasonic electrochemical micromachining (UEMM for removal of the surface layers from silicon wafers of solar cells is demonstrated. The target of the proposed reuse fabrication method is to replace the current approach, which uses strong acid and grinding and may damage the physical structure of silicon wafers and pollute to the environment. A precisely engineered clean production approach to removal of surface microstructure layers from silicon wafers is to develop a mass production system for recycling defective or discarded silicon wafers of solar cells that can reduce pollution and cost. The high revolution speed of the shift with the high rotation speed of the designed tool increases the discharge mobility and improves the removal effect associated with the high feed rate of the workpiece. High frequency and high power of ultrasonic with large electrolyte flow rate and high magnetic strengths with a small distance between the two magnets provide a large discharge effect and good removal; only a short period of time is required to remove the epoxy film and Si3N4 layer easily and cleanly.

Optimized Lateral Flow Immunoassay Reader for the Detection of Infectious Diseases in Developing Countries.

Science.gov (United States)

Pilavaki, Evdokia; Demosthenous, Andreas

2017-11-20

Detection and control of infectious diseases is a major problem, especially in developing countries. Lateral flow immunoassays can be used with great success for the detection of infectious diseases. However, for the quantification of their results an electronic reader is required. This paper presents an optimized handheld electronic reader for developing countries. It features a potentially low-cost, low-power, battery-operated device with no added optical accessories. The operation of this proof of concept device is based on measuring the reflected light from the lateral flow immunoassay and translating it into the concentration of the specific analyte of interest. Characterization of the surface of the lateral flow immunoassay has been performed in order to accurately model its response to the incident light. Ray trace simulations have been performed to optimize the system and achieve maximum sensitivity by placing all the components in optimum positions. A microcontroller enables all the signal processing to be performed on the device and a Bluetooth module allows transmission of the results wirelessly to a mobile phone app. Its performance has been validated using lateral flow immunoassays with influenza A nucleoprotein in the concentration range of 0.5 ng/mL to 200 ng/mL.

Optimized Lateral Flow Immunoassay Reader for the Detection of Infectious Diseases in Developing Countries

Directory of Open Access Journals (Sweden)

Evdokia Pilavaki

2017-11-01

Full Text Available Detection and control of infectious diseases is a major problem, especially in developing countries. Lateral flow immunoassays can be used with great success for the detection of infectious diseases. However, for the quantification of their results an electronic reader is required. This paper presents an optimized handheld electronic reader for developing countries. It features a potentially low-cost, low-power, battery-operated device with no added optical accessories. The operation of this proof of concept device is based on measuring the reflected light from the lateral flow immunoassay and translating it into the concentration of the specific analyte of interest. Characterization of the surface of the lateral flow immunoassay has been performed in order to accurately model its response to the incident light. Ray trace simulations have been performed to optimize the system and achieve maximum sensitivity by placing all the components in optimum positions. A microcontroller enables all the signal processing to be performed on the device and a Bluetooth module allows transmission of the results wirelessly to a mobile phone app. Its performance has been validated using lateral flow immunoassays with influenza A nucleoprotein in the concentration range of 0.5 ng/mL to 200 ng/mL.

IMMUNOASSAYS FOR THE DETECTION OF UROKINASE RECEPTOR FORMS

DEFF Research Database (Denmark)

2004-01-01

amounts of uPAR forms in a sample with data indicative of the presence of the cancer disease. The method can be used for staging, prognosis or diagnosis of prostate cancer. Two novel monoclonal antibodies and a kit and immunoassays for detecting at least one uPAR form(s) are also described in the present...

Direct salivary cortisol radio-immunoassay determination. Clinical applications

International Nuclear Information System (INIS)

Simon, C.; Cherfan, J.; Kurtz, F.; Vignon, F.; Schlienger, J.L.; Chabrier, G.

1987-01-01

Salivary cortisol levels reflect the biologically active free fraction of blood cortisol. The authors describe the results obtained with the aim of a radio-immunoassay commercial serum cortisol kit, without prealable extraction in different physiological and pathological situations. Salivary cortisol determination appears performant both in nycthemeral studies and in stimulation or freination tests [fr

Sustainable nanocomposites toward electrochemical energy storage and environmental remediation

Science.gov (United States)

Zhu, Jiahua

Energy shortage and environmental pollution are the two most concerns right now for the long term sustainable development of human society. New technology developments are the key solutions to these challenges, which strongly rely on the continuous upgrading of advanced material performance. In this dissertation, sustainable nanocomposites with multifunctionalities are designed and fabricated targeting to the applications in high energy/power density capacitor electrodes and efficient heavy metal adsorbent for polluted water purification. Contrary to the helical carbon structure from pure cotton fabrics under microwave heating and radical oxidized ignition of nanoparticles from conventional heating, magnetic carbon tubular nanocomposite fabrics decorated with unifromally dispersed Co-Co3O4 nanoparticles were successfully synthesized via a microwave heating process using cotton fabric and inorganic salt as precursors, which have shown better anti-corrosive performance and demonstrated great potential as novel electrochemical pseudocapacitor electrode. Polyaniline nanofibers (PANI-NFs)/graphite oxide (GO) nanocomposites with excellent interfacial interaction and elongated fiber structure were synthesized via a facile interfacial polymerization method. The PANI-NFs/GO hybrid materials showed orders of magnitude enhancement in capacitance and energy density than that of individual GO and PANI-NF components. At the same weight loading of PANI in the composites, fibrous PANI demonstrated higher energy density and long term stability than that of particle-shaped PANI at higher power density. Besides the efforts focusing on the inside of the capacitor including new electrodes, electrolyte materials, and capacitor configuration designs. A significant small external magnetic field (720 Gauss) induced capacitance enhancement is reported for graphene and graphene nanocomposite electrodes. The capacitance of Fe2O3/graphene nanocomposites increases by 154.6% after appling

Microfluidic "Pouch" Chips for Immunoassays and Nucleic Acid Amplification Tests.

Science.gov (United States)

Mauk, Michael G; Liu, Changchun; Qiu, Xianbo; Chen, Dafeng; Song, Jinzhao; Bau, Haim H

2017-01-01

Microfluidic cassettes ("chips") for processing and analysis of clinical specimens and other sample types facilitate point-of-care (POC) immunoassays and nucleic acid based amplification tests. These single-use test chips can be self-contained and made amenable to autonomous operation-reducing or eliminating supporting instrumentation-by incorporating laminated, pliable "pouch" and membrane structures for fluid storage, pumping, mixing, and flow control. Materials and methods for integrating flexible pouch compartments and diaphragm valves into hard plastic (e.g., acrylic and polycarbonate) microfluidic "chips" for reagent storage, fluid actuation, and flow control are described. We review several versions of these pouch chips for immunoassay and nucleic acid amplification tests, and describe related fabrication techniques. These protocols thus offer a "toolbox" of methods for storage, pumping, and flow control functions in microfluidic devices.

A sandwich immunoassay for human prolyl 4-hydroxylase using monoclonal antibody

International Nuclear Information System (INIS)

Yoshida, Shinichi

1986-01-01

Monoclonal antibody was used in a sandwich enzyme immunoassay and in a radioimmunoassay for human serum immunoreactive prolyl 4-hydroxylase. The enzyme immunoassay utilized a monoclonal antibody as a solid phase and horseradish peroxidase-labeled rabbit antibody to human prolyl 4-hydroxylase as a conjugate. Sensitivity was 0.1 ng of enzyme per tube. With a conjugate purified by an enzyme-bound affinity column, sensitivity was increased to 0.01 ng per tube, and linearity was obtained between 0.01 to 30 ng per tube. The radioimmunoassay used a 125 I-labeled rabbit antibody (IgG) as the conjugate. Sensitivity of this technique was 0.4 ng of enzyme per tube. (Auth.)

Immunoassay of β-endorphin

International Nuclear Information System (INIS)

Hoellt, V.; Gramsch, C.; Herz, A.

1979-01-01

The present paper describes the characteristics of a series of antisera against β-endorphin (β-E) developed in our laboratory which all recognize β-lipotropin and their use in (1) the determination of β-E in tissue and in body fluids and in (2) the immunocytochemical localization of β-E containing neurons in the rat brain and in (3) the study of the conversion of the β-E/β-LPH precursor into β-LPH and β-E in the pars intermedia/nervosa of the rat pituitary. It is the purpose of the present report to critically analyze the pitfalls and drawbacks, as well as the advantage, of the use of radioimmunoassay and other immunoassays in the determination of β-E. (Auth.)

Towards the development of a radioenzyme-immunoassay

International Nuclear Information System (INIS)

Schuurs, A.H.W.M.; Waart, M. v. d.

1976-01-01

We have tried to develop a very sensitive enzyme-immunoassay. For this purpose, a very sensitive radiochemical enzyme assay was used. HCG was chosen as test model and AChE as labelling enzyme. The test appeared to be much more sensitive than the normal enzymeimmunoassay. And, in comparison with RIA, it was about as sensitive but less time-consuming, and it makes use, in principle, of stable reagents. (orig./GSE) [de

Performance of hepatitis B assays on the Bayer ADVIA Centaur Immunoassay System.

Science.gov (United States)

van Helden, Josef; Denoyel, Gérard; Karwowska, Sylwia; Reamer, Randy; Schmalz, John; Wright, Ted; Preisel-Simmons, Barbara

2004-01-01

Bayer HealthCare LLC, Diagnostics Division, has developed several new assays on the ADVIA Centaur immunoassay system for the detection of markers of hepatitis B virus infection in human serum and plasma. This panel includes assays for: hepatitis B surface antigen (HBsAg), a confirmatory test method for HBsAg, antibodies to hepatitis B surface antigen (anti-HBs), IgM and IgG antibodies to hepatitis B core antigen (anti-HBc Total) and IgM antibodies to hepatitis B core antigen (anti-HBc IgM). These assays employ magnetic particle separation technology with direct chemiluminescence for optimal assay performance. All of the assays are fully automated, require sample volumes ranging from 15 microl to 100 microl (with the exception of the ADVIA Centaur HBsAg Confirmatory Assay, which requires 2 x 100 microl), and have throughputs of up to 240 tests per hour. The five ADVIA Centaur HBV assays were tested in extensive performance evaluations conducted at two sites in Europe. The performance evaluations, which included samples from HBV-infected individuals, blood donors, hospitalized/clinical patients, and HBV vaccinees (for Anti-HBs evaluation), generated performance data in support of obtaining the Communautés Européennes (CE) mark for European market distribution. The HBV performance evaluations resulted in an overall diagnostic specificity > 99%, i.e. 99.94% for the ADVIA Centaur HBsAg Assay, 100% for the ADVIA Centaur Anti-HBs Assay, 100% for the ADVIA Centaur HBc IgM Assay and 99.94% for the ADVIA Centaur HBc Total Assay. All of the ADVIA Centaur assays showed a very good diagnostic sensitivity on these populations with 100% for the ADVIA Centaur HBsAg Assay, 99.0% for the ADVIA Centaur Anti-HBs Assay, 98.53% for the ADVIA Centaur HBc IgM Assay and 100% for the ADVIA Centaur HBc Total Assay. The ADVIA Centaur HBsAg Confirmatory Test confirmed 100% of the positive HBsAg samples. Testing of interfering substances and potential cross-reacting samples for all ADVIA

Modification of a deoxynivalenol-antigen-mimicking nanobody to improve immunoassay sensitivity by site-saturation mutagenesis.

Science.gov (United States)

Qiu, Yu-Lou; He, Qing-Hua; Xu, Yang; Wang, Wei; Liu, Yuan-Yuan

2016-01-01

A nanobody (N-28) which can act as a deoxynivalenol (DON) antigen has been generated, and its residues Thr102-Ser106 were identified to bind with anti-DON monoclonal antibody by alanine-scanning mutagenesis. Site-saturation mutagenesis was used to analyze the plasticity of five residues and to improve the sensitivity of the N-28-based immunoassay. After mutagenesis, three mutants were selected by phage immunoassay and were sequenced. The half-maximal inhibitory concentrations of the immunoassay based on mutants N-28-T102Y, N-28-V103L, and N-28-Y105F were 24.49 ± 1.0, 51.83 ± 2.5, and 35.65 ± 1.6 ng/mL, respectively, showing the assay was, respectively, 3.2, 1.5, and 2.2 times more sensitive than the wild-type-based assay. The best mutant, N-28-T102Y, was used to develop a competitive phage ELISA to detect DON in cereals with high specificity and accuracy. In addition, the structural properties of N-28-T102Y and N-28 were investigated, revealing that the affinity of N-28-T102Y decreased because of increased steric hindrance with the large side chain. The lower-binding-affinity antigen mimetic may contribute to the improvement of the sensitivity of competitive immunoassays. These results demonstrate that nanobodies would be a favorable tool for engineering. Moreover, our results have laid a solid foundation for site-saturation mutagenesis of antigen-mimicking nanobodies to improve immunoassay sensitivity for small molecules.

Dual-Mode SERS-Fluorescence Immunoassay Using Graphene Quantum Dot Labeling on One-Dimensional Aligned Magnetoplasmonic Nanoparticles.

Science.gov (United States)

Zou, Fengming; Zhou, Hongjian; Tan, Tran Van; Kim, Jeonghyo; Koh, Kwangnak; Lee, Jaebeom

2015-06-10

A novel dual-mode immunoassay based on surface-enhanced Raman scattering (SERS) and fluorescence was designed using graphene quantum dot (GQD) labels to detect a tuberculosis (TB) antigen, CFP-10, via a newly developed sensing platform of linearly aligned magnetoplasmonic (MagPlas) nanoparticles (NPs). The GQDs were excellent bilabeling materials for simultaneous Raman scattering and photoluminescence (PL). The one-dimensional (1D) alignment of MagPlas NPs simplified the immunoassay process and enabled fast, enhanced signal transduction. With a sandwich-type immunoassay using dual-mode nanoprobes, both SERS signals and fluorescence images were recognized in a highly sensitive and selective manner with a detection limit of 0.0511 pg mL(-1).

Discordant Analytical Results Caused by Biotin Interference on Diagnostic Immunoassays in a Pediatric Hospital.

Science.gov (United States)

Ali, Mahesheema; Rajapakshe, Deepthi; Cao, Liyun; Devaraj, Sridevi

2017-09-01

Recent studies have reported that biotin interferes with certain immunoassays. In this study, we evaluated the analytical interference of biotin on immunoassays that use streptavidin-biotin in our pediatric hospital. We tested the effect of different concentrations of biotin (1.5-200 ng/ml) on TSH, Prolactin, Ferritin, CK-MB, β-hCG, Troponin I, LH, FSH, Cortisol, Anti-HAV antibody (IgG and IgM), assays on Ortho Clinical Diagnostic Vitros 5600 Analyzer. Biotin (up to 200 ng/mL) did not significantly affect Troponin I and HAV assays. Biotin (up to 12.5 ng/ml) resulted in biotin >6.25 ng/mL significantly affected TSH (>20% bias) assay. Prolactin was significantly affected even at low levels (Biotin 1.5 ng/mL). Thus, we recommend educating physicians about biotin interference in common immunoassays and adding an electronic disclaimer. © 2017 by the Association of Clinical Scientists, Inc.

340 nm pulsed UV LED system for europium-based time-resolved fluorescence detection of immunoassays

DEFF Research Database (Denmark)

Rodenko, Olga; Fodgaard, Henrik; Tidemand-Lichtenberg, Peter

2016-01-01

We report on the design, development and investigation of an optical system based on UV light emitting diode (LED) excitation at 340 nm for time-resolved fluorescence detection of immunoassays. The system was tested to measure cardiac marker Troponin I with a concentration of 200 ng....../L in immunoassay. The signal-to-noise ratio was comparable to state-of-the-art Xenon flash lamp based unit with equal excitation energy and without overdriving the LED. We performed a comparative study of the flash lamp and the LED based system and discussed temporal, spatial, and spectral features of the LED...... excitation for time-resolved fluorimetry. Optimization of the suggested key parameters of the LED promises significant increase of the signal-to-noise ratio and hence of the sensitivity of immunoassay systems....

340 nm pulsed UV LED system for europium-based time-resolved fluorescence detection of immunoassays.

Science.gov (United States)

Rodenko, Olga; Fodgaard, Henrik; Tidemand-Lichtenberg, Peter; Petersen, Paul Michael; Pedersen, Christian

2016-09-19

We report on the design, development and investigation of an optical system based on UV light emitting diode (LED) excitation at 340 nm for time-resolved fluorescence detection of immunoassays. The system was tested to measure cardiac marker Troponin I with a concentration of 200 ng/L in immunoassay. The signal-to-noise ratio was comparable to state-of-the-art Xenon flash lamp based unit with equal excitation energy and without overdriving the LED. We performed a comparative study of the flash lamp and the LED based system and discussed temporal, spatial, and spectral features of the LED excitation for time-resolved fluorimetry. Optimization of the suggested key parameters of the LED promises significant increase of the signal-to-noise ratio and hence of the sensitivity of immunoassay systems.

Disposable inkjet-printed electrochemical platform for detection of clinically relevant HER-2 breast cancer biomarker.

Science.gov (United States)

Carvajal, Susanita; Fera, Samantha N; Jones, Abby L; Baldo, Thaisa A; Mosa, Islam M; Rusling, James F; Krause, Colleen E

2018-05-01

Rapidly fabricated, disposable sensor platforms hold tremendous promise for point-of-care detection. Here, we present an inexpensive (Receptor 2 (HER-2). Capture antibodies were bound to a chemically modified surface on the WEA and placed into a microfluidic device. A full sandwich immunoassay was constructed following a simultaneous injection of target protein, biotinylated antibody, and polymerized horseradish peroxide labels into the microfluidic device housing the WEA. With an ultra fast assay time, of only 15mins a clinically relevant limit of detection of 12pgmL -1 was achieved. Excellent reproducibility and sensitivity were observed through recovery assays preformed in human serum with recoveries ranging from 76% to 103%. These easily fabricated and scalable electrochemical sensor platforms can be readily adapted for multiplex detection following this rapid assay protocol for cancer diagnostics. Copyright © 2018 Elsevier B.V. All rights reserved.

Magnetic properties dependence on the coupled effects of magnetic fields on the microstructure of as-deposited and post-annealed Co/Ni bilayer thin films

Energy Technology Data Exchange (ETDEWEB)

Franczak, Agnieszka [LISM, Universite de Reims Champagne-Ardenne, BP 1039, 51687 Reims Cedex 2 (France); Department of Materials Science, Katholieke Universiteit Leuven, 3001 Leuven (Belgium); Levesque, Alexandra, E-mail: alexandra.levesque@univ-reims.fr [LISM, Universite de Reims Champagne-Ardenne, BP 1039, 51687 Reims Cedex 2 (France); Coïsson, Marco [Electromagnetism Division, Istituto Nazionale di Ricerca Metrologica, 10135 Torino (Italy); Li, Donggang [LISM, Universite de Reims Champagne-Ardenne, BP 1039, 51687 Reims Cedex 2 (France); Key Laboratory of Electromagnetic Processing of Materials (Ministry of Education), Northeastern University, 110004 Shenyang (China); Barrera, Gabriele [Electromagnetism Division, Istituto Nazionale di Ricerca Metrologica, 10135 Torino (Italy); Università di Torino, Dipartimento di Chimica, 10125 Torino (Italy); Celegato, Federica [Electromagnetism Division, Istituto Nazionale di Ricerca Metrologica, 10135 Torino (Italy); Wang, Qiang [Key Laboratory of Electromagnetic Processing of Materials (Ministry of Education), Northeastern University, 110004 Shenyang (China); Tiberto, Paola [Electromagnetism Division, Istituto Nazionale di Ricerca Metrologica, 10135 Torino (Italy); Chopart, Jean-Paul [LISM, Universite de Reims Champagne-Ardenne, BP 1039, 51687 Reims Cedex 2 (France)

2014-12-15

Magnetic films and multilayers are the focus of much attention motivated mainly by their wide range of applications, such as magnetic data storage devices and sensors. The magnetic multilayer structures are normally prepared through physical means of deposition, as molecular beam epitaxy (MBE) or sputtering. However, there are already examples of materials produced by electrochemical routes, which share with the other deposition techniques a high sensitivity of magnetic and transport properties of the samples on their crystallographic and chemical structure. In addition, electrochemical deposition allows growing structures with high aspect ratio, which are not possible to obtain by MBE deposition followed by lithographic processes. The present work investigates the Co/Ni bilayered nanocrystalline films produced through the temperature-elevated electrochemical deposition, and modified by annealing carried out also under an external magnetic field. The results indicate an increase of the coercive field of deposited Co/Ni bilayers, when the electrodeposition process was conducted under magnetic field of 1 T. The annealing processing caused further remarkable increase of the coercive field of as-prepared bilayers that has been preserved under magnetic annealing conditions. The magnetic properties are discussed in terms of samples microstructure. In as-prepared samples the in-plane magnetization was observed, while high temperature treatment, causing microstructural changes in the film, resulted also in appearance of a small component of magnetization oriented perpendicularly to the films’ plane that could have been observed by MFM analysis. The induced perpendicular magnetization component in the post-annealed samples was a result of the magnetic field applied in the perpendicular direction to the samples’ surface during annealing treatment. - Highlights: • Co deposits were obtained at high electrolyte temperature under applied B-field. • The

Magnetic properties dependence on the coupled effects of magnetic fields on the microstructure of as-deposited and post-annealed Co/Ni bilayer thin films

International Nuclear Information System (INIS)

Franczak, Agnieszka; Levesque, Alexandra; Coïsson, Marco; Li, Donggang; Barrera, Gabriele; Celegato, Federica; Wang, Qiang; Tiberto, Paola; Chopart, Jean-Paul

2014-01-01

Magnetic films and multilayers are the focus of much attention motivated mainly by their wide range of applications, such as magnetic data storage devices and sensors. The magnetic multilayer structures are normally prepared through physical means of deposition, as molecular beam epitaxy (MBE) or sputtering. However, there are already examples of materials produced by electrochemical routes, which share with the other deposition techniques a high sensitivity of magnetic and transport properties of the samples on their crystallographic and chemical structure. In addition, electrochemical deposition allows growing structures with high aspect ratio, which are not possible to obtain by MBE deposition followed by lithographic processes. The present work investigates the Co/Ni bilayered nanocrystalline films produced through the temperature-elevated electrochemical deposition, and modified by annealing carried out also under an external magnetic field. The results indicate an increase of the coercive field of deposited Co/Ni bilayers, when the electrodeposition process was conducted under magnetic field of 1 T. The annealing processing caused further remarkable increase of the coercive field of as-prepared bilayers that has been preserved under magnetic annealing conditions. The magnetic properties are discussed in terms of samples microstructure. In as-prepared samples the in-plane magnetization was observed, while high temperature treatment, causing microstructural changes in the film, resulted also in appearance of a small component of magnetization oriented perpendicularly to the films’ plane that could have been observed by MFM analysis. The induced perpendicular magnetization component in the post-annealed samples was a result of the magnetic field applied in the perpendicular direction to the samples’ surface during annealing treatment. - Highlights: • Co deposits were obtained at high electrolyte temperature under applied B-field. • The

[Automated analyzer of enzyme immunoassay].

Science.gov (United States)

Osawa, S

1995-09-01

Automated analyzers for enzyme immunoassay can be classified by several points of view: the kind of labeled antibodies or enzymes, detection methods, the number of tests per unit time, analytical time and speed per run. In practice, it is important for us consider the several points such as detection limits, the number of tests per unit time, analytical range, and precision. Most of the automated analyzers on the market can randomly access and measure samples. I will describe the recent advance of automated analyzers reviewing their labeling antibodies and enzymes, the detection methods, the number of test per unit time and analytical time and speed per test.

Biosensor immunoassay for flumequine in broiler serum and muscle

NARCIS (Netherlands)

Haasnoot, W.; Gercek, H.; Cazemier, G.; Nielen, M.W.F.

2007-01-01

Flumequine (Flu) is one of the fluoroquinolones most frequently applied for the treatment of broilers in The Netherlands. For the detection of residues of Flu in blood serum of broilers, a biosensor immunoassay (BIA) was developed which was fast (7.5 min per sample) and specific (no cross-reactivity

Chimeric recombinant antibody fragments in cardiac troponin I immunoassay.

Science.gov (United States)

Hyytiä, Heidi; Heikkilä, Taina; Brockmann, Eeva-Christine; Kekki, Henna; Hedberg, Pirjo; Puolakanaho, Tarja; Lövgren, Timo; Pettersson, Kim

2015-03-01

To introduce a novel nanoparticle-based immunoassay for cardiac troponin I (cTnI) utilizing chimeric antibody fragments and to demonstrate that removal of antibody Fc-part and antibody chimerization decrease matrix related interferences. A sandwich-type immunoassay for cTnI based on recombinant chimeric (mouse variable/human constant) antigen binding (cFab) antibodies and intrinsically fluorescent nanoparticles was developed. To test whether using chimeric antibody fragments helps to avoid matrix related interferences, samples (n=39) with known amounts of triglycerides, bilirubin, rheumatoid factor (RF) or human anti-mouse antibodies (HAMAs) were measured with the novel assay, along with a previously published nanoparticle-based research assay with the same antibody epitopes. The limit of detection (LoD) was 3.30ng/L. Within-laboratory precision for 29ng/L and 2819ng/L cTnI were 13.7% and 15.9%, respectively. Regression analysis with Siemens ADVIA Centaur® yielded a slope (95% confidence intervals) of 0.18 (0.17-1.19) and a y-intercept of 1.94 (-1.28-3.91) ng/L. When compared to a previously published nanoparticle-based assay, the novel assay showed substantially reduced interference in the tested interference prone samples, 15.4 vs. 51.3%. A rheumatoid factor containing sample was decreased from 241ng/L to immunoassay for the detection of cTnI and decreased matrix related interferences, thus resulting in a lower number of falsely elevated cTnI-values. Copyright © 2014 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Gliadin Detection in Food by Immunoassay

Science.gov (United States)

Grant, Gordon; Sporns, Peter; Hsieh, Y.-H. Peggy

Immunoassays are very sensitive and efficient tests that are commonly used to identify a specific protein. Examples of applications in the food industry include identification of proteins expressed in genetically modified foods, allergens, or proteins associated with a disease, including celiac disease. This genetic disease is associated with Europeans and affects about one in every 200 people in North America. These individuals react immunologically to wheat proteins, and consequently their own immune systems attack and damage their intestines. This disease can be managed if wheat proteins, specifically "gliadins," are avoided in foods.

Nano-magnetic particles used in biomedicine: core and coating materials.

Science.gov (United States)

Karimi, Z; Karimi, L; Shokrollahi, H

2013-07-01

Magnetic nanoparticles for medical applications have been developed by many researchers. Separation, immunoassay, drug delivery, magnetic resonance imaging and hyperthermia are enhanced by the use of suitable magnetic nanoparticles and coating materials in the form of ferrofluids. Due to their low biocompatibility and low dispersion in water solutions, nanoparticles that are used for biomedical applications require surface treatment. Various kinds of coating materials including organic materials (polymers), inorganic metals (gold, platinum) or metal oxides (aluminum oxide, cobalt oxide) have been attracted during the last few years. Based on the recent advances and the importance of nanomedicine in human life, this paper attempts to give a brief summary on the different ferrite nano-magnetic particles and coatings used in nanomedicine. Copyright © 2013 Elsevier B.V. All rights reserved.

Electrochemical Biosensors - Sensor Principles and Architectures

Directory of Open Access Journals (Sweden)

Erik Reimhult

2008-03-01

Full Text Available Quantification of biological or biochemical processes are of utmost importancefor medical, biological and biotechnological applications. However, converting the biologicalinformation to an easily processed electronic signal is challenging due to the complexity ofconnecting an electronic device directly to a biological environment. Electrochemical biosensorsprovide an attractive means to analyze the content of a biological sample due to thedirect conversion of a biological event to an electronic signal. Over the past decades severalsensing concepts and related devices have been developed. In this review, the most commontraditional techniques, such as cyclic voltammetry, chronoamperometry, chronopotentiometry,impedance spectroscopy, and various field-effect transistor based methods are presented alongwith selected promising novel approaches, such as nanowire or magnetic nanoparticle-basedbiosensing. Additional measurement techniques, which have been shown useful in combinationwith electrochemical detection, are also summarized, such as the electrochemical versionsof surface plasmon resonance, optical waveguide lightmode spectroscopy, ellipsometry,quartz crystal microbalance, and scanning probe microscopy.The signal transduction and the general performance of electrochemical sensors are often determinedby the surface architectures that connect the sensing element to the biological sampleat the nanometer scale. The most common surface modification techniques, the various electrochemicaltransduction mechanisms, and the choice of the recognition receptor moleculesall influence the ultimate sensitivity of the sensor. New nanotechnology-based approaches,such as the use of engineered ion-channels in lipid bilayers, the encapsulation of enzymesinto vesicles, polymersomes, or polyelectrolyte capsules provide additional possibilities forsignal amplification.In particular, this review highlights the importance of the precise control over the

Preparation and influence of pH on the dynamic magnetic property of magnetic FeCoC films

Energy Technology Data Exchange (ETDEWEB)

Feng, Hongmei; Wei, Jinwu; Zhu, Zengtai; Cao, Derang; Liu, Qingfang [Key Laboratory for Magnetism and Magnetic Materials of the Ministry of Education, Lanzhou University, Lanzhou 730000 (China); Wang, Jianbo, E-mail: wangjb@lzu.edu.cn [Key Laboratory for Magnetism and Magnetic Materials of the Ministry of Education, Lanzhou University, Lanzhou 730000 (China); Key Laboratory for Special Function Materials and Structural Design of the Ministry of the Education, Lanzhou University, Lanzhou 730000 (China)

2016-07-01

FeCoC films were successfully prepared by electrochemical deposition method in different citric acid concentrations and pH values. The morphology, structure and magnetic properties were investigated. FeCoC films deposited at different citric acid concentrations have good soft magnetic performance. As the pH value increases from 2.49 to 6.02, the atomic ratio of Fe:Co range from 0.72 to 0.95. The coercivities of the films deposited at different pH values first increase and then decrease with increasing pH. The resonance frequency of the films can be tuned by controlling the pH value, and in an appropriate pH value a wide absorption peak can be obtained. - Highlights: • We have successfully prepared FeCoC soft magnetic films by electrochemical deposition method. • The resonance frequency can be controlled by changing pH value. • A widely absorption peak will be obtained when the pH value is appropriate.

Multiplex dipstick immunoassay for semi-quantitative determination of Fusarium mycotoxins in cereals

International Nuclear Information System (INIS)

Lattanzio, Veronica M.T.; Nivarlet, Noan; Lippolis, Vincenzo; Gatta, Stefania Della; Huet, Anne-Catherine; Delahaut, Philippe; Granier, Benoit; Visconti, Angelo

2012-01-01

Highlights: ► We developed a rapid method based on a multiplex dipstick immunoassay. ► The assay allowed the determination of major Fusarium toxins in wheat, oats, maize. ► We obtained cut off levels close to EU regulatory levels. - Abstract: A multiplex dipstick immunoassay based method for the simultaneous determination of major Fusarium toxins, namely zearalenone, T-2 and HT-2 toxins, deoxynivalenol and fumonisins in wheat, oats and maize has been developed. The dipstick format was based on an indirect competitive approach. Four test lines (mycotoxin–BSA conjugates) and one control line were located on the strip membrane. Labelled antibodies were freeze-dried within the microwell. Two matrix-related sample preparation protocols have been developed for wheat/oats (not containing fumonisins) and maize (containing fumonisins) respectively. The use of a methanol/water mixture for sample preparation allowed recoveries in the range 73–109% for all mycotoxins in all tested cereals, with relative standard deviation less than 10%. The optimized immunoassay was able to detect target mycotoxins at cut off levels equal to 80% of EU maximum permitted levels, i.e. 280, 400, 1400 and 3200 μg kg −1 , respectively, for zearalenone, T-2/HT-2 toxins, deoxynivalenol and fumonisins in maize, and 80, 400 and 1400 μg kg −1 , respectively, for zearalenone, T-2/HT-2 toxins and deoxynivalenol in wheat and oats. Analysis of naturally contaminated samples resulted in a good agreement between multiplex dipstick and validated confirmatory LC–MS/MS. The percentage of false positive results was less than or equal to 13%, whereas no false negative results were obtained. Data on the presence/absence of 6 mycotoxins at levels close to EU regulatory levels were obtained within 30 min. The proposed immunoassay protocol is rapid, inexpensive, easy-to-use and fit for purpose of rapid screening of mycotoxins in cereals.

Activity assays and immunoassays for plasma Renin and prorenin: information provided and precautions necessary for accurate measurement

DEFF Research Database (Denmark)

Campbell, Duncan J; Nussberger, Juerg; Stowasser, Michael

2009-01-01

into focus the differences in information provided by activity assays and immunoassays for renin and prorenin measurement and has drawn attention to the need for precautions to ensure their accurate measurement. CONTENT: Renin activity assays and immunoassays provide related but different information...... provided by these assays and of the precautions necessary to ensure their accuracy....

IFSA: a microfluidic chip-platform for frit-based immunoassay protocols

Science.gov (United States)

Hlawatsch, Nadine; Bangert, Michael; Miethe, Peter; Becker, Holger; Gärtner, Claudia

2013-03-01

Point-of-care diagnostics (POC) is one of the key application fields for lab-on-a-chip devices. While in recent years much of the work has concentrated on integrating complex molecular diagnostic assays onto a microfluidic device, there is a need to also put comparatively simple immunoassay-type protocols on a microfluidic platform. In this paper, we present the development of a microfluidic cartridge using an immunofiltration approach. In this method, the sandwich immunoassay takes place in a porous frit on which the antibodies have immobilized. The device is designed to be able to handle three samples in parallel and up to four analytical targets per sample. In order to meet the critical cost targets for the diagnostic market, the microfluidic chip has been designed and manufactured using high-volume manufacturing technologies in mind. Validation experiments show comparable sensitivities in comparison with conventional immunofiltration kits.

Evaluation of four immunoassays for diagnosis of brucellosis in Cuba

International Nuclear Information System (INIS)

Peraza, C.; Valdes, O.; Fonseca, N.; Garcia, M.; Alvarez, M.; Izquierdo, D.L.

1998-01-01

Four immunoassays (two indirect and two competitive ones) were evaluated by samples from areas free of disease, free by vaccination and affected areas using as reference techniques the Bengal Rose Tests, the Antigen in Buffered Plate Tests and the Complement Fixation Reaction Test. The evaluated samples demonstrated that the competitive assays (ELISAC-1 and ELISAC-2) detected less false positives than the indirect ones (ELISAI-1 and ELISAI-2). Of the competitive ELISAS, version 2 presented better sensitivity and specificity results in affected areas for 95% confidence: 80.9 - 96.9% and 97.5 - 99.4% respectively with positive predictive value in the range of 76 to 94% and negative predictive one between 98.1 and 99.7%. It was concluded that this assay can be used for brucellosis control because it gives higher assurance than the other evaluated immunoassays and it can discriminate infected from vaccinated animals. (author)

Determination of digoxin by enzyme immunoassay and radioimmunoassay

International Nuclear Information System (INIS)

Mueller, H.; Braeuer, H.; Foerster, G.; Reinhardt, M.

1978-01-01

The results of parallel determinations of digoxin in the sera of non selected patients (n=104) by enzyme immunoassay (EMIT.EIA) and radioimmunoassay (J-125 labeled RIA) were compared with each other. The determinations revealed considerably different concentrations; the values determined by EIA were statistical lower (for EIA 1,09+'0,99ng/ml, for RIA 1,34+'1,01ng/ml, p [de

A study using an isotope probe comparing immunoassay with serology in detection of Brucella Abortus antibody

International Nuclear Information System (INIS)

Devlin, J.G.; Redington, F.; Stephenson, M.

1986-01-01

We report a comparison of radio-immunoassay with conventional serology in the detection of brucella abortus antibody from three laboratories. Overall agreement by Chi squared analysis is 5%. There are significant differences between laboratories and a significant number of sero negative suspect sera (from 20% - 60%) were positive by ratio-immunoassay test. We suspect that conventional serology under-reports the incidence of antibody to brucella abortus. (author)

Fast and sensitive detection of enteropathogenic Yersinia by immunoassays.

Science.gov (United States)

Laporte, Jérôme; Savin, Cyril; Lamourette, Patricia; Devilliers, Karine; Volland, Hervé; Carniel, Elisabeth; Créminon, Christophe; Simon, Stéphanie

2015-01-01

Yersinia enterocolitica and Yersinia pseudotuberculosis, the two Yersinia species that are enteropathogenic for humans, are distributed worldwide and frequently cause diarrhea in inhabitants of temperate and cold countries. Y. enterocolitica is a major cause of foodborne disease resulting from consumption of contaminated pork meat and is further associated with substantial economic cost. However, investigation of enteropathogenic Yersinia species is infrequently performed routinely in clinical laboratories because of their specific growth characteristics, which make difficult their isolation from stool samples. Moreover, current isolation procedures are time-consuming and expensive, thus leading to underestimates of the incidence of enteric yersiniosis, inappropriate prescriptions of antibiotic treatments, and unnecessary appendectomies. The main objective of the study was to develop fast, sensitive, specific, and easy-to-use immunoassays, useful for both human and veterinary diagnosis. Monoclonal antibodies (MAbs) directed against Y. enterocolitica bioserotypes 2/O:9 and 4/O:3 and Y. pseudotuberculosis serotypes I and III were produced. Pairs of MAbs were selected by testing their specificity and affinity for enteropathogenic Yersinia and other commonly found enterobacteria. Pairs of MAbs were selected to develop highly sensitive enzyme immunoassays (EIAs) and lateral flow immunoassays (LFIs or dipsticks) convenient for the purpose of rapid diagnosis. The limit of detection of the EIAs ranged from 3.2 × 10(3) CFU/ml to 8.8 × 10(4) CFU/ml for pathogenic serotypes I and III of Y. pseudotuberculosis and pathogenic bioserotypes 2/O:9 and 4/O:3 of Y. enterocolitica and for the LFIs ranged from 10(5) CFU/ml to 10(6) CFU/ml. A similar limit of detection was observed for artificially contaminated human feces. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Amperometric magnetoimmunoassay for the direct detection of tumor necrosis factor alpha biomarker in human serum

Energy Technology Data Exchange (ETDEWEB)

Eletxigerra, U. [Micro-NanoFabrication Unit, IK4-Tekniker, Eibar (Spain); CIC microGUNE, Arrasate-Mondragón (Spain); Martinez-Perdiguero, J. [CIC microGUNE, Arrasate-Mondragón (Spain); Merino, S. [Micro-NanoFabrication Unit, IK4-Tekniker, Eibar (Spain); CIC microGUNE, Arrasate-Mondragón (Spain); Villalonga, R.; Pingarrón, J.M. [Departamento de Química Analítica, Facultad de CC. Químicas, Universidad Complutense de Madrid, Madrid (Spain); Campuzano, S., E-mail: susanacr@quim.ucm.es [Departamento de Química Analítica, Facultad de CC. Químicas, Universidad Complutense de Madrid, Madrid (Spain)

2014-08-01

Highlights: • Electrochemical magnetoimmunosensor for tumor necrosis factor alpha (TNFα) biomarker. • Sensitive and selective detection of TNFα in undiluted serum. • LOD achieved lower than the cut-off value established for relevant illnesses. • Useful and affordable alternative to ELISAs for TNFα determination in serum. - Abstract: An amperometric immunoassay for the determination of tumor necrosis factor alpha (TNFα) protein biomarker in human serum based on the use of magnetic microbeads (MBs) and disposable screen-printed carbon electrodes (SPCEs) has been developed. The specifically modified microbeads were magnetically captured on the working electrode surface and the amperometric responses were measured at −0.20 V (vs. Ag pseudo-reference electrode), upon addition of hydroquinone (HQ) as electron transfer mediator and H{sub 2}O{sub 2} as the enzyme substrate. After a thorough optimization of the assay, extremely low limits of detection were achieved: 2.0 pg mL{sup −1} (36 fM) and 5.8 pg mL{sup −1} (105 fM) for standard solutions and spiked human serum, respectively. The simplicity, robustness and this clinically interesting LOD proved the developed TNFα immunoassay as a good contender for real clinical application.

Amperometric magnetoimmunoassay for the direct detection of tumor necrosis factor alpha biomarker in human serum

International Nuclear Information System (INIS)

Eletxigerra, U.; Martinez-Perdiguero, J.; Merino, S.; Villalonga, R.; Pingarrón, J.M.; Campuzano, S.

2014-01-01

Highlights: • Electrochemical magnetoimmunosensor for tumor necrosis factor alpha (TNFα) biomarker. • Sensitive and selective detection of TNFα in undiluted serum. • LOD achieved lower than the cut-off value established for relevant illnesses. • Useful and affordable alternative to ELISAs for TNFα determination in serum. - Abstract: An amperometric immunoassay for the determination of tumor necrosis factor alpha (TNFα) protein biomarker in human serum based on the use of magnetic microbeads (MBs) and disposable screen-printed carbon electrodes (SPCEs) has been developed. The specifically modified microbeads were magnetically captured on the working electrode surface and the amperometric responses were measured at −0.20 V (vs. Ag pseudo-reference electrode), upon addition of hydroquinone (HQ) as electron transfer mediator and H 2 O 2 as the enzyme substrate. After a thorough optimization of the assay, extremely low limits of detection were achieved: 2.0 pg mL −1 (36 fM) and 5.8 pg mL −1 (105 fM) for standard solutions and spiked human serum, respectively. The simplicity, robustness and this clinically interesting LOD proved the developed TNFα immunoassay as a good contender for real clinical application

Kinase Activity Studied in Living Cells Using an Immunoassay

Science.gov (United States)

Bavec, Aljos?a

2014-01-01

This laboratory exercise demonstrates the use of an immunoassay for studying kinase enzyme activity in living cells. The advantage over the classical method, in which students have to isolate the enzyme from cell material and measure its activity in vitro, is that enzyme activity is modulated and measured in living cells, providing a more…

Comparison of infrared-excited up-converting phosphors and europium nanoparticles as labels in a two-site immunoassay

International Nuclear Information System (INIS)

Ukonaho, Telle; Rantanen, Terhi; Jaemsen, Laura; Kuningas, Katri; Paekkilae, Henna; Loevgren, Timo; Soukka, Tero

2007-01-01

Research in the field of immunoassays and labels used in the detection has been recently focused on particulate reporters, which possess very high specific activity that excludes the label as a sensitivity limiting factor. However, the large size and shape of the particulate labels may produce additional problems to immunoassay performance. The aim of this work was to study with two identical non-competitive two-site immunoassays whether up-converting phosphor (UCP) particles are comparable in performance with europium(III) chelate-dyed nanoparticles as particulate labels. In addition we strived to verify the common assumption of the photostability of up-converting phosphor particles supporting their potential applicability in imaging. Detection limits in two-site immunoassay for free prostate-specific antigen (free-PSA) were 0.53 ng L -1 and 1.3 ng L -1 using two different up-converting phosphors and 0.16 ng L -1 using europium(III) nanoparticle. Large size distribution and non-specific binding of up-converting phosphor particles caused assay variation in low analyte concentrations and limited the analytical detection limit. The non-specific binding was the major factor limiting the analytical sensitivity of the immunoassay. The results suggests the need for nanoscaled and uniformely sized UCP-particles to increace the sensitivity and applicability of up-converting phosphor particles. Anti-Stokes photoluminescence of up-converting phosphor particles did not photobleach when measured repeatedly, on the contrary, the time-resolved fluorescence of europium nanoparticles photobleached relatively rapidly

Immunoassay for Capsular Antigen of Bacillus anthracis Enables Rapid Diagnosis in a Rabbit Model of Inhalational Anthrax.

Directory of Open Access Journals (Sweden)

Marcellene A Gates-Hollingsworth

Full Text Available Inhalational anthrax is a serious biothreat. Effective antibiotic treatment of inhalational anthrax requires early diagnosis; the further the disease has progressed, the less the likelihood for cure. Current means for diagnosis such as blood culture require several days to a result and require advanced laboratory infrastructure. An alternative approach to diagnosis is detection of a Bacillus anthracis antigen that is shed into blood and can be detected by rapid immunoassay. The goal of the study was to evaluate detection of poly-γ-D-glutamic acid (PGA, the capsular antigen of B. anthracis, as a biomarker surrogate for blood culture in a rabbit model of inhalational anthrax. The mean time to a positive blood culture was 26 ± 5.7 h (mean ± standard deviation, whereas the mean time to a positive ELISA was 22 ± 4.2 h; P = 0.005 in comparison with blood culture. A lateral flow immunoassay was constructed for detection of PGA in plasma at concentrations of less than 1 ng PGA/ml. Use of the lateral flow immunoassay for detection of PGA in the rabbit model found that antigen was detected somewhat earlier than the earliest time point at which the blood culture became positive. The low cost, ease of use, and rapid time to result of the lateral flow immunoassay format make an immunoassay for PGA a viable surrogate for blood culture for detection of infection in individuals who have a likelihood of exposure to B. anthracis.

Massively multi-parametric immunoassays using ICPMS

International Nuclear Information System (INIS)

Tanner, S.D.; Ornatsky, O.; Bandura, D.R.; Baranov, V.I.

2009-01-01

The use of stable isotopes as tags in immunoassays, and their determination by ICPMS, is poised to have a huge impact on multi-parametric bioanalysis. A new technology, which we term 'mass cytometry', enables high throughput, highly multiplexed individual cell analysis. Preliminary results for T-cell immunophenotyping in peripheral blood mononuclear cells (PBMC), agonist influence on concomitant phosphorylation pathways, and sub-classification of acute myeloid leukemia patients' samples will be presented. The significance of individual cell analysis is demonstrated by the identification of populations of rogue cells in PBMC samples through the use of multidimensional neural network cluster analysis. (author)

False biochemical diagnosis of hyperthyroidism in streptavidin-biotin-based immunoassays: the problem of biotin intake and related interferences.

Science.gov (United States)

Piketty, Marie-Liesse; Polak, Michel; Flechtner, Isabelle; Gonzales-Briceño, Laura; Souberbielle, Jean-Claude

2017-05-01

Immunoassays are now commonly used for hormone measurement, in high throughput analytical platforms. Immunoassays are generally robust to interference. However, endogenous analytical error may occur in some patients; this may be encountered in biotin supplementation or in the presence of anti-streptavidin antibody, in immunoassays involving streptavidin-biotin interaction. In these cases, the interference may induce both false positive and false negative results, and simulate a seemingly coherent hormonal profile. It is to be feared that this type of errors will be more frequently observed. This review underlines the importance of keeping close interactions between biologists and clinicians to be able to correlate the hormonal assay results with the clinical picture.

Electrochemical activity of thiahelicenes: Structure effects and electrooligomerization ability

International Nuclear Information System (INIS)

Bossi, Alberto; Falciola, Luigi; Graiff, Claudia; Maiorana, Stefano; Rigamonti, Clara; Tiripicchio, Antonio; Licandro, Emanuela; Mussini, Patrizia Romana

2009-01-01

Thiahelicenes are polycondensed heteroaromatic molecules characterized by a chiral helix-like structure including multiple thiophene units, with a lowering effect on the oxidation potentials and a shrinking effect on the band gaps. As a consequence they can be regarded as electrochemically and optically active conducting materials, exhibiting interesting properties under electrical or magnetic polarization, and are under study for non-linear optics (NLO) applications. The present extensive investigation on 11 thiahelicenes with different chain length and functionalization (including the first example of a thiahelicene with perfluorinated alkyl chains) together with the precursor benzodithiophene provides a deep insight on the structure vs. electrochemical activity relationship within this attractive compound class, focusing on both electron transfer (ET) properties and oligomerization ability (hinging on free α positions on terminal thiophene groups).

Targeted selected reaction monitoring mass spectrometric immunoassay for insulin-like growth factor 1.

Directory of Open Access Journals (Sweden)

Eric E Niederkofler

Full Text Available Insulin-like growth factor 1 (IGF1 is an important biomarker of human growth disorders that is routinely analyzed in clinical laboratories. Mass spectrometry-based workflows offer a viable alternative to standard IGF1 immunoassays, which utilize various pre-analytical preparation strategies. In this work we developed an assay that incorporates a novel sample preparation method for dissociating IGF1 from its binding proteins. The workflow also includes an immunoaffinity step using antibody-derivatized pipette tips, followed by elution, trypsin digestion, and LC-MS/MS separation and detection of the signature peptides in a selected reaction monitoring (SRM mode. The resulting quantitative mass spectrometric immunoassay (MSIA exhibited good linearity in the range of 1 to 1,500 ng/mL IGF1, intra- and inter-assay precision with CVs of less than 10%, and lowest limits of detection of 1 ng/mL. The linearity and recovery characteristics of the assay were also established, and the new method compared to a commercially available immunoassay using a large cohort of human serum samples. The IGF1 SRM MSIA is well suited for use in clinical laboratories.

In vitro conditions modify immunoassayability of bovine pituitary prolactin and growth hormone: insights into their secretory granule storage forms

International Nuclear Information System (INIS)

Lorenson, M.Y.

1985-01-01

The amount of immunoassayable intracellular bovine (b) PRL and GH varies depending on treatment conditions. The present studies were designed to characterize the mechanisms involved and to compare immunoassayability of both hormones under similar conditions. Pituitary homogenate and secretory granule hormones displayed both time- and temperature-dependent increases when incubated at pH 10.5 with reduced glutathione. Changes in immunoassayability seem to reflect conversion from poorly immunoactive tissue hormone oligomers to monomeric hormone. The data indicate that oligomeric bPRL is stabilized primarily by intermolecular disulfide bonds, although it is also susceptible to urea, SDS, and EDTA; granule thiols may also influence the conversion to monomer. The storage form of bGH appears to be stabilized differently. Maneuvers demonstrated in these studies to influence immunoassayability correlate very well with their previously established effects on hormone release and secretion, strengthening the likelihood that a functional link exists between assayability and secretion

340 nm pulsed UV LED system for europium-based time-resolved fluorescence detection of immunoassays

OpenAIRE

Rodenko, Olga; Fodgaard, Henrik; Tidemand-Lichtenberg, Peter; Petersen, Paul Michael; Pedersen, Christian

2016-01-01

We report on the design, development and investigation of an optical system based on UV light emitting diode (LED) excitation at 340 nm for time-resolved fluorescence detection of immunoassays. The system was tested to measure cardiac marker Troponin I with a concentration of 200 ng/L in immunoassay. The signal-to-noise ratio was comparable to state-of-the-art Xenon flash lamp based unit with equal excitation energy and without overdriving the LED. We performed a comparative study of the flas...

Electrochemical biosensors

CERN Document Server

Cosnier, Serge

2015-01-01

"This is an excellent book on modern electrochemical biosensors, edited by Professor Cosnier and written by leading international experts. It covers state-of-the-art topics of this important field in a clear and timely manner."-Prof. Joseph Wang, UC San Diego, USA 　"This book covers, in 13 well-illustrated chapters, the potential of electrochemical methods intimately combined with a biological component for the assay of various analytes of biological and environmental interest. Particular attention is devoted to the description of electrochemical microtools in close contact with a biological cell for exocytosis monitoring and to the use of nanomaterials in the electrochemical biosensor architecture for signal improvement. Interestingly, one chapter describes the concept and design of self-powered biosensors derived from biofuel cells. Each topic is reviewed by experts very active in the field. This timely book is well suited for providing a good overview of current research trends devoted to electrochemical...

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.

Electrochemical DNA sandwich assay with a lipase label for attomole detection of DNA

DEFF Research Database (Denmark)

Ferapontova, Elena; Hansen, Majken Nørgaard; Saunders, Aaron Marc

2010-01-01

A fast and sensitive electrochemical lipase-based sandwich hybridization assay for detection of attomole levels of DNA has been developed. A combination of magnetic beads, used for pre-concentration and bioseparation of the analyte with a lipase catalyst label allowed detection of DNA with a limi...

Liquid-liquid phase separation and cluster formation at deposition of metals under inhomogeneous magnetic field

Science.gov (United States)

Gorobets, O. Yu; Gorobets, Yu I.; Rospotniuk, V. P.; Grebinaha, V. I.; Kyba, A. A.

2017-10-01

The formation and dynamic of expansion and deformation of the liquid-liquid interface of an electrolyte at deposition of metals at the surface of the magnetized steel ball is considered in this paper. The electrochemical processes were investigated in an external magnetic field directed at an arbitrary angle to the force of gravity. These processes are accompanied by the formation of effectively paramagnetic clusters of electrochemical products - magnions. Tyndall effect was used for detection of the presence of magnions near the magnetized steel electrode in a solution. The shape of the interface separating the regions with different concentration of magnions, i.e. different magnetic susceptibilities, was described theoretically based on the equation of hydrostatic equilibrium which takes into account magnetic, hydrostatic and osmotic pressures.

Bead-based competitive fluorescence immunoassay for sensitive and rapid diagnosis of cyanotoxin risk in drinking water.

Science.gov (United States)

Yu, Hye-Weon; Jang, Am; Kim, Lan Hee; Kim, Sung-Jo; Kim, In S

2011-09-15

Due to the increased occurrence of cyanobacterial blooms and their toxins in drinking water sources, effective management based on a sensitive and rapid analytical method is in high demand for security of safe water sources and environmental human health. Here, a competitive fluorescence immunoassay of microcystin-LR (MCYST-LR) is developed in an attempt to improve the sensitivity, analysis time, and ease-of-manipulation of analysis. To serve this aim, a bead-based suspension assay was introduced based on two major sensing elements: an antibody-conjugated quantum dot (QD) detection probe and an antigen-immobilized magnetic bead (MB) competitor. The assay was composed of three steps: the competitive immunological reaction of QD detection probes against analytes and MB competitors, magnetic separation and washing, and the optical signal generation of QDs. The fluorescence intensity was found to be inversely proportional to the MCYST-LR concentration. Under optimized conditions, the proposed assay performed well for the identification and quantitative analysis of MCYST-LR (within 30 min in the range of 0.42-25 μg/L, with a limit of detection of 0.03 μg/L). It is thus expected that this enhanced assay can contribute both to the sensitive and rapid diagnosis of cyanotoxin risk in drinking water and effective management procedures.

Octa- and nonanuclear nickel(II) polyoxometalate clusters: synthesis and electrochemical and magnetic characterizations.

Science.gov (United States)

Pichon, Céline; Mialane, Pierre; Dolbecq, Anne; Marrot, Jérôme; Rivière, Eric; Bassil, Bassem S; Kortz, Ulrich; Keita, Bineta; Nadjo, Louis; Sécheresse, Francis

2008-12-01

Three high-nuclearity NiII-substituted polyoxometalate compounds functionalized by exogenous ligands have been synthesized and characterized. The octanuclear complexes in Na15[Na{(A-R-SiW9O34)Ni4(CH3COO)3(OH)3}2] . 4NaCl . 36H2O (1) and Na15[Na{(A-R-SiW9O34)Ni4(CH3COO)3(OH)2(N3)}2] . 32H2O (2) can be described as two {Ni4} subunits connected via a {Na(CH3COO)6} group, with the acetato ligands also ensuring in each subunit the connection between the paramagnetic centers. In 2, two azido groups replace two of the six mu-hydroxo ligands present in 1. The nonanuclear complex K7Na7[(A-R-SiW9O34)2Ni9(OH)6(H2O)6(CO3)3] . 42H2O (3) exhibits a double cubanestructure with two [(A-R-SiW9O34)Ni4(OH)3]5- subunits linked by three carbonato ligands. A ninth NiII center connected to one subunit via a carbonato ligand and a O=W group completes this asymmetric polyoxometalate.Electronic spectroscopy and electrochemical studies indicate that, while compounds 1-3 decompose in a pure aqueous medium, these complexes are very stable in a pH 6 acetate medium. The cyclic voltammetry pattern of each complex is constituted by a first eight-electron reduction wave followed by a second large-current intensity wave. The characteristics of the first waves of the complexes are clearly distinct from those obtained for their lacunary precursor [A-R-SiW9O34]10-, a feature that is due to the Ni centers in the complexes. Such observations of electroactive, stable, and highly nickel-rich polyoxometalates are not common. Measurements of the magnetic susceptibility revealed the occurrence of concomitant ferromagnetic and antiferromagnetic interactions in 1 and 3.For both of these compounds, the extension of the magnetic exchange has been determined by means of a spin Hamiltonian with three and four J constants, respectively.

Electrochemical energy generation

International Nuclear Information System (INIS)

Kreysa, G.; Juettner, K.

1993-01-01

The proceedings encompass 40 conference papers belonging to the following subject areas: Baseline and review papers; electrochemical fuel cells; batteries: Primary and secondary cells; electrochemical, regenerative systems for energy conversion; electrochemical hydrogen generation; electrochemistry for nuclear power plant; electrochemistry for spent nuclear fuel reprocessing; energy efficiency in electrochemical processes. There is an annex listing the authors and titles of the poster session, and compacts of the posters can be obtained from the office of the Gesellschaft Deutscher Chemiker, Abteilung Tagungen. (MM) [de

Correlations between calcineurin phosphatase inhibition and cyclosporine metabolites concentrations in kidney transplant recipients: Implications for immunoassays

DEFF Research Database (Denmark)

Jørgensen, Kaj Anker; Karamperis, Nikolaos; Koefoed-Nielsen, Pernille Bundgaard

2006-01-01

by inhibiting the enzyme calcineurin phosphatase. Determination of the enzyme's activity is one of the most promising pharmacodynamic markers. It is unknown how calcineurin phosphatase inhibition correlates with various cyclosporine monitoring assays and what is the potential impact of metabolites...... by the enzyme multiplied immunoassay technique (EMIT) and by the polyclonal fluorescence polarization immunoassay (pFPIA). Calcineurin phosphatase activity was measured by its ability to dephosphorylate a previously phosphorylated 19-amino acid peptide. We found that calcineurin phosphatase inhibition...

Correlations between calcineurin phosphatase inhibition and cyclosporine metabolites concentrations in kidney transplant recipients: implications for immunoassays

DEFF Research Database (Denmark)

Karamperis, N; Koefoed-Nielsen, PB; Brahe, P

2006-01-01

by inhibiting the enzyme calcineurin phosphatase. Determination of the enzyme's activity is one of the most promising pharmacodynamic markers. It is unknown how calcineurin phosphatase inhibition correlates with various cyclosporine monitoring assays and what is the potential impact of metabolites...... by the enzyme multiplied immunoassay technique (EMIT) and by the polyclonal fluorescence polarization immunoassay (pFPIA). Calcineurin phosphatase activity was measured by its ability to dephosphorylate a previously phosphorylated 19-amino acid peptide. We found that calcineurin phosphatase inhibition...

Multiplex dipstick immunoassay for semi-quantitative determination of Fusarium mycotoxins in cereals

Energy Technology Data Exchange (ETDEWEB)

Lattanzio, Veronica M.T., E-mail: veronica.lattanzio@ispa.cnr.it [National Research Council of Italy, Institute of Sciences of Food Production (ISPA-CNR), Via Amendola 122/O, 70126 Bari (Italy); Nivarlet, Noan [UNISENSOR S.A., Zoning industriel du Dossay, Rue du Dossay no 3, B-4020 Liege (Belgium); Lippolis, Vincenzo; Gatta, Stefania Della [National Research Council of Italy, Institute of Sciences of Food Production (ISPA-CNR), Via Amendola 122/O, 70126 Bari (Italy); Huet, Anne-Catherine; Delahaut, Philippe [Centre d' Economie Rurale (CER Groupe), Rue du Point du Jour no 8, B-6900 Marloie (Belgium); Granier, Benoit [UNISENSOR S.A., Zoning industriel du Dossay, Rue du Dossay no 3, B-4020 Liege (Belgium); Visconti, Angelo [National Research Council of Italy, Institute of Sciences of Food Production (ISPA-CNR), Via Amendola 122/O, 70126 Bari (Italy)

2012-03-09

Highlights: Black-Right-Pointing-Pointer We developed a rapid method based on a multiplex dipstick immunoassay. Black-Right-Pointing-Pointer The assay allowed the determination of major Fusarium toxins in wheat, oats, maize. Black-Right-Pointing-Pointer We obtained cut off levels close to EU regulatory levels. - Abstract: A multiplex dipstick immunoassay based method for the simultaneous determination of major Fusarium toxins, namely zearalenone, T-2 and HT-2 toxins, deoxynivalenol and fumonisins in wheat, oats and maize has been developed. The dipstick format was based on an indirect competitive approach. Four test lines (mycotoxin-BSA conjugates) and one control line were located on the strip membrane. Labelled antibodies were freeze-dried within the microwell. Two matrix-related sample preparation protocols have been developed for wheat/oats (not containing fumonisins) and maize (containing fumonisins) respectively. The use of a methanol/water mixture for sample preparation allowed recoveries in the range 73-109% for all mycotoxins in all tested cereals, with relative standard deviation less than 10%. The optimized immunoassay was able to detect target mycotoxins at cut off levels equal to 80% of EU maximum permitted levels, i.e. 280, 400, 1400 and 3200 {mu}g kg{sup -1}, respectively, for zearalenone, T-2/HT-2 toxins, deoxynivalenol and fumonisins in maize, and 80, 400 and 1400 {mu}g kg{sup -1}, respectively, for zearalenone, T-2/HT-2 toxins and deoxynivalenol in wheat and oats. Analysis of naturally contaminated samples resulted in a good agreement between multiplex dipstick and validated confirmatory LC-MS/MS. The percentage of false positive results was less than or equal to 13%, whereas no false negative results were obtained. Data on the presence/absence of 6 mycotoxins at levels close to EU regulatory levels were obtained within 30 min. The proposed immunoassay protocol is rapid, inexpensive, easy-to-use and fit for purpose of rapid screening of mycotoxins

Aequorin fusion proteins as bioluminescent tracers for competitive immunoassays

Science.gov (United States)

Mirasoli, Mara; Michelini, Elisa; Deo, Sapna K.; Dikici, Emre; Roda, Aldo; Daunert, Sylvia

2004-06-01

The use of bio- and chemiluminescence for the development of quantitative binding assays offers undoubted advantages over other detection systems, such as spectrophotometry, fluorescence, or radioactivity. Indeed, bio- and chemiluminescence detection provides similar, or even better, sensitivity and detectability than radioisotopes, while avoiding the problems of health hazards, waste disposal, and instability associated with the use of radioisotopes. Among bioluminescent labels, the calcium-activated photoprotein aequorin, originally isolated from Aequorea victoria and today available as a recombinant product, is characterized by very high detectability, down to attomole levels. It has been used as a bioluminescent label for developing a variety of highly sensitive immunoassays, using various analyte-aequorin conjugation strategies. When the analyte is a protein or a peptide, genetic engineering techniques can be used to produce protein fusions where the analyte is in-frame fused with aequorin, thus producing homogeneous one-to-one conjugation products, available in virtually unlimited amount. Various assays were developed using this strategy: a short review of the most interesting applications is presented, as well as the cloning, purification and initial characterization of an endothelin-1-aequorin conjugate suitable for developing a competitive immunoassay for endothelin-1, a potent vasoconstrictor peptide, involved in hypertension.

Trace-level mercury ion (Hg2+) analysis in aqueous sample based on solid-phase extraction followed by microfluidic immunoassay.

Science.gov (United States)

Date, Yasumoto; Aota, Arata; Terakado, Shingo; Sasaki, Kazuhiro; Matsumoto, Norio; Watanabe, Yoshitomo; Matsue, Tomokazu; Ohmura, Naoya

2013-01-02

Mercury is considered the most important heavy-metal pollutant, because of the likelihood of bioaccumulation and toxicity. Monitoring widespread ionic mercury (Hg(2+)) contamination requires high-throughput and cost-effective methods to screen large numbers of environmental samples. In this study, we developed a simple and sensitive analysis for Hg(2+) in environmental aqueous samples by combining a microfluidic immunoassay and solid-phase extraction (SPE). Using a microfluidic platform, an ultrasensitive Hg(2+) immunoassay, which yields results within only 10 min and with a lower detection limit (LOD) of 0.13 μg/L, was developed. To allow application of the developed immunoassay to actual environmental aqueous samples, we developed an ion-exchange resin (IER)-based SPE for selective Hg(2+) extraction from an ion mixture. When using optimized SPE conditions, followed by the microfluidic immunoassay, the LOD of the assay was 0.83 μg/L, which satisfied the guideline values for drinking water suggested by the United States Environmental Protection Agency (USEPA) (2 μg/L; total mercury), and the World Health Organisation (WHO) (6 μg/L; inorganic mercury). Actual water samples, including tap water, mineral water, and river water, which had been spiked with trace levels of Hg(2+), were well-analyzed by SPE, followed by microfluidic Hg(2+) immunoassay, and the results agreed with those obtained from reduction vaporizing-atomic adsorption spectroscopy.

Horseradish peroxidase-loaded nanospheres attached to hollow gold nanoparticles as signal enhancers in an ultrasensitive immunoassay for alpha-fetoprotein

International Nuclear Information System (INIS)

Li, Ya; Yuan, Ruo; Chai, Yaqin; Zhuo, Ying; Su, Huilan; Zhang, Yuxia

2014-01-01

We report on a novel electrochemical signal amplification strategy for use in immunoassays. The highly responsive immunoelectrode was constructed in the following way: (1) The surface of a gold electrode was covered with a layer single-walled carbon nanotubes dispersed in chitosane functionalized with L-cysteine; (2) Gold nanoparticles containing protein A and anti-alpha-fetoprotein (anti-AFP) were then covalently attached to the surface via the thiol groups of the chitosane. The electrode is then exposed to the analyte (AFP) which then is bound by the antibody. In the next step, a gold-conjugated secondary antibody is added that was prepared in the following way: (1) Horseradish peroxidase was crosslinked and the resulting spheres were coated with hollow gold nanoparticles (hollow Au-NPs) to give nanospheres of ∼100 nm in diameter. (2) These were the coated with thionine and, in a last step, with secondary antibody. The use of these materials has several attractive features: The HRP-NPs functionalized with hollow Au-NPs possess a large surface area that can load the large amount of secondary antibody. Thionine (Thi) is highly redox active and improves the intensity of the signal. Carbon nanotubes were used because they possess an excellent electron transfer rate and large surface area. Following incubation of the modified electrode (a) with a sample containing AFP, (b) then with the secondary antibody, and (c) with washing buffer, the electrode is placed in a solution containing H 2 O 2 . The HRP in the smart secondary antibody causes the catalytic decomposition of H 2 O 2 and the results in an electrical current that is linearly related to the concentration of AFP in the 0.025 to 5.0 ng mL −1 concentration range. The detection limit for AFP is as low as 8.3 pg mL −1 . We believe that this novel kind of immunoassay represents a promising tool for use in sensitive clinical assays. (author)

An enzyme immunoassay to quantify neurofilament light chain in cerebrospinal fluid.

NARCIS (Netherlands)

Geel, W.J.A. van; Rosengren, L.E.; Verbeek, M.M.

2005-01-01

Neurofilament light chain is a component of the axonal cytoskeleton. The concentration of the neurofilament light chain in cerebrospinal fluid may reflect axonal damage or the extent of white matter damage. In this study we describe a sensitive immunoassay for the detection of neurofilament light

IMMUNOASSAY METHOD FOR THE DETERMINATION OF PENTACHLOROPHENOL IN SOIL AND SEDIMENT

Science.gov (United States)

The journal article describes the use of a prototype immunoassay method for the determination of pentacholorphenol (PCP) in soil and sediment. PCP was used as a pesticide and wood preservative and is not currently available to the general public. The paper stresses the importan...

Immunoassay for thymopoietin

International Nuclear Information System (INIS)

Goldstein, G.

1979-01-01

The patent describes the development of a radio-immunoassay for thymopoietin in biological samples. The method of raising antibodies to this polypeptide hormone is described. This is achieved by injecting a host animal with an antigen consisting of thymopoietin covalently bonded by glutaraldehyde to a carrier protein such as bovine serum albumin and equine globulin. Different methods of radiolabelling thymopoietin with 125 I for use as the tracer antigen are described. The Bolton-Hunter procedure was preferred to the chloramine-T method since direct iodination of the tyrosyl moieties of thymopoietin resulted in some loss of immunoreactivity. Systems for separating the antigen-antibody complex and unbound antigen are compared. Binding-inhibition curves for unlabelled thymopoietin in the assay employing polyethylene glycol separation showed a sensitivity of 5 ng thymopoietin/ml. However, using the double antibody or dextran coated charcoal separation techniques, the sensitivity of thymopoietin was 0.1 ng/ml. Thus these latter two procedures are thus especially suitable for measuring thymopoietin levels in serum or plasma samples. The assay was shown to be specific for thymopoietin, no significant displacement being produced by control polypeptides. (U.K.)

Buffered Electrochemical Polishing of Niobium

Energy Technology Data Exchange (ETDEWEB)

Ciovati, Gianluigi [Thomas Jefferson National Accelerator Facility (TJNAF), Newport News, VA (United States); Tian, Hui [Thomas Jefferson National Accelerator Facility (TJNAF), Newport News, VA (United States); College of William and Mary, Williamsburg, VA (United States); Corcoran, Sean [Virginia Polytechnic Inst. and State Univ. (Virginia Tech), Blacksburg, VA (United States)

2011-03-01

The standard preparation of superconducting radio-frequency (SRF) cavities made of pure niobium include the removal of a 'damaged' surface layer, by buffered chemical polishing (BCP) or electropolishing (EP), after the cavities are formed. The performance of the cavities is characterized by a sharp degradation of the quality factor when the surface magnetic field exceeds about 90 mT, a phenomenon referred to as 'Q-drop.' In cavities made of polycrystalline fine grain (ASTM 5) niobium, the Q-drop can be significantly reduced by a low-temperature (? 120 °C) 'in-situ' baking of the cavity if the chemical treatment was EP rather than BCP. As part of the effort to understand this phenomenon, we investigated the effect of introducing a polarization potential during buffered chemical polishing, creating a process which is between the standard BCP and EP. While preliminary results on the application of this process to Nb cavities have been previously reported, in this contribution we focus on the characterization of this novel electrochemical process by measuring polarization curves, etching rates, surface finish, electrochemical impedance and the effects of temperature and electrolyte composition. In particular, it is shown that the anodic potential of Nb during BCP reduces the etching rate and improves the surface finish.

The annealing temperature dependences of microstructures and magnetic properties in electro-chemical deposited CoNiFe thin films

International Nuclear Information System (INIS)

Suharyadi, Edi; Riyanto, Agus; Abraha, Kamsul

2016-01-01

CoNiFe thin films with various compositions had been successfully fabricated using electro-chemical deposition method. The crystal structure of Co_6_5Ni_1_5Fe_2_0, Co_6_2Ni_1_5Fe_2_3, and Co_5_5Ni_1_5Fe_3_0 thin films was fcc, bcc-fcc mix, and bcc, respectively. The difference crystal structure results the difference in magnetic properties. The saturation magnetic flux density (Bs) of Co_6_5Ni_1_5Fe_2_0, Co_6_2Ni_1_5Fe_2_3, and Co_5_5Ni_1_5Fe_3_0 thin films was 1.89 T, 1.93 T, and 2.05 T, respectively. An optimal annealing temperature was determined for controlling the microstructure and magnetic properties of CoNiFe thin films. Depending on annealing temperature, the ratio of bcc and fcc structure varied without changing the film composition. By annealing at temperature of T ≥ 350°C, the intensity ratio of X-ray diffraction peaks for bcc(110) to fcc(111) increased. The increase of phase ratio of bcc(110) to fcc(111) caused the increase of Bs, from 1.89 T to 1.95 T. Coercivity (Hc) also increased after annealing, from 2.6 Oe to 18.6 Oe for fcc phase thin films, from 2.0 Oe to 12.0 Oe for fcc-bcc mix phase thin films, and 7.8 Oe to 8 Oe for bcc phase thin films. The changing crystal structures during annealing process indicated that the thermal treatment at high temperature cause the changing crystallinity and atomic displacement. The TEM bright-field images with corresponding selected-area electron diffraction (SAED) patterns showed that there are strongly effects of thermal annealing on the size of fcc and bcc phase crystalline grain as described by size of individual spot and discontinuous rings. The size of crystalline grains increased by thermal annealing. The evolution of bcc and fcc structures of CoNiFe during annealing is though to be responsible for the change of magnetic properties.

Serum sample containing endogenous antibodies interfering with multiple hormone immunoassays. Laboratory strategies to detect interference

Directory of Open Access Journals (Sweden)

Elena García-González

2016-04-01

Full Text Available Objectives: Endogenous antibodies (EA may interfere with immunoassays, causing erroneous results for hormone analyses. As (in most cases this interference arises from the assay format and most immunoassays, even from different manufacturers, are constructed in a similar way, it is possible for a single type of EA to interfere with different immunoassays. Here we describe the case of a patient whose serum sample contains EA that interfere several hormones tests. We also discuss the strategies deployed to detect interference. Subjects and methods: Over a period of four years, a 30-year-old man was subjected to a plethora of laboratory and imaging diagnostic procedures as a consequence of elevated hormone results, mainly of pituitary origin, which did not correlate with the overall clinical picture. Results: Once analytical interference was suspected, the best laboratory approaches to investigate it were sample reanalysis on an alternative platform and sample incubation with antibody blocking tubes. Construction of an in-house ‘nonsense’ sandwich assay was also a valuable strategy to confirm interference. In contrast, serial sample dilutions were of no value in our case, while polyethylene glycol (PEG precipitation gave inconclusive results, probably due to the use of inappropriate PEG concentrations for several of the tests assayed. Conclusions: Clinicians and laboratorians must be aware of the drawbacks of immunometric assays, and alert to the possibility of EA interference when results do not fit the clinical pattern. Keywords: Endogenous antibodies, Immunoassay, Interference, Pituitary hormones, Case report

Functional Polymers in Protein Detection Platforms: Optical, Electrochemical, Electrical, Mass-Sensitive, and Magnetic Biosensors

Directory of Open Access Journals (Sweden)

Jong-in Hahm

2011-03-01

Full Text Available The rapidly growing field of proteomics and related applied sectors in the life sciences demands convenient methodologies for detecting and measuring the levels of specific proteins as well as for screening and analyzing for interacting protein systems. Materials utilized for such protein detection and measurement platforms should meet particular specifications which include ease-of-mass manufacture, biological stability, chemical functionality, cost effectiveness, and portability. Polymers can satisfy many of these requirements and are often considered as choice materials in various biological detection platforms. Therefore, tremendous research efforts have been made for developing new polymers both in macroscopic and nanoscopic length scales as well as applying existing polymeric materials for protein measurements. In this review article, both conventional and alternative techniques for protein detection are overviewed while focusing on the use of various polymeric materials in different protein sensing technologies. Among many available detection mechanisms, most common approaches such as optical, electrochemical, electrical, mass-sensitive, and magnetic methods are comprehensively discussed in this article. Desired properties of polymers exploited for each type of protein detection approach are summarized. Current challenges associated with the application of polymeric materials are examined in each protein detection category. Difficulties facing both quantitative and qualitative protein measurements are also identified. The latest efforts on the development and evaluation of nanoscale polymeric systems for improved protein detection are also discussed from the standpoint of quantitative and qualitative measurements. Finally, future research directions towards further advancements in the field are considered.

Metal-doped inorganic nanoparticles for multiplex detection of biomarkers by a sandwich-type ICP-MS immunoassay.

Science.gov (United States)

Ko, Jung Aa; Lim, H B

2016-09-28

Metal-doped inorganic nanoparticles were synthesized for the multiplex detection of biomarkers by a sandwich-type inductively coupled plasma mass spectrometry (ICP-MS) immunoassay. The synthesized Cs-doped multicore magnetic nanoparticles (MMNPs) were used not only for magnetic extraction of targets but also for ratiometric measurement in ICP-MS. In addition, three different metal/dye-doped silica nanoparticles (SNPs) were synthesized as probes for multiplex detection: Y/RhBITC (rhodamine B isothiocyanate)-doped SNPs for CRP (cardiovascular disease), Cd/RhBITC-doped SNPs for AFP (tumor), and Au/5(6)-XRITC (X-rhodamine-5-(and-6)-isothiocyanate)-doped SNPs for NSE (heart disease). For quantification, the doped metals of SNPs were measured by ICP-MS and then the signal ratio to Cs of MMNPs was plotted with respect to the concentration of targets by a ratiometry. Limits of detection (LOD) of 0.35 ng/mL to 77 ng mL(-1) and recoveries of 83%-125% were obtained for serum samples spiked with the biomarkers. Since no sample treatment was necessary prior to the extraction, the proposed method provided short analysis time and convenience for the multiplex determination of biomarkers, which will be valuable for clinical application. Copyright © 2016 Elsevier B.V. All rights reserved.

On-Chip Immunoassay for Determination of Urinary Albumin

OpenAIRE

Laiwattanapaisal, Wanida; Songjaroen, Temsiri; Maturos, Thitima; Lomas, Tanom; Sappat, Assawapong; Tuantranont, Adisorn

2009-01-01

An immunoassay performed on a portable microfluidic device was evaluated for the determination of urinary albumin. An increase in absorbance at 500 nm resulting from immunoagglutination was monitored directly on the poly(dimethylsiloxane) (PDMS) microchip using a portable miniature fibre-optic spectrometer. A calibration curve was linear up to 10 mg L–1 (r2 = 0.993), with a detection limit of 0.81 mg L–1 (S/N = 3). The proposed system showed good precision, with relative standard deviations (...

Magnetic Bead and Fluorescent Silica Nanoparticles Based Optical Immunodetection of Staphylococcal Enterotoxin B (SEB in Bottled Water

Directory of Open Access Journals (Sweden)

Shiva K. RASTOGI

2009-10-01

Full Text Available Staphylococcal enterotoxins (SEs are a major cause of food-borne diseases, most commonly SEs assayed immunologically with ELISA. An immunoassay based on fluorescein dye doped silica dioxide nanoparticles (F-SiNPs and magnetic bead (MB is described here for the detection of staphylococcal enterotoxin B (SEB. F-SiNPs have unique optical properties which make them attractive for biosensing. The water-in-oil (W/O reverse microemulsion method was used for the synthesis of F-SiNPs (~ 95 nm of diameter. The F-SiNPs were characterized using SEM, TEM and FTIR spectroscopy. The detection of SEB is preformed in PBS buffer, and bottled drinking water using sandwich immunoassay format. Target analytes were captured using MBs modified with the antigen-specific “capture” antibody, and detected using F-SiNP labeled secondary antigen-specific antibody. We report a limit of detection down to 1 ng/mL SEB spiked sample in less than 2 hr assay time using fluorocount method. This study demonstrates the bio warfare agent SEB capture by magnetic beads and detection using F-SiNPs.

Immunoassay: Principles, development and potential applications in the applied plant sciences

Energy Technology Data Exchange (ETDEWEB)

Hofman, P J

1986-02-01

The article briefly discusses the general principles of, and the methods involved in, immunoassay, and their development. Emplasis is placed on radioimmunoassay (RIA) and to a lesser extent, enzyme-linked immunosorbent assay (ELISA). The practical applications, with special reference to the citrus and subtropical fruit industries are discussed.

Design of novel hybrid organic-inorganic nanostructured biomaterials for immunoassay applications

Energy Technology Data Exchange (ETDEWEB)

Andrade, G [Department of Microbiology, Institute of Biological Sciences, Federal University of Minas Gerais, PO Box 486, 31270.901, Belo Horizonte, MG (Brazil); Barbosa-Stancioli, E F [Department of Microbiology, Institute of Biological Sciences, Federal University of Minas Gerais, PO Box 486, 31270.901, Belo Horizonte, MG (Brazil); Piscitelli Mansur, A A [Department of Metallurgical and Materials Engineering, Biomaterials and Tissue Engineering Laboratory, Federal University of Minas Gerais, Belo Horizonte, MG (Brazil); Vasconcelos, W L [Department of Metallurgical and Materials Engineering, Biomaterials and Tissue Engineering Laboratory, Federal University of Minas Gerais, Belo Horizonte, MG (Brazil); Mansur, H S [Department of Metallurgical and Materials Engineering, Biomaterials and Tissue Engineering Laboratory, Federal University of Minas Gerais, Belo Horizonte, MG (Brazil)

2006-12-01

The purpose of this study was to develop novel hybrid organic-inorganic materials based on poly(vinyl alcohol) (PVA) polymer chemically crosslinked network to be tested as solid support on bovine herpesvirus immunoassay. Hybrids were synthesized by reacting PVA with three different alkoxysilanes modifying chemical groups: tetraethoxysilane (TEOS), 3-mercaptopropyltrimethoxysilane (MPTMS) and 3-glycidoxypropyltrimethoxysilane (GPTMS). PVA-derived hybrids were also modified by chemically crosslinking with glutaraldehyde (GA) during the synthesis reaction. In order to investigate the structure in the nanometer-scale, PVA-derived hybrids were characterized by using small-angle x-ray scattering synchrotron radiation (SAXS) and x-ray diffraction (XRD). PVA hybrids' chemical functionalities and their interaction with herpesviruses were also characterized by Fourier transform infrared spectroscopy (FTIR). The bioactivity assays were tested through enzyme linked immunosorbent assay (ELISA). SAXS results have indicated nano-ordered disperse domains for PVA hybrids with different x-ray scattering patterns for PVA polymer and PVA-derived hybrids. FTIR spectra have shown major vibration bands associated with organic-inorganic chemical groups present in the PVA, PVA-derived by silane modifier and PVA chemically crosslinked by GA. The immunoassay results have shown that PVA hybrids with chemically functionalized structures regulated to some extent the specific bioimmobilization of herpesvirus onto solid phase. We think that it is due to the overall balance of forces associated with van der Waals interaction, hydrophilic and hydrophobic forces and steric hindrance acting at the surface. PVA and PVA-derived hybrid materials were successfully produced with GA crosslinking in a nanometer-scale network. Also, such a PVA-based material could be advantageously used in immunoassays with enhanced specificity for diagnosis.

Design of novel hybrid organic-inorganic nanostructured biomaterials for immunoassay applications

Energy Technology Data Exchange (ETDEWEB)

Andrade, G [Department of Microbiology, Institute of Biological Sciences, Federal University of Minas Gerais, PO Box 486, 31270.901, Belo Horizonte, MG (Brazil); Barbosa-Stancioli, E F [Department of Microbiology, Institute of Biological Sciences, Federal University of Minas Gerais, PO Box 486, 31270.901, Belo Horizonte, MG (Brazil); Piscitelli Mansur, A A [Department of Metallurgical and Materials Engineering, Biomaterials and Tissue Engineering Laboratory, Federal University of Minas Gerais, Belo Horizonte, MG (Brazil); Vasconcelos, W L [Department of Metallurgical and Materials Engineering, Biomaterials and Tissue Engineering Laboratory, Federal University of Minas Gerais, Belo Horizonte, MG (Brazil); Mansur, H S [Department of Metallurgical and Materials Engineering, Biomaterials and Tissue Engineering Laboratory, Federal University of Minas Gerais, Belo Horizonte, MG (Brazil)

2006-12-01

The purpose of this study was to develop novel hybrid organic-inorganic materials based on poly(vinyl alcohol) (PVA) polymer chemically crosslinked network to be tested as solid support on bovine herpesvirus immunoassay. Hybrids were synthesized by reacting PVA with three different alkoxysilanes modifying chemical groups: tetraethoxysilane (TEOS), 3-mercaptopropyltrimethoxysilane (MPTMS) and 3-glycidoxypropyltrimethoxysilane (GPTMS). PVA-derived hybrids were also modified by chemically crosslinking with glutaraldehyde (GA) during the synthesis reaction. In order to investigate the structure in the nanometer-scale, PVA-derived hybrids were characterized by using small-angle x-ray scattering synchrotron radiation (SAXS) and x-ray diffraction (XRD). PVA hybrids' chemical functionalities and their interaction with herpesviruses were also characterized by Fourier transform infrared spectroscopy (FTIR). The bioactivity assays were tested through enzyme linked immunosorbent assay (ELISA). SAXS results have indicated nano-ordered disperse domains for PVA hybrids with different x-ray scattering patterns for PVA polymer and PVA-derived hybrids. FTIR spectra have shown major vibration bands associated with organic-inorganic chemical groups present in the PVA, PVA-derived by silane modifier and PVA chemically crosslinked by GA. The immunoassay results have shown that PVA hybrids with chemically functionalized structures regulated to some extent the specific bioimmobilization of herpesvirus onto solid phase. We think that it is due to the overall balance of forces associated with van der Waals interaction, hydrophilic and hydrophobic forces and steric hindrance acting at the surface. PVA and PVA-derived hybrid materials were successfully produced with GA crosslinking in a nanometer-scale network. Also, such a PVA-based material could be advantageously used in immunoassays with enhanced specificity for diagnosis.

Development of a prototype lateral flow immunoassay (LFI for the rapid diagnosis of melioidosis.

Directory of Open Access Journals (Sweden)

Raymond L Houghton

2014-03-01

Full Text Available Burkholderia pseudomallei is a soil-dwelling bacterium and the causative agent of melioidosis. Isolation of B. pseudomallei from clinical samples is the "gold standard" for the diagnosis of melioidosis; results can take 3-7 days to produce. Alternatively, antibody-based tests have low specificity due to a high percentage of seropositive individuals in endemic areas. There is a clear need to develop a rapid point-of-care antigen detection assay for the diagnosis of melioidosis. Previously, we employed In vivo Microbial Antigen Discovery (InMAD to identify potential B. pseudomallei diagnostic biomarkers. The B. pseudomallei capsular polysaccharide (CPS and numerous protein antigens were identified as potential candidates. Here, we describe the development of a diagnostic immunoassay based on the detection of CPS. Following production of a CPS-specific monoclonal antibody (mAb, an antigen-capture immunoassay was developed to determine the concentration of CPS within a panel of melioidosis patient serum and urine samples. The same mAb was used to produce a prototype Active Melioidosis Detect Lateral Flow Immunoassay (AMD LFI; the limit of detection of the LFI for CPS is comparable to the antigen-capture immunoassay (∼0.2 ng/ml. The analytical reactivity (inclusivity of the AMD LFI was 98.7% (76/77 when tested against a large panel of B. pseudomallei isolates. Analytical specificity (cross-reactivity testing determined that 97.2% of B. pseudomallei near neighbor species (35/36 were not reactive. The non-reactive B. pseudomallei strain and the reactive near neighbor strain can be explained through genetic sequence analysis. Importantly, we show the AMD LFI is capable of detecting CPS in a variety of patient samples. The LFI is currently being evaluated in Thailand and Australia; the focus is to optimize and validate testing procedures on melioidosis patient samples prior to initiation of a large, multisite pre-clinical evaluation.

A Switchable Linker-Based Immunoassay for Ultrasensitive Visible Detection of Salmonella in Tomatoes.

Science.gov (United States)

Hahn, Jungwoo; Kim, Eunghee; You, Young Sang; Gunasekaran, Sundaram; Lim, Seokwon; Choi, Young Jin

2017-10-01

On-site detection for sensitive identification of foodborne pathogens on fresh produce with minimal use of specialized instrumentation is crucial to the food industry. A switchable linker (SL)-based immunoassay was designed for ultrasensitive on-site detection of Salmonella in tomato samples. The assay is based on large-scale aggregation of gold nanoparticles (GNPs), induced by a quantitative relationship among the biotinylated Salmonella polyclonal antibody (b-Ab) used as the SL, the functionalized GNPs, and Salmonella. Important factors such as the concentration of SLs, time required for large-scale aggregation, and selectivity of b-Ab were optimized to minimize the detection time (within 45 min with gentle agitation) and achieve the lowest limit of detection (LOD; 10 CFU/g in tomato samples) possible. This SL-based immunoassay with its relatively low LOD and short detection time may meet the need for rapid, simple, on-site analysis of pathogens in fresh produce. The novel switchable linker-based immunoassay is a rapid, specific, and sensitive method that has potential applications for routine diagnostics of Salmonella in tomato products. These advantages make it a practical approach for general use in the processing industry to detect Salmonella rapidly and to implement appropriate regulatory procedures. Furthermore, it could be applied to other fresh products including cantaloupe, strawberry, and cucumbers. © 2017 Institute of Food Technologists®.

Spectro-electrochemical and DFT study of tenoxicam metabolites formed by electrochemical oxidation

International Nuclear Information System (INIS)

Ramírez-Silva, M.T.; Guzmán-Hernández, D.S.; Galano, A.; Rojas-Hernández, A.; Corona-Avendaño, S.; Romero-Romo, M.; Palomar-Pardavé, M.

2013-01-01

Highlights: • Tenoxicam deprotonation and electrochemical oxidation were studied. • Both spectro-electrochemical and theoretical DFT studies were considered. • It was found that the ampholitic species of tenoxicam is a zwitterion. • Electrochemical oxidation of tenoxicam yields two non-electroactive products. • The nature of these fragments was further confirmed by a chromatography study. -- Abstract: From experimental (spectro-electrochemical) and theoretical (DFT) studies, the mechanisms of tenoxicam deprotonation and electrochemical oxidation were assessed. From these studies, new insights on the nature of the ampholitic species involved during tenoxicam's deprotonation in aqueous solution are presented; see scheme A. Moreover, it is shown that, after the analysis of two different reaction schemes that involve up to 10 different molecules and 12 reaction paths, the electrochemical oxidation of tenoxicam, yields two non-electroactive products that are predominately formed by its fragmentation, after the loss of two electrons. The nature of these fragments was further confirmed by a chromatography study

Linkage of biomolecules to solid phases for immunoassay

International Nuclear Information System (INIS)

Chapman, R.S.

1998-01-01

Topics covered by this lecture include a brief review of the principal methods of linkage of biomolecules to solid phase matrices. Copies of the key self explanatory slides are presented as figures together with reprints of two publications by the author dealing with a preferred chemistry for the covalent linkage of antibodies to hydroxyl and amino functional groups and the effects of changes in solid phase matrix and antibody coupling chemistry on the performance of a typical excess reagent immunoassay for thyroid stimulating hormone

Charge Transient, Electrochemical and Impedance Measurements as Tools for Characterization of Nano-Heterostructural Organic/Inorganic Semiconductors

Czech Academy of Sciences Publication Activity Database

Schauer, F.; Nadáždy, V.; Gmucová, K.; Weiss, M.; Kuřitka, I.; Rohovec, Jan; Toušek, J.; Toušková, J.; Lányi, Š.

2013-01-01

Roč. 5, č. 4 (2013), s. 439-443 ISSN 1941-4900 Institutional support: RVO:67985831 Keywords : charge transient spectroscopy * electrochemical spectroscopy * electron structure spectroscopy * organic materials electron structure Subject RIV: BM - Solid Matter Physics ; Magnetism Impact factor: 1.444, year: 2013

Diagnostic effectiveness of immunoassays systems for hepatitis C virus in samples from multi-transfusion patients

International Nuclear Information System (INIS)

Rivero Jimenez, Rene A; Merlin Linares, Julio C; Blanco de Armas, Madelin; Navea Leyva, Leonor M

2009-01-01

Hepatitis C virus (CHV) blood-transmission is a health problem in Cuba and in the world. Some types of diagnostic immunoassays have been developed for the blood certification and in general have a high diagnostic sensitivity and specificity in healthy donors. However, its behavior in samples from multi-transfusion patients could by less effective. To assess the diagnostic effectiveness of the UMELISA HCV third generation Cuban immunoassay (TecnoSUMA, S.A. La Habana), Cuba) in samples from multi-transfusion patients, in parallel, 335 sera from patients were processed by UBI HCV EIA 4.0 (United Biomedical, EE.UU) and UMELISA HCV third generation, and the samples with incongruous results were verified by PCR COBAS AmpliScreen HCV Test, v2 system (Roche, EE.UU.) Comparing the UMELISA HCV third generation system with the UBI HCV EIA 4.0 it was achieved a Sd of 95,8% CI(95%): 92,5-99,15 and a Ed of 100% CI (95%): 99,7-100, with IY: 0,96 (0,93-0,99) with k: 0,0582 ID (95%): 0,9276-0,9888, p = 0,000. Both immunoassay systems were satisfactory for immunodiagnosis of multi-transfusion patients

A review of promising new immunoassay technology for monitoring forest herbicides

Science.gov (United States)

Charles K. McMahon

1993-01-01

Rising costs of classical instrumental methods of chemical analysis coupled with an increasing need for environmental monitoring has lead to the development of highly sensitive, low-cost immunochemical methods of analysis for the detection of environmental contaminants. These methods known simply as immunoassays are chemical assays which use antibodies as reagents. A...

Electrochemical Oxidation of Propene with a LSF15/CGO10 Electrochemical Reactor

DEFF Research Database (Denmark)

Ippolito, Davide; Kammer Hansen, Kent

2014-01-01

A porous electrochemical reactor, made of La0.85Sr0.15FeO3 (LSF) as electrode and Ce0.9Gd0.1O1.95 (CGO) as electrolyte, was studied for the electrochemical oxidation of propene over a wide range of temperatures. Polarization was found to enhance propene oxidation rate. Ce0.9Gd0.1O1.95 was used...... as infiltration material to enhance the effect of polarization on propene oxidation rate, especially at low temperatures. The influence of infiltrated material, as a function of heat treatment, on the reactor electrochemical behavior has been evaluated by using electrochemical impedance spectroscopy...... in suppressing the competing oxygen evolution reaction and promoting the oxidation of propene under polarization, with faradaic efficiencies above 70% at 250◦C. © 2014 The Electrochemical Society....

Trends in interfacial design for surface plasmon resonance based immunoassays

International Nuclear Information System (INIS)

Shankaran, Dhesingh Ravi; Miura, Norio

2007-01-01

Immunosensors based on surface plasmon resonance (SPR) have become a promising tool in sensor technology for biomedical, food, environmental, industrial and homeland security applications. SPR is a surface sensitive optical technique, suitable for real-time and label-free analysis of biorecognition events at functional transducer surfaces. Fabrication of highly active and robust sensing surfaces is an important part in immunoassays because the quality, quantity, chemistry and topography of the interfacial biomembranes play a major role in immunosensor performance. Eventually, a variety of immobilization methods such as physical adsorption, covalent coupling, Langmuir-Blodgett film, polymer thin film, self-assembly, sol-gel, etc, have been introduced over the years for the immobilization of biomolecules (antibody or antigen) on the transducer surfaces. The selection of an immobilization method for an immunoassay is governed by several factors such as nature and stability of the biomolecules, target analyte, application, detection principle, mode of signal transduction, matrix complexity, etc. This paper provides an overview of the various surface modification methods for SPR based immunosensor fabrication. The preparation, structure and application of different functional interfacial surfaces have been discussed along with a brief introduction to the SPR technology, biomolecules and detection principles. (review article)

Trends in interfacial design for surface plasmon resonance based immunoassays

Energy Technology Data Exchange (ETDEWEB)

Shankaran, Dhesingh Ravi [Art, Science and Technology Center for Cooperative Research, Kyushu University, Kasuga-shi, Fukuoka, 816-8580 (Japan); Miura, Norio [Art, Science and Technology Center for Cooperative Research, Kyushu University, Kasuga-shi, Fukuoka, 816-8580 (Japan)

2007-12-07

Immunosensors based on surface plasmon resonance (SPR) have become a promising tool in sensor technology for biomedical, food, environmental, industrial and homeland security applications. SPR is a surface sensitive optical technique, suitable for real-time and label-free analysis of biorecognition events at functional transducer surfaces. Fabrication of highly active and robust sensing surfaces is an important part in immunoassays because the quality, quantity, chemistry and topography of the interfacial biomembranes play a major role in immunosensor performance. Eventually, a variety of immobilization methods such as physical adsorption, covalent coupling, Langmuir-Blodgett film, polymer thin film, self-assembly, sol-gel, etc, have been introduced over the years for the immobilization of biomolecules (antibody or antigen) on the transducer surfaces. The selection of an immobilization method for an immunoassay is governed by several factors such as nature and stability of the biomolecules, target analyte, application, detection principle, mode of signal transduction, matrix complexity, etc. This paper provides an overview of the various surface modification methods for SPR based immunosensor fabrication. The preparation, structure and application of different functional interfacial surfaces have been discussed along with a brief introduction to the SPR technology, biomolecules and detection principles. (review article)

Universal quantum dot-based sandwich-like immunoassay strategy for rapid and ultrasensitive detection of small molecules using portable and reusable optofluidic nano-biosensing platform

International Nuclear Information System (INIS)

Zhou, Liping; Zhu, Anna; Lou, Xuening; Song, Dan; Yang, Rong; Shi, Hanchang; Long, Feng

2016-01-01

A universal sandwich-like immunoassay strategy based on quantum-dots immunoprobe (QD-labeled anti-mouse IgG antibody) was developed for rapid and ultrasensitive detection of small molecules. A portable and reusable optofluidic nano-biosensing platform was applied to investigate the sandwich-like immunoassay mechanism and format of small molecules, as well as the binding kinetics between QD immunoprobe and anti-small molecule antibody. A two-step immunoassay method that involves pre-incubation mixture of different concentration of small molecule and anti-small molecule antibody, and subsequent introduction of QD immunoprobe into the optofluidic cell was conducted for small molecule determination. Compared with the one-step immunoassay method, the two-step immunoassay method can obtain higher fluorescence signal and higher sensitivity index, thus improving the nano-biosensing performance. Based on the proposed strategy, two mode targets, namely, microcystin-LR (MC-LR) and Bisphenol A (BPA) were tested with high sensitivity, rapidity, and ease of use. A higher concentration of small molecules in the sample led to less anti-small molecule antibody bound with antigen-carrier protein conjugate immobilized onto the sensor surface, and less QD immunoprobes bound with anti-small molecule antibody. This phenomenon lowered the fluorescence signal detected by nano-biosensing platform. Under optimal operating conditions, MC-LR and BPA exhibited a limit of detection of 0.003 and 0.04 μg/L, respectively. The LODs were better than those of the indirect competitive immunoassay method for small molecules via Cy5.5-labeled anti-small molecule antibody. The proposed QD-based sandwich-like immunoassay strategy was evaluated in spiked water samples, and showed good recovery, precision and accuracy without complicated sample pretreatments. All these results demonstrate that the new detection strategy could be readily applied to the other trace small molecules in real water samples

Universal quantum dot-based sandwich-like immunoassay strategy for rapid and ultrasensitive detection of small molecules using portable and reusable optofluidic nano-biosensing platform

Energy Technology Data Exchange (ETDEWEB)

Zhou, Liping; Zhu, Anna; Lou, Xuening; Song, Dan; Yang, Rong [School of Environment and Natural Resources, Renmin University of China, Beijing (China); Shi, Hanchang [School of Environment, Tsinghua University, Beijing (China); Long, Feng, E-mail: longf04@ruc.edu.cn [School of Environment and Natural Resources, Renmin University of China, Beijing (China)

2016-01-28

A universal sandwich-like immunoassay strategy based on quantum-dots immunoprobe (QD-labeled anti-mouse IgG antibody) was developed for rapid and ultrasensitive detection of small molecules. A portable and reusable optofluidic nano-biosensing platform was applied to investigate the sandwich-like immunoassay mechanism and format of small molecules, as well as the binding kinetics between QD immunoprobe and anti-small molecule antibody. A two-step immunoassay method that involves pre-incubation mixture of different concentration of small molecule and anti-small molecule antibody, and subsequent introduction of QD immunoprobe into the optofluidic cell was conducted for small molecule determination. Compared with the one-step immunoassay method, the two-step immunoassay method can obtain higher fluorescence signal and higher sensitivity index, thus improving the nano-biosensing performance. Based on the proposed strategy, two mode targets, namely, microcystin-LR (MC-LR) and Bisphenol A (BPA) were tested with high sensitivity, rapidity, and ease of use. A higher concentration of small molecules in the sample led to less anti-small molecule antibody bound with antigen-carrier protein conjugate immobilized onto the sensor surface, and less QD immunoprobes bound with anti-small molecule antibody. This phenomenon lowered the fluorescence signal detected by nano-biosensing platform. Under optimal operating conditions, MC-LR and BPA exhibited a limit of detection of 0.003 and 0.04 μg/L, respectively. The LODs were better than those of the indirect competitive immunoassay method for small molecules via Cy5.5-labeled anti-small molecule antibody. The proposed QD-based sandwich-like immunoassay strategy was evaluated in spiked water samples, and showed good recovery, precision and accuracy without complicated sample pretreatments. All these results demonstrate that the new detection strategy could be readily applied to the other trace small molecules in real water samples

False-positive ethyl glucuronide immunoassay screening caused by a propyl alcohol-based hand sanitizer.

Science.gov (United States)

Arndt, Torsten; Grüner, Joachim; Schröfel, Stefanie; Stemmerich, Karsten

2012-11-30

Urine ethyl glucuronide (EtG) is considered as a specific marker of recent ethanol consumption. We describe false-positive DRI(®) EIA EtG enzyme immunoassay results caused by propyl glucuronides in urine after using a propanol-based hand sanitizer. EtG screening was done with the DRI(®) EIA EtG assay (Microgenics), using a cut-off of 0.5 mg/L as recommended by the manufacturer and of 0.1 mg/L as demanded by the German Regulations for Reissuing Drivers Licenses. Confirmatory EtG analysis was done with the ClinMass(®) EtG LC-MS/MS testkit (Recipe), extended by the mass transitions 235.1→75.1, 235.1→85.1, and 235.1→113.1 for the detection of the 1- and 2-propyl glucuronides. Self-experiments were done by staff members of our lab (n=7), using 3 mL Sterillium(®) Classic Pure (30 g/100 g 1-propanol and 45 g/100 g 2-propanol) for hand sanitation every quarter of an hour for 8 h according to DIN EN 1500:2011-05 with and without an exhauster and by passive inhalation of the sanitizer vapor. Spot urine samples were taken immediately before and up to 24 h after the first sanitizer use. False-positive immunoassay results of up to 4 mg/L or 2.3 mg/g creatinine were obtained after normal use of the sanitizer and also after passive inhalation of the sanitizer vapor (up to 0.89 mg/L or 0.61 mg/g). Immunoassay results were positive even after 4-fold use of the sanitizer (up to 0.14 mg/L or 0.38 mg/g) and up to 6 h after the last sanitizer contact (maximum 0.63 mg/L and 0.33 mg/g for sanitizer users and 0.25 mg/g after passive inhalation). Spiking of EtG-free urine with 1-propyl glucuronide (Athena Environmental Sciences) between 0.05 and 10 mg/L clearly demonstrated a cross reaction of the immunoassay of approx. 10% as compared to EtG. LC-MS/MS of urines with a positive immunoassay EtG result did not show EtG signals, but distinct signals of 1-propyl glucuronide (n-propyl glucuronide) and 2-propyl glucuronide (iso-propyl glucuronide). An exhauster effectively prevented

Electrochemical and AFM Characterization of G-Quadruplex Electrochemical Biosensors and Applications

Science.gov (United States)

2018-01-01

Guanine-rich DNA sequences are able to form G-quadruplexes, being involved in important biological processes and representing smart self-assembling nanomaterials that are increasingly used in DNA nanotechnology and biosensor technology. G-quadruplex electrochemical biosensors have received particular attention, since the electrochemical response is particularly sensitive to the DNA structural changes from single-stranded, double-stranded, or hairpin into a G-quadruplex configuration. Furthermore, the development of an increased number of G-quadruplex aptamers that combine the G-quadruplex stiffness and self-assembling versatility with the aptamer high specificity of binding to a variety of molecular targets allowed the construction of biosensors with increased selectivity and sensitivity. This review discusses the recent advances on the electrochemical characterization, design, and applications of G-quadruplex electrochemical biosensors in the evaluation of metal ions, G-quadruplex ligands, and other small organic molecules, proteins, and cells. The electrochemical and atomic force microscopy characterization of G-quadruplexes is presented. The incubation time and cations concentration dependence in controlling the G-quadruplex folding, stability, and nanostructures formation at carbon electrodes are discussed. Different G-quadruplex electrochemical biosensors design strategies, based on the DNA folding into a G-quadruplex, the use of G-quadruplex aptamers, or the use of hemin/G-quadruplex DNAzymes, are revisited. PMID:29666699

Magnetic filtration of phase separating ferrofluids: From basic concepts to microfluidic device

Science.gov (United States)

Kuzhir, P.; Magnet, C.; Ezzaier, H.; Zubarev, A.; Bossis, G.

2017-06-01

In this work, we briefly review magnetic separation of ferrofluids composed of large magnetic particles (60 nm of the average size) possessing an induced dipole moment. Such ferrofluids exhibit field-induced phase separation at relatively low particle concentrations (∼0.8 vol%) and magnetic fields (∼10 kA/m). Particle aggregates appearing during the phase separation are extracted from the suspending fluid by magnetic field gradients much easier than individual nanoparticles in the absence of phase separation. Nanoparticle capture by a single magnetized microbead and by multi-collector systems (packed bed of spheres and micro-pillar array) has been studied both experimentally and theoretically. Under flow and magnetic fields, the particle capture efficiency Λ decreases with an increasing Mason number for all considered geometries. This decrease may become stronger for aggregated magnetic particles (Λ ∝Ma-1.7) than for individual ones (Λ ∝Ma-1) if the shear fields are strong enough to provoke aggregate rupture. These results can be useful for development of new magneto-microfluidic immunoassays based on magnetic nanoparticles offering a much better sensitivity as compared to presently used magnetic microbeads.

Comparison of Immunoassay methods for T3, T4 and TSH

International Nuclear Information System (INIS)

Alonso Rodríguez, Celia A.

2016-01-01

Measurements of T3, T4 and TSH have been considered very important in the diagnosis and monitoring of thyroid diseases both overt and subclinical. These subclinical diseases are actively seeking for years, both in healthy patients and hospitalized for other illnesses; and in the population over 35 years, especially women, in health checkups. The active search for these diseases requires the use of rapid and reliable techniques; that can be developed massively, with good level of detectability and comparable. The overall objective is to present the evaluation of different immunoassay techniques with respect to the RIA and IRMA: ELISA, chemiluminescence, Amplified Chemiluminescence, electrochemiluminescence Immunofluorescence. Compare including automatic methods and analyze the cost and feasibility of them for laboratory immunoassay. ELISA colorimetric technique for dosing was comparable to RIA T4, not for T3. Chemiluminescence (AMERLITE) compared to dosing RIA and IRMA T4 to TSH proved to be valid for both. Amplified Chemiluminescence (Immulite) compared to IRMA for TSH was no significant difference. Electrochemiluminescence (Elecsys 2010) compared to T3 and T4 RIA and IRMA for TSH, no significant differences for T4 and TSH; but no variation to T3. Immunofluorescence (AIA-600) used to compare with RIA for T3 and T4, and TSH IRMA, no significant differences for the measured analytes. Benchmarking of automatic methods suggests that the most thrifty of trials is Immunofluorescence the AIA-600, regarding calibration and control, programming time, randomization and the ability to save the value of the fluorescence deferred calculations for tests without valid at the time of realizing calibration. Analyzing the cost and feasibility of these methods for laboratory immunoassay, we must consider that their characteristics electrochemiluminescence is the fastest, but its price is prohibitive for our health systems. The AIA-600 appears to be the method of choice for its

Development and analytical performance of a new ARCHITECT automated dipeptidyl peptidase-4 immunoassay

Directory of Open Access Journals (Sweden)

Philip M. Hemken

2017-12-01

Full Text Available Background: Dipeptidyl peptidase-4 (DPP-4 may be a suitable biomarker to identify people with severe asthma who have greater activation of the interleukin-13 (IL-13 pathway and may therefore benefit from IL-13-targeted treatments. We report the analytical performance of an Investigational Use Only immunoassay and provide data on the biological range of DPP-4 concentrations. Methods: We assessed assay performance, utilising analyses of precision, linearity and sensitivity; interference from common endogenous assay interferents, and from asthma and anti-diabetic medications, were also assessed. The assay was used to measure the range of serum DPP-4 concentrations in healthy volunteers and subjects with diabetes and severe, uncontrolled asthma. Results: The total precision of DPP-4 concentration measurement (determined using percentage coefficient of variation was ≤5% over 20 days. Dilution analysis yielded linear results from 30 to 1305 ng/mL; the limit of quantitation was 19.2 ng/mL. No notable endogenous or drug interferences were observed at the expected therapeutic concentration. Median DPP-4 concentrations in healthy volunteers and subjects with asthma or Type 1 diabetes were assessed, with concentrations remaining similar in subjects with diabetes and asthma across different demographics. Conclusion: These analyses indicate that the ARCHITECT DPP-4 Immunoassay is a reliable and robust method for measuring serum DPP-4 concentration. Keywords: Asthma, Automated immunoassay, Biomarker, Dipeptidyl peptidase-4, IL-13

Direct biosensor immunoassays for the detection of nonmilk proteins in milk powder

NARCIS (Netherlands)

Haasnoot, W.; Olieman, K.; Cazemier, G.; Verheijen, R.

2001-01-01

The low prices of some nonmilk proteins make them attractive as potential adulterants in dairy products. An optical biosensor (BIACORE 3000) was used to develop a direct and combined biosensor immunoassay (BIA) for the simultaneous detection of soy, pea, and soluble wheat proteins in milk powders.

A competitive chemiluminescence enzyme immunoassay for rapid and sensitive determination of enrofloxacin

Science.gov (United States)

Yu, Fei; Wu, Yongjun; Yu, Songcheng; Zhang, Huili; Zhang, Hongquan; Qu, Lingbo; Harrington, Peter de B.

With alkaline phosphatase (ALP)-adamantane (AMPPD) system as the chemiluminescence (CL) detection system, a highly sensitive, specific and simple competitive chemiluminescence enzyme immunoassay (CLEIA) was developed for the measurement of enrofloxacin (ENR). The physicochemical parameters, such as the chemiluminescent assay mediums, the dilution buffer of ENR-McAb, the volume of dilution buffer, the monoclonal antibody concentration, the incubation time, and other relevant variables of the immunoassay have been optimized. Under the optimal conditions, the detection linear range of 350-1000 pg/mL and the detection limit of 0.24 ng/mL were provided by the proposed method. The relative standard deviations were less than 15% for both intra and inter-assay precision. This method has been successfully applied to determine ENR in spiked samples with the recovery of 103%-96%. It showed that CLEIA was a good potential method in the analysis of residues of veterinary drugs after treatment of related diseases.

Screening for cocaine on Euro banknotes by a highly sensitive enzyme immunoassay.

Science.gov (United States)

Abdelshafi, Nahla A; Panne, Ulrich; Schneider, Rudolf J

2017-04-01

This study focused on quantitative detection of cocaine on Euro banknotes in Germany. A sensitive direct competitive immunoassay was developed and optimized with a limit of detection (LOD) of 5.6ng/L. Exhaustive cocaine extraction by solvent was tested using different methanol concentrations and buffered solutions. Cross-reactivity studies were performed to determine the degree of interference of cocaine metabolites with the immunoassay. Sixty-five Euro banknotes obtained from different districts in Berlin were evaluated. A 100% contamination frequency with cocaine was detected. A comparison between the amount of cocaine extracted by cotton swabbing of one square centimeter of the banknote showed a good correlation for lower contamination levels. This assay showed high sensitivity of detecting pg of cocaine per 1cm 2 of one banknote by swabbing 1cm 2 : 0, 14, and 21pg/cm 2 . Moreover, three notes of different denominations revealed high cocaine concentration; 1.1mg/note, and twice 55µg/note. Copyright © 2017 Elsevier B.V. All rights reserved.

Tuberculin purified protein derivative (PPD) immunoassay as an in vitro alternative assay for identity and confirmation of potency.

Science.gov (United States)

Ho, Mei M; Kairo, Satnam K; Corbel, Michael J

2006-01-01

Tuberculin purified protein derivative (PPD) currently can only be standardised by delayed hypersensitivity skin reactions in sensitised guinea pigs. An in vitro dot blot immunoassay was developed for both identity and confirmation of potency estimation of PPD. Polyclonal antibodies (mainly IgG) were generated and immunoreacted with human, bovine and, to lesser extent, avian PPD preparations. Combining size exclusion chromatography (FPLC-SEC) and dot blot immunoassay, the results showed that PPD preparations were mixtures of very heterogeneous tuberculoproteins ranging in size from very large aggregates to very small degraded molecules. All individual fractions of PPD separated by size were immunoreactive, although those of the largest molecular sizes appeared the most immunoreactive in this in vitro dot blot immunoassay. This method is very sensitive and specific to tuberculoproteins and can be an in vitro alternative for the in vivo intradermal skin assay which uses guinea pigs for identity of PPD preparations. Although the capacity of PPD to elicit cell-mediated immune responses on intradermal testing has to be confirmed by in vivo assay, the dot blot immunoassay offers a rapid, sensitive and animal-free alternative to in vivo testing for confirming the identity of PPD preparations with appropriate potencies. This alternative assay would be particularly useful for national regulatory laboratories for confirming the data of manufacturers and thus reducing the use of animals.

Development of Indirect Competitive Immuno-Assay Method Using SPR Detection for Rapid and Highly Sensitive Measurement of Salivary Cortisol Levels

International Nuclear Information System (INIS)

Tahara, Yusuke; Huang, Zhe; Kiritoshi, Tetsuro; Onodera, Takeshi; Toko, Kiyoshi

2014-01-01

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 immuno-assay method based on an indirect competitive method using a commercially available surface plasmon resonance instrument. The surface of an Au chip was modified with PEG6-COOH aromatic dialkanethiol self-assembled monolayers and hydrocortisone 3-(O-carboxymethyl) oxime (hydrocortisone 3-CMO) as a cortisol analog. A detection limit of 38 ppt range with a measurement range of 10 ppt–100 ppb was accomplished without the incubation of a mixing solution consisting of standard cortisol and an anti-cortisol antibody, and the time for quantification of cortisol concentration was 8 min from the sample injection. We experimentally compared our immuno-assay 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 immuno-assay had a good correlation with those obtained by ELISA assay (R = 0.96). Our findings indicate the potential utility of the cortisol immuno-assay for measurements of human salivary cortisol levels.

Development of Indirect Competitive Immuno-Assay Method Using SPR Detection for Rapid and Highly Sensitive Measurement of Salivary Cortisol Levels

Energy Technology Data Exchange (ETDEWEB)

Tahara, Yusuke; Huang, Zhe; Kiritoshi, Tetsuro [Graduate School of Information Science and Electrical Engineering, Kyushu University, Fukuoka (Japan); Onodera, Takeshi [Research and Development Center for Taste and Odor Sensing, Kyushu University, Fukuoka (Japan); Toko, Kiyoshi, E-mail: toko@ed.kyushu-u.ac.jp [Graduate School of Information Science and Electrical Engineering, Kyushu University, Fukuoka (Japan); Research and Development Center for Taste and Odor Sensing, Kyushu University, Fukuoka (Japan)

2014-05-30

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 immuno-assay method based on an indirect competitive method using a commercially available surface plasmon resonance instrument. The surface of an Au chip was modified with PEG6-COOH aromatic dialkanethiol self-assembled monolayers and hydrocortisone 3-(O-carboxymethyl) oxime (hydrocortisone 3-CMO) as a cortisol analog. A detection limit of 38 ppt range with a measurement range of 10 ppt–100 ppb was accomplished without the incubation of a mixing solution consisting of standard cortisol and an anti-cortisol antibody, and the time for quantification of cortisol concentration was 8 min from the sample injection. We experimentally compared our immuno-assay 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 immuno-assay had a good correlation with those obtained by ELISA assay (R = 0.96). Our findings indicate the potential utility of the cortisol immuno-assay for measurements of human salivary cortisol levels.

Electrochemical thermodynamic measurement system

Science.gov (United States)

Reynier, Yvan [Meylan, FR; Yazami, Rachid [Los Angeles, CA; Fultz, Brent T [Pasadena, CA

2009-09-29

The present invention provides systems and methods for accurately characterizing thermodynamic and materials properties of electrodes and electrochemical energy storage and conversion systems. Systems and methods of the present invention are configured for simultaneously collecting a suite of measurements characterizing a plurality of interconnected electrochemical and thermodynamic parameters relating to the electrode reaction state of advancement, voltage and temperature. Enhanced sensitivity provided by the present methods and systems combined with measurement conditions that reflect thermodynamically stabilized electrode conditions allow very accurate measurement of thermodynamic parameters, including state functions such as the Gibbs free energy, enthalpy and entropy of electrode/electrochemical cell reactions, that enable prediction of important performance attributes of electrode materials and electrochemical systems, such as the energy, power density, current rate and the cycle life of an electrochemical cell.

One step paired electrochemical synthesis of iron and iron oxide nanoparticles

Directory of Open Access Journals (Sweden)

Ordoukhanian Juliet

2016-09-01

Full Text Available In this study, a new one step paired electrochemical method is developed for simultaneous synthesis of iron and iron oxide nanoparticles. iron and iron oxide are prepared as cathodic and anodic products from iron (ii sulfate aqueous solution in a membrane divided electrolytic cell by the pulsed current electrosynthesis. Because of organic solvent-free and electrochemical nature of the synthesis, the process could be considered as green and environmentally friendly. The reduction of energy consumption and low cost are the other significant advantages of this new method that would have a great application potential in the chemical industry. The nanostructure of prepared samples was characterized by Fourier transform infrared spectroscopy (FT-IR, X-ray diffraction (XRD, scanning electron microscopy (SEM and transmission electron microscopy (TEM. The magnetic properties were studied by vibrating sample magnetometer (VsM.

Single biosensor immunoassay for the detection of five aminoglycosides in reconstituted skimmed milk

NARCIS (Netherlands)

Haasnoot, W.; Cazemier, G.; Koets, M.; Amerongen, van A.

2003-01-01

The application of an optical biosensor (Biacore 3000), with four flow channels (Fcs), in combination with a mixture of four specific antibodies resulted in a competitive inhibition biosensor immunoassay (BIA) for the simultaneous detection of the five relevant aminoglycosides in reconstituted

Magnetic immunoassay using CdSe/ZnS quantum dots as fluorescent probes to detect the level of DNA methyltransferase 1 in human serum sample

Directory of Open Access Journals (Sweden)

Yu F

2018-01-01

than ELISA kits.Conclusion: These indicated that the proposed FLISA method was sensitive and high throughput and can quickly screen the level of DNMT1 in serum samples. Keywords: DNA methyltransferase 1, quantum dots, fluorescence immunoassay, magnetic carboxyl beads, high throughput, serum sample

Chemiluminescence immunoassay for prostate-specific antigen

International Nuclear Information System (INIS)

Zhang Xuefeng; Liu Yibing; Jia Juanjuan; Xu Wenge; Li Ziying; Chen Yongli; Han Shiquan

2008-01-01

The chemiluminescence immunoassay (CLIA) for serum total prostate-specific antigen (T-PSA) was developed. The reaction of luminol with hydrogen peroxide was introduced into this chemiluminescence system. The detection limit is established as 0.12 μg/L (n=10, mean of zero standard + 2SD) and the analytical recovery of PSA is 83.8%-118.7%. The intra-assay and inter-assay CVs vary from 4.4%-5.0% and 6.2%-11.7%, respectively. The experimental correlation coefficient of dilution is found to be 0.999. Compared with immunoradiometric assay (IRMA) kits, the correlative equation is y=1.07x+0.68, and correlation coefficient r=0.97. The standard range for the method is 1.5-80 μg/L, and it presents good linearity. (authors)

Inkjet-printed point-of-care immunoassay on a nanoscale polymer brush enables subpicomolar detection of analytes in blood

Science.gov (United States)

Joh, Daniel Y.; Hucknall, Angus M.; Wei, Qingshan; Mason, Kelly A.; Lund, Margaret L.; Fontes, Cassio M.; Hill, Ryan T.; Blair, Rebecca; Zimmers, Zackary; Achar, Rohan K.; Tseng, Derek; Gordan, Raluca; Freemark, Michael; Ozcan, Aydogan; Chilkoti, Ashutosh

2017-08-01

The ELISA is the mainstay for sensitive and quantitative detection of protein analytes. Despite its utility, ELISA is time-consuming, resource-intensive, and infrastructure-dependent, limiting its availability in resource-limited regions. Here, we describe a self-contained immunoassay platform (the “D4 assay”) that converts the sandwich immunoassay into a point-of-care test (POCT). The D4 assay is fabricated by inkjet printing assay reagents as microarrays on nanoscale polymer brushes on glass chips, so that all reagents are “on-chip,” and these chips show durable storage stability without cold storage. The D4 assay can interrogate multiple analytes from a drop of blood, is compatible with a smartphone detector, and displays analytical figures of merit that are comparable to standard laboratory-based ELISA in whole blood. These attributes of the D4 POCT have the potential to democratize access to high-performance immunoassays in resource-limited settings without sacrificing their performance.

Magnetoresistive biosensors for quantitative proteomics

Science.gov (United States)

Zhou, Xiahan; Huang, Chih-Cheng; Hall, Drew A.

2017-08-01

Quantitative proteomics, as a developing method for study of proteins and identification of diseases, reveals more comprehensive and accurate information of an organism than traditional genomics. A variety of platforms, such as mass spectrometry, optical sensors, electrochemical sensors, magnetic sensors, etc., have been developed for detecting proteins quantitatively. The sandwich immunoassay is widely used as a labeled detection method due to its high specificity and flexibility allowing multiple different types of labels. While optical sensors use enzyme and fluorophore labels to detect proteins with high sensitivity, they often suffer from high background signal and challenges in miniaturization. Magnetic biosensors, including nuclear magnetic resonance sensors, oscillator-based sensors, Hall-effect sensors, and magnetoresistive sensors, use the specific binding events between magnetic nanoparticles (MNPs) and target proteins to measure the analyte concentration. Compared with other biosensing techniques, magnetic sensors take advantage of the intrinsic lack of magnetic signatures in biological samples to achieve high sensitivity and high specificity, and are compatible with semiconductor-based fabrication process to have low-cost and small-size for point-of-care (POC) applications. Although still in the development stage, magnetic biosensing is a promising technique for in-home testing and portable disease monitoring.

On-chip determination of C-reactive protein using magnetic particles in continuous flow.

Science.gov (United States)

Phurimsak, Chayakom; Tarn, Mark D; Peyman, Sally A; Greenman, John; Pamme, Nicole

2014-11-04

We demonstrate the application of a multilaminar flow platform, in which functionalized magnetic particles are deflected through alternating laminar flow streams of reagents and washing solutions via an external magnet, for the rapid detection of the inflammatory biomarker, C-reactive protein (CRP). The two-step sandwich immunoassay was accomplished in less than 60 s, a vast improvement on the 80-300 min time frame required for enzyme-linked immunosorbent assays (ELISA) and the 50 min necessary for off-chip magnetic particle-based assays. The combination of continuous flow and a stationary magnet enables a degree of autonomy in the system, while a detection limit of 0.87 μg mL(-1) makes it suitable for the determination of CRP concentrations in clinical diagnostics. Its applicability was further proven by assaying real human serum samples and comparing those results to values obtained using standard ELISA tests.

Magnetic nanowires (Fe, Fe-Co, Fe-Ni – magnetic moment reorientation in respect of wires composition

Directory of Open Access Journals (Sweden)

Kalska-Szostko Beata

2015-03-01

Full Text Available Magnetic nanowires of Fe, Fe-Co, and Fe-Ni alloy and layered structure were prepared by electrochemical alternating current (AC deposition method. The morphology of the nanowires in and without the matrix was studied by energy dispersive X-ray spectroscopy (EDX, scanning electron microscopy (SEM, and X-ray diffraction (XRD, respectively. The wires either show strong dependence on the combination of elements deposition (alloy or layered or chemical composition (Co or Ni. The magnetic properties of the nanostructures were determined on the basis of Mössbauer spectroscopy (MS.

A novel in situ electrochemical NMR cell with a palisade gold film electrode

Science.gov (United States)

Ni, Zu-Rong; Cui, Xiao-Hong; Cao, Shuo-Hui; Chen, Zhong

2017-08-01

In situ electrochemical nuclear magnetic resonance (EC-NMR) has attracted considerable attention because of its ability to directly observe real-time electrochemical processes. Therefore, minimizing the incompatibility between the electrochemical device and NMR detection has become an important challenge. A circular thin metal film deposited on the outer surface of a glass tube with a thickness considerably less than the metal skin depth is considered to be the ideal working electrode. In this study, we demonstrate that such a thin film electrode still has a great influence on the radio frequency field homogeneity in the detective zone of the NMR spectrometer probe and provide theoretical and experimental confirmation of its electromagnetic shielding. Furthermore, we propose a novel palisade gold film device to act as the working electrode. The NMR nutation behavior of protons shows that the uniformity of the radio frequency field is greatly improved, increasing the sensitivity in NMR detection. Another advantage of the proposed device is that an external reference standard adapted to the reaction compound can be inserted as a probe to determine the fluctuation of the physico-chemical environment and achieve high-accuracy quantitative NMR analysis. A three-chamber electrochemical device based on the palisade gold film design was successfully fabricated and the in situ electrochemical NMR performance was validated in a standard 5 mm NMR probe by acquiring voltammograms and high-resolution NMR spectra to characterize the electrochemically generated species. The evolution of in situ EC-NMR spectrum monitoring of the redox transformation between p-benzoquinone and hydroquinone demonstrates the ability of the EC-NMR device to simultaneously quantitatively determine the reactants and elucidate the reaction mechanism at the molecular level.

A novel in situ electrochemical NMR cell with a palisade gold film electrode

Directory of Open Access Journals (Sweden)

Zu-Rong Ni

2017-08-01

Full Text Available In situ electrochemical nuclear magnetic resonance (EC-NMR has attracted considerable attention because of its ability to directly observe real-time electrochemical processes. Therefore, minimizing the incompatibility between the electrochemical device and NMR detection has become an important challenge. A circular thin metal film deposited on the outer surface of a glass tube with a thickness considerably less than the metal skin depth is considered to be the ideal working electrode. In this study, we demonstrate that such a thin film electrode still has a great influence on the radio frequency field homogeneity in the detective zone of the NMR spectrometer probe and provide theoretical and experimental confirmation of its electromagnetic shielding. Furthermore, we propose a novel palisade gold film device to act as the working electrode. The NMR nutation behavior of protons shows that the uniformity of the radio frequency field is greatly improved, increasing the sensitivity in NMR detection. Another advantage of the proposed device is that an external reference standard adapted to the reaction compound can be inserted as a probe to determine the fluctuation of the physico-chemical environment and achieve high-accuracy quantitative NMR analysis. A three-chamber electrochemical device based on the palisade gold film design was successfully fabricated and the in situ electrochemical NMR performance was validated in a standard 5 mm NMR probe by acquiring voltammograms and high-resolution NMR spectra to characterize the electrochemically generated species. The evolution of in situ EC-NMR spectrum monitoring of the redox transformation between p-benzoquinone and hydroquinone demonstrates the ability of the EC-NMR device to simultaneously quantitatively determine the reactants and elucidate the reaction mechanism at the molecular level.

Evaluation of field test kits including immunoassays for the detection of contaminants in soil and water

International Nuclear Information System (INIS)

Waters, L.C.; Smith, R.R.; Counts, R.W.; Stewart, J.H.; Jenkins, R.A.

1993-01-01

Effective field test methods are needed for hazardous waste site characterization and remediation. Useful field methods should be rapid, analyte-specific, cost-effective and accurate in the concentration range at which the analyte is regulated. In this study, field test kits for polychlorinated biphenyls (PCBs), mercury, lead and nitrate were evaluated with reference to these criteria. PCBs and mercury, in soils, were analyzed by immunoassay. Ionic lead and nitrate, in water, were measured chemically using test strips. Except for lead, each analyte was measured in both spiked and actual field samples. Twenty to 40 samples per day can be analyzed with the immunoassays and even more with the strip tests. The sensitivity of the immunoassays is in the 1-3 ppM range. Nitrate was consistently detected at ≥5 ppM; lead ions at ≥20 ppM. Results obtained using these methods compared favorably with those obtained by standard laboratory methods. In addition to being useful field screening methods, these kits can be used in the laboratory to sort out negative samples and/or to define proper dilutions for positive samples requiring further analysis

Homogeneous immunoassay for human IgG using oriented hen egg IgY immobilized on gold sol nanoparticles

International Nuclear Information System (INIS)

Yeritsyan, H.E.; Gasparyan, V.K.

2012-01-01

Homogeneous immunoassays using (red) gold nanoparticles represent an attractive detection scheme because of the option of photometric readout. We have applied oriented immobilization of hen egg immunoglobulin Y (IgY) on gold nanoparticles when developing a homogeneous immunoassay for human IgG. In oriented immobilization, as opposed to random immobilization, the antigen binding capabilities of the antibodies are retained. It is shown that such immunoassay has significantly better sensitivity in comparison with methods based on conventional immobilization of affinity-purified antibodies. It is also shown that hen egg IgY is better suited than rabbit antibodies, because much more antibody can be immobilized on gold nanoparticles without any destabilization, probably because of the more acidic nature of these antibodies. In addition, hen egg IgY can be supplied in higher quantity and can be prepared more easily than IgG from rabbits. Bleeding and slaughtering of animals is not needed. The assay presented here has a wide detection range (30-500 ng. mL -1 ) and a limit of detection as low as 30 ng. mL -1 of human IgG. (author)

Electrochemical genoassays on gold-coated magnetic nanoparticles to quantify genetically modified organisms (GMOs) in food and feed as GMO percentage.

Science.gov (United States)

Plácido, Alexandra; Pereira, Clara; Guedes, Alexandra; Barroso, M Fátima; Miranda-Castro, Rebeca; de-Los-Santos-Álvarez, Noemí; Delerue-Matos, Cristina

2018-07-01

The integration of nanomaterials in the field of (bio)sensors has allowed developing strategies with improved analytical performance. In this work, ultrasmall core-shell Fe 3 O 4 @Au magnetic nanoparticles (MNPs) were used as the platform for the immobilization of event-specific Roundup Ready (RR) soybean and taxon-specific DNA sequences. Firstly, monodisperse Fe 3 O 4 MNPs were synthesized by thermal decomposition and subsequently coated with a gold shell through reduction of Au(III) precursor on the surface of the MNPs in the presence of an organic capping agent. This nanosupport exhibited high colloidal stability, average particle size of 10.2 ± 1.3 nm, and spherical shape. The covalent immobilization of ssDNA probe onto the Au shell of the Fe 3 O 4 @Au MNPs was achieved through a self-assembled monolayer (SAM) created from mixtures of alkane thiols (6-mercapto-1-hexanol and mercaptohexanoic acid). The influence of the thiols ratio on the electrochemical performance of the resulting electrochemical genoassays was studied, and remarkably, the best analytical performance was achieved for a pure mercaptohexanoic acid SAM. Two quantification assays were designed; one targeting an RR sequence and a second targeting a reference soybean gene, both with a sandwich format for hybridization, signaling probes labelled with fluorescein isothiocyanate (FITC), enzymatic amplification and chronoamperometric detection at screen-printed carbon electrodes (SPCE). The magnetogenoassays exhibited linear ranges from 0.1 to 10.0 nM and from 0.1 to 5.0 nM with similar detection limits of 0.02 nM and 0.05 nM for the event-specific (RR) and the taxon-specific (lectin) targets, respectively. The usefulness of the approach was demonstrated by its application to detect genetically modified organisms (GMOs) in feed and food. Copyright © 2018 Elsevier B.V. All rights reserved.

Cation-exchange antibody labeling for simultaneous electrochemical detection of tumor markers CA15-3 and CA19-9

International Nuclear Information System (INIS)

Wang, Guangjie; Qing, Yi; Shan, Jinlu; Jin, Feng; Wang, Dong; Yuan, Ruo

2013-01-01

We report on a new kind of non-covalent multi-label electrochemical immunoassay that was applied to simultaneously quantify the tumor markers CA15-3 and CA19-9. The method employs a nanohybrid composed of an ionomer and conductive titanium dioxide nanoparticles that act as a matrix support for the antibodies. The two antibodies (anti-CA153 and anti-CA199) were labeled (a) with a cobaltous dipyridine complex, and (b) with methylene blue. Labeling is based on cation-exchange interaction rather than on covalent conjugation. The redox potentials of the two labels are separated by an interval of 0.3 V. The resulting sandwich-type immunosensor was read out by differential pulse voltammetry. The potential sites and currents of the two redox probes reflect the concentration of the two analytes. The two analytes were determined with a detection limit of 1.6 U mL −1 for CA19-9, and of 0.3 U mL −1 for CA15-3 (author)

Electrochemical analysis

International Nuclear Information System (INIS)

Hwang, Hun

2007-02-01

This book explains potentiometry, voltametry, amperometry and basic conception of conductometry with eleven chapters. It gives the specific descriptions on electrochemical cell and its mode, basic conception of electrochemical analysis on oxidation-reduction reaction, standard electrode potential, formal potential, faradaic current and faradaic process, mass transfer and overvoltage, potentiometry and indirect potentiometry, polarography with TAST, normal pulse and deferential pulse, voltammetry, conductometry and conductometric titration.

A dual amplified electrochemical immunosensor for ofloxacin: Polypyrrole film-Au nanocluster as the matrix and multi-enzyme-antibody functionalized gold nanorod as the label

International Nuclear Information System (INIS)

Zang, Shuai; Liu, Yingju; Lin, Mouhong; Kang, Jianli; Sun, Yuanming; Lei, Hongtao

2013-01-01

Graphical abstract: Schematic representation of the OFL electrochemical immunosensor using Au nanoclusters/PPy/GCE as the substrate and multi-HRP-GNR-Ab2 bioconjugates as the label. Highlights: ► Gold nanorod was used to load HRP and Ab 2 to form multi-HRP-GNR-Ab 2 . ► A sensitive immunosensor for ofloxacin was constructed using the homemade antibody. ► A dual signal amplified strategy was based on the PPy-Au and multi-HRP-GNR-Ab 2 . -- Abstract: In this work, an electrochemical immunosensor, basing on a dual signal amplified strategy by employing a biocompatible polypyrrole film-Au nanocluster matrix as a sensor platform and multi-enzyme-antibody functionalized gold nanorod as an electrochemical detection label, is established for sensitive detection of ofloxacin (OFL). Firstly, polypyrrole film and Au nanoclusters were progressively fabricated onto the surface of a glassy carbon electrode via electropolymerization and electrochemical deposition, respectively. Such PPy-Au nanocomposite modified electrode was used to immobilize OFL-OVA, blocked with the blocking reagent, and then associated with the corresponding antibody. Secondly, gold nanorod (GNR) was synthesized to load horseradish peroxidase (HRP) and horseradish peroxidase-secondary antibody (HRP-Ab 2 ), and the resulting nanostructure (multi-HRP-GNR-Ab 2 ) was applied as the detection label. The fabrication process of the ordered multilayer structure and immunosensor were characterized by scanning electron microscopy (SEM) and electrochemical measurements, respectively. Finally, based on a competitive immunoassay, i.e., the association ability with the corresponding antibody between the captured antigen and free OFL in the solution, the fabricated immunosensor exhibited a sensitive response to OFL in the range from 0.08 to 410 ng/mL with a detection limit of 0.03 ng/mL. The current immunosensor exhibited good sensitivity, selectivity and long-term stability. This amplification strategy shows excellent

Rapid Salmonella detection in experimentally inoculated equine faecal and veterinary hospital environmental samples using commercially available lateral flow immunoassays.

Science.gov (United States)

Burgess, B A; Noyes, N R; Bolte, D S; Hyatt, D R; van Metre, D C; Morley, P S

2015-01-01

Salmonella enterica is the most commonly reported cause of outbreaks of nosocomial infections in large animal veterinary teaching hospitals and the closure of equine hospitals. Rapid detection may facilitate effective control practices in equine populations. Shipping and laboratory testing typically require ≥48 h to obtain results. Lateral flow immunoassays developed for use in food-safety microbiology provide an alternative that has not been evaluated for use with faeces or environmental samples. We aimed to identify enrichment methods that would allow commercially available rapid Salmonella detection systems (lateral flow immunoassays) to be used in clinical practice with equine faecal and environmental samples, providing test results in 18-24 h. In vitro experiment. Equine faecal and environmental samples were inoculated with known quantities of S. enterica serotype Typhimurium and cultured using 2 different enrichment techniques for faeces and 4 enrichment techniques for environmental samples. Samples were tested blindly using 2 different lateral flow immunoassays and plated on agar media for confirmatory testing. In general, commercial lateral flow immunoassays resulted in fewer false-negative test results with enrichment of 1 g faecal samples in tetrathionate for 18 h, while all environmental sample enrichment techniques resulted in similar detection rates. The limit of detection from spiked samples, ∼4 colony-forming units/g, was similar for all methods evaluated. The lateral flow immunoassays evaluated could reliably detect S. enterica within 18 h, indicating that they may be useful for rapid point-of-care testing in equine practice applications. Additional evaluation is needed using samples from naturally infected cases and the environment to gain an accurate estimate of test sensitivity and specificity and to substantiate further the true value of these tests in clinical practice. © 2014 EVJ Ltd.

Preparation and properties of bio-compatible magnetic Fe3O4 nanoparticles

International Nuclear Information System (INIS)

Chan, H.T.; Do, Y.Y.; Huang, P.L.; Chien, P.L.; Chan, T.S.; Liu, R.S.; Huang, C.Y.; Yang, S.Y.; Horng, H.E.

2006-01-01

In this work, we study the preparation and properties of bio-compatible magnetic nanoparticles for immunoassay and DNA detection. The magnetite (Fe 3 O 4 ) nanoparticles were prepared by a chemical co-precipitation method and dextran was selected as the surfactant to suspend the nanoparticles. Suspended particles associated with avidin followed by biotin were qualitatively analyzed by enzyme-linked immunosorbent assay (ELISA) method. We found further the ethylenediamine blocked activated residual groups efficiently, hence enhancing the attachment of biotin for probing the avidin

Tumor specific lung cancer diagnostics with multiplexed FRET immunoassays

Science.gov (United States)

Geißler, D.; Hill, D.; Löhmannsröben, H.-G.; Thomas, E.; Lavigne, A.; Darbouret, B.; Bois, E.; Charbonnière, L. J.; Ziessel, R. F.; Hildebrandt, N.

2010-02-01

An optical multiplexed homogeneous (liquid phase) immunoassay based on FRET from a terbium complex to eight different fluorescent dyes is presented. We achieved highly sensitive parallel detection of four different lung cancer specific tumor markers (CEA, NSE, SCC and CYFRA21-1) within a single assay and show a proof-of-principle for 5- fold multiplexing. The method is well suited for fast and low-cost miniaturized point-of-care testing as well as for highthroughput screening in a broad range of in-vitro diagnostic applications.

A novel electrochemical sensor for lead ion based on cascade DNA and quantum dots amplification

International Nuclear Information System (INIS)

Tang, Shurong; Lu, Wei; Gu, Fang; Tong, Ping; Yan, Zhiming; Zhang, Lan

2014-01-01

A new enzyme-free and ultrasensitive electrochemical Pb 2+ biosensor was developed. By coupling the DNA-assisted cascade of hybridization reaction with the quantum dots (QDs) for signal amplification, a detection limit as low as 6.1 pM can be obtained for Pb 2+ . In this study, the “8-17” DNAzyme was used for specific recognition of Pb 2+ . In the presence of Pb 2+ , the DNAzyme was activated and cleaved the substrate strand. And then, the hybridization between the linker probe and signal probe was initiated, which resulted in formation of a long cascade DNA structure as well as assemble of numerous QDs at last. By the use of magnetic beads, the free signal probe can be easily removed by external magnetic field. After acid lysis, a great amount of redox cations can be released from the QDs and eventually result in significantly amplified electrochemical signals. This method is highly sensitive, selective and simple without the participation of any protein based enzyme (nuclease), thereby holds great potential for real sample analysis

Hydrogen storage material, electrochemically active material, electrochemical cell and electronic equipment

NARCIS (Netherlands)

2008-01-01

The invention relates to a hydrogen storage material comprising an alloy of magnesium. The invention further relates to an electrochemically active material and an electrochemical cell provided with at least one electrode comprising such a hydrogen storage material. Also, the invention relates to

Type I Clathrates as Novel Silicon Anodes: An Electrochemical and Structural Investigation

Science.gov (United States)

Li, Ying; Raghavan, Rahul; Wagner, Nicholas A.; Davidowski, Stephen K.; Baggetto, Loïc; Zhao, Ran; Cheng, Qian; Yarger, Jeffery L.; Veith, Gabriel M.; Ellis‐Terrell, Carol; Miller, Michael A.; Chan, Kwai S.

2015-01-01

Silicon clathrates contain cage‐like structures that can encapsulate various guest atoms or molecules. An electrochemical evaluation of type I silicon clathrates based on Ba8AlySi46−y as the anode material for lithium‐ion batteries is presented here. Postcycling characterization with nuclear magnetic resonance and X‐ray diffraction shows no discernible structural or volume changes even after electrochemical insertion of 44 Li (≈1 Li/Si) into the clathrate structure. The observed properties are in stark contrast with lithiation of other silicon anodes, which become amorphous and suffer from large volume changes. The electrochemical reactions are proposed to occur as single phase reactions at approximately 0.2 and 0.4 V versus Li/Li+ during lithiation and delithiation, respectively, distinct from diamond cubic or amorphous silicon anodes. Reversible capacities as high as 499 mAh g−1 at a 5 mA g−1 rate were observed for silicon clathrate with composition Ba8Al8.54Si37.46, corresponding to ≈1.18 Li/Si. These results show that silicon clathrates could be promising durable anodes for lithium‐ion batteries. PMID:27980951

Graphene quantum dots decorated with magnetic nanoparticles: Synthesis, electrodeposition, characterization and application as an electrochemical sensor towards determination of some amino acids at physiological pH

International Nuclear Information System (INIS)

Hasanzadeh, Mohammad; Karimzadeh, Ayub; Shadjou, Nasrin; Mokhtarzadeh, Ahad; Bageri, Leyla; Sadeghi, Sattar; Mahboob, Soltanali

2016-01-01

This study reports on the synthesis and characterization of a novel nano-composite, Fe 3 O 4 magnetic nanoparticles/graphene quantum dots (Fe 3 O 4 MNP-GQDs), for sensing of some amino acids. For the first time, as-synthesized GQDs and Fe 3 O 4 MNPs-GQDs was electrodeposited on the glassy carbon electrode (GCE) by cyclic voltammetry (CV) regime in the potential range from − 1.0 to 1.0 V. Fe 3 O 4 MNP-GQDs is engineered to specifically and effectively capture and enhancement the electrochemical signals of some amino acids at physiological pH due to the synergy among GQDs and magnetic nanoparticles. We have illustrated that the obtained Fe 3 O 4 MNPs-GQDs exhibited a much higher electroactivity individual GQDs and Fe 3 O 4 MNPs for the electrooxidation and detection of amino acid which was about 10 fold higher than for GQDs. Magnetic and specific properties of the Fe 3 O 4 MNP-GQDs can be exploited to capture and pre-concentration the amino acids onto its surface, which are important for detection of multi-amino acids. - Highlights: • Electrooxidation of amino acids was performed using Fe 3 O 4 MNP-GQDs. • Modified electrode shows new advantages as an amino acids sensor. • Excellent electrocatalytic activity was obtained for amino acids oxidation.

Graphene-Paper Based Electrochemical Sensors

DEFF Research Database (Denmark)

Zhang, Minwei; Halder, Arnab; Cao, Xianyi

2017-01-01

in electrochemical sensors and energy technologies amongothers. In this chapter, we present some examples to overview recent advances in theresearch and development of two-dimensional (2D) graphene papers as new materialsfor electrochemical sensors. The chapter covers the design, fabrication, functionalizationand...... functionalization ofgraphene papers with polymer and nanoscale functional building blocks for electrochemical-sensing purposes. In terms of electrochemical-sensing applications, the emphasis ison enzyme-graphene and nanoparticle-graphene paper-based systems for the detectionof glucose. We finally conclude...

Lateral Flow Immunoassays for Ebola Virus Disease Detection in Liberia.

Science.gov (United States)

Phan, Jill C; Pettitt, James; George, Josiah S; Fakoli, Lawrence S; Taweh, Fahn M; Bateman, Stacey L; Bennett, Richard S; Norris, Sarah L; Spinnler, David A; Pimentel, Guillermo; Sahr, Phillip K; Bolay, Fatorma K; Schoepp, Randal J

2016-10-15

Lateral flow immunoassays (LFIs) are point-of-care diagnostic assays that are designed for single use outside a formal laboratory, with in-home pregnancy tests the best-known example of these tests. Although the LFI has some limitations over more-complex immunoassay procedures, such as reduced sensitivity and the potential for false-positive results when using complex sample matrices, the assay has the benefits of a rapid time to result and ease of use. These benefits make it an attractive option for obtaining rapid results in an austere environment. In an outbreak of any magnitude, a field-based rapid diagnostic assay would allow proper patient transport and for safe burials to be conducted without the delay caused by transport of samples between remote villages and testing facilities. Use of such point-of-care instruments in the ongoing Ebola virus disease (EVD) outbreak in West Africa would have distinct advantages in control and prevention of local outbreaks, but proper understanding of the technology and interpretation of results are important. In this study, a LFI, originally developed by the Naval Medical Research Center for Ebola virus environmental testing, was evaluated for its ability to detect the virus in clinical samples in Liberia. Clinical blood and plasma samples and post mortem oral swabs submitted to the Liberian Institute for Biomedical Research, the National Public Health Reference Laboratory for EVD testing, were tested and compared to results of real-time reverse transcription-polymerase chain reaction (rRT-PCR), using assays targeting Ebola virus glycoprotein and nucleoprotein. The LFI findings correlated well with those of the real-time RT-PCR assays used as benchmarks. Rapid antigen-detection tests such as LFIs are attractive alternatives to traditional immunoassays but have reduced sensitivity and specificity, resulting in increases in false-positive and false-negative results. An understanding of the strengths, weaknesses, and

The antibody-based magnetic microparticle immunoassay using p-FET sensing platform for Alzheimer's disease pathogenic factor

Science.gov (United States)

Kim, Chang-Beom; Kim, Kwan-Soo; Song, Ki-Bong

2013-05-01

The importance of early Alzheimer's disease (AD) detection has been recognized to diagnose people at high risk of AD. The existence of intra/extracellular beta-amyloid (Aβ) of brain neurons has been regarded as the most archetypal hallmark of AD. The existing computed-image-based neuroimaging tools have limitations on accurate quantification of nanoscale Aβ peptides due to optical diffraction during imaging processes. Therefore, we propose a new method that is capable of evaluating a small amount of Aβ peptides by using photo-sensitive field-effect transistor (p-FET) integrated with magnetic force-based microbead collecting platform and selenium(Se) layer (thickness ~700 nm) as an optical filter. This method demonstrates a facile approach for the analysis of Aβ quantification using magnetic force and magnetic silica microparticles (diameter 0.2~0.3 μm). The microbead collecting platform mainly consists of the p-FET sensing array and the magnet (diameter ~1 mm) which are placed beneath each sensing region of the p-FET, which enables the assembly of the Aβ antibody conjugated microbeads, captures the Aβ peptides from samples, measures the photocurrents generated by the Q-dot tagged with Aβ peptides, and consequently results in the effective Aβ quantification.

Multi-scale magnetic nanoparticle based optomagnetic bioassay for sensitive DNA and bacteria detection

DEFF Research Database (Denmark)

Tian, Bo; Zardán Gómez De La Torre, Teresa; Donolato, Marco

2016-01-01

nanoparticles (binding to the target) and thus the optomagnetic response of the sample, which is measured by an optomagnetic setup including a 405 nm laser and a photodetector. The limit of detection is mainly set by the lowest measurable concentration of magnetic nanoparticles. Herein, as new results compared...... with the target. We show that the optimization and lowering of the 100 nm magnetic nanoparticle concentration result in a limit of detection of 780 fM of DNA coils formed by rolling circle amplification (size of about 1 μm) and 105 CFU per mL Salmonella (for immunoassay). These values are 15 times lower than...... those reported previously for this readout principle. Finally, we show that the 250 nm magnetic nanoparticles can serve as a second detection label for qualitative biplex detection of DNA coils formed by rolling circle amplification from V. cholerae and E. coli DNA coils using 100 nm and 250 nm magnetic...

Rational design of Raman-labeled nanoparticles for a dual-modality, light scattering immunoassay on a polystyrene substrate.

Science.gov (United States)

Israelsen, Nathan D; Wooley, Donald; Hanson, Cynthia; Vargis, Elizabeth

2016-01-01

Surface-enhanced Raman scattering (SERS) is a powerful light scattering technique that can be used for sensitive immunoassay development and cell labeling. A major obstacle to using SERS is the complexity of fabricating SERS probes since they require nanoscale characterization and optical uniformity. The light scattering response of SERS probes may also be modulated by the substrate used for SERS analysis. A typical SERS substrate such as quartz can be expensive. Polystyrene is a cheaper substrate option but can decrease the SERS response due to interfering Raman emission peaks and high background fluorescence. The goal of this research is to develop an optimized process for fabricating Raman-labeled nanoparticles for a SERS-based immunoassay on a polystyrene substrate. We have developed a method for fabricating SERS nanoparticle probes for use in a light scattering immunoassay on a polystyrene substrate. The light scattering profile of both spherical gold nanoparticle and gold nanorod SERS probes were characterized using Raman spectroscopy and optical absorbance spectroscopy. The effects of substrate interference and autofluorescence were reduced by selecting a Raman reporter with a strong light scattering response in a spectral region where interfering substrate emission peaks are minimized. Both spherical gold nanoparticles and gold nanorods SERS probes used in the immunoassay were detected at labeling concentrations in the low pM range. This analytical sensitivity falls within the typical dynamic range for direct labeling of cell-surface biomarkers using SERS probes. SERS nanoparticle probes were fabricated to produce a strong light scattering signal despite substrate interference. The optical extinction and inelastic light scattering of these probes was detected by optical absorbance spectroscopy and Raman spectroscopy, respectively. This immunoassay demonstrates the feasibility of analyzing strongly enhanced Raman signals on polystyrene, which is an

Silver nanoparticles deposited on graphene oxide for ultrasensitive surface-enhanced Raman scattering immunoassay of cancer biomarker.

Science.gov (United States)

Yang, Lin; Zhen, Shu Jun; Li, Yuan Fang; Huang, Cheng Zhi

2018-06-14

Graphene oxide (GO) exhibits distinctive Raman scattering features for its high frequency D (disordered) and tangential modes (G-band), which are characteristically sharp at 1580 cm-1 and 1350 cm-1, respectively, but are too weak for sensitive quantitation purposes. By depositing silver nanoparticles on the surface of GO in this contribution, both D and G bands of GO become enhanced. The enzyme label of this method controls the dissolution of silver nanoparticles on the surface of GO through hydrogen peroxide which is produced by the oxidation of the enzyme substrate. With the dissolution of the silver nanoparticles a greatly decreased SERS signal of GO was obtained. This strategy involves dual signal amplification of the enzyme and nanocomposites to improve the detection sensitivity. As a proof of concept, prostate specific antigen (PSA), a biomarker for prostate cancer, is successfully detected as a target by forming a sandwich structure in immunoassay. The SERS immunoassay possesses excellent analytical performance in the range 0.5 pg mL-1 to 500 pg mL-1 with a limit of detection of 0.23 pg mL-1, making the detection of PSA serum samples from prostate cancer patients satisfactory, demonstrating that the sensitive enzyme-assisted dissolved AgNPs SERS immunoassay of PSA has potential applications in clinical diagnosis.

Oriented antibody immobilization to polystyrene macrocarriers for immunoassay modified with hydrazide derivatives of poly(methacrylic acid

Directory of Open Access Journals (Sweden)

Vinokurova Ludmila G

2001-08-01

Full Text Available Abstract Background Hydrophobic polystyrene is the most common material for solid phase immunoassay. Proteins are immobilized on polystyrene by passive adsorption, which often causes considerable denaturation. Biological macromolecules were found to better retain their functional activity when immobilized on hydrophilic materials. Polyacrylamide is a common material for solid-phase carriers of biological macromolecules, including immunoreagents used in affinity chromatography. New macroformats for immunoassay modified with activated polyacrylamide derivatives seem to be promising. Results New polymeric matrices for immunoassay in the form of 0.63-cm balls which contain hydrazide functional groups on hydrophilic polymer spacer arms at their surface shell are synthesized by modification of aldehyde-containing polystyrene balls with hydrazide derivatives of poly(methacrylic acid. The beads contain up to 0.31 μmol/cm2 active hydrazide groups accessible for covalent reaction with periodate-oxidized antibodies. The matrices obtained allow carrying out the oriented antibody immobilization, which increases the functional activity of immunosorbents. Conclusions An efficient site-directed antibody immobilization on a macrosupport is realized. The polymer hydrophilic spacer arms are the most convenient and effective tools for oriented antibody coupling with molded materials. The suggested scheme can be used for the modification of any other solid supports containing electrophilic groups reacting with hydrazides.

Aptamer-conjugated silver nanoparticles for electrochemical dual-aptamer-based sandwich detection of staphylococcus aureus.

Science.gov (United States)

Abbaspour, Abdolkarim; Norouz-Sarvestani, Fatemeh; Noori, Abolhassan; Soltani, Noushin

2015-06-15

Staphylococcus aureus (S. aureus) is one of the most important human pathogens and causes numerous illnesses. In this study, we report a sensitive and highly selective dual-aptamer-based sandwich immunosensor for the detection of S. aureus. In this bioassay system, a biotinylated primary anti-S.aureus aptamer was immobilized on streptavidin coated magnetic beads (MB), which serves as a capture probe. A secondary anti-S.aureus aptamer was conjugated to silver nanoparticles (Apt-AgNP) that sensitively reports the detection of the target. In the presence of target bacterium, an Apt/S.aureus/apt-AgNP sandwich complex is formed on the MB surface and the electrochemical signal of AgNPs followed through anodic stripping voltammetry. The proposed sandwich assay benefits from advantageous of a sandwich assay for increased specificity, MB as carriers of affinity ligands for solution-phase recognition and fast magnetic separation, AgNPs for signal amplification, and an electrochemical stripping voltammetry read-out as a simple and sensitive detection. The electrochemical immunosensor shows an extended dynamic range from 10 to 1×10(6) cfu/mL with a low detection limit of 1.0 cfu/mL (S/N=3). Furthermore, the possible interference of other analog bacteria was studied. To assess the general applicability of this sensor, we investigated the quantification of S. aureus in real water samples. The results were compared to the experimental results obtained from a plate counting method, which demonstrated an acceptable consistency. Copyright © 2014 Elsevier B.V. All rights reserved.

Electrochemical force microscopy

Energy Technology Data Exchange (ETDEWEB)

Kalinin, Sergei V.; Jesse, Stephen; Collins, Liam F.; Rodriguez, Brian J.

2017-01-10

A system and method for electrochemical force microscopy are provided. The system and method are based on a multidimensional detection scheme that is sensitive to forces experienced by a biased electrode in a solution. The multidimensional approach allows separation of fast processes, such as double layer charging, and charge relaxation, and slow processes, such as diffusion and faradaic reactions, as well as capturing the bias dependence of the response. The time-resolved and bias measurements can also allow probing both linear (small bias range) and non-linear (large bias range) electrochemical regimes and potentially the de-convolution of charge dynamics and diffusion processes from steric effects and electrochemical reactivity.

A competitive enzyme immunoassay for the quantitative detection of cocaine from banknotes and latent fingermarks.

Science.gov (United States)

van der Heide, Susan; Garcia Calavia, Paula; Hardwick, Sheila; Hudson, Simon; Wolff, Kim; Russell, David A

2015-05-01

A sensitive and versatile competitive enzyme immunoassay (cEIA) has been developed for the quantitative detection of cocaine in complex forensic samples. Polyclonal anti-cocaine antibody was purified from serum and deposited onto microtiter plates. The concentration of the cocaine antibody adsorbed onto the plates, and the dilution of the cocaine-HRP hapten were both studied to achieve an optimised immunoassay. The method was successfully used to quantify cocaine in extracts taken from both paper currency and latent fingermarks. The limit of detection (LOD) of 0.162ngmL(-1) achieved with the assay compares favourably to that of conventional chromatography-mass spectroscopy techniques, with an appropriate sensitivity for the quantification of cocaine at the low concentrations present in some forensic samples. The cEIA was directly compared to LC-MS for the analysis of ten UK banknote samples. The results obtained from both techniques were statistically similar, suggesting that the immunoassay was unaffected by cross-reactivity with potentially interfering compounds. The cEIA was used also for the detection of cocaine in extracts from latent fingermarks. The results obtained were compared to the cocaine concentrations detected in oral fluid sampled from the same individual. Using the cEIA, we have shown, for the first time, that endogeneously excreted cocaine can be detected and quantified from a single latent fingermark. Additionally, it has been shown that the presence of cocaine, at similar concentrations, in more than one latent fingermark from the same individual can be linked with those concentrations found in oral fluid. These results show that detection of drugs in latent fingermarks could directly indicate whether an individual has consumed the drug. The specificity and feasibility of measuring low concentrations of cocaine in complex forensic samples demonstrate the effectiveness and robustness of the assay. The immunoassay presents a simple and cost

False-negative syphilis treponemal enzyme immunoassay results in an HIV-infected case-patient.

Science.gov (United States)

Katz, Alan R; Komeya, Alan Y; Tomas, Juval E

2017-06-01

We present a case report of a false-negative syphilis treponemal enzyme immunoassay test result in an HIV-infected male. While treponemal tests are widely considered to be more sensitive and specific than non-treponemal tests, our findings point to potential challenges using the reverse sequence syphilis screening algorithm.

Upconverting nanophosphors as reporters in a highly sensitive heterogeneous immunoassay for cardiac troponin I

Energy Technology Data Exchange (ETDEWEB)

Sirkka, Nina, E-mail: nkelon@utu.fi; Lyytikäinen, Annika; Savukoski, Tanja; Soukka, Tero

2016-06-21

Photon upconverting nanophosphors (UCNPs) have a unique capability to produce anti-Stokes emission at visible wavelengths via sequential multiphoton absorption upon infrared excitation. Since the anti-Stokes emission can be easily spectrally resolved from the Stokes' shifted autofluorescence, the upconversion luminescence (UCL) is a highly attractive reporter technology for optical biosensors and biomolecular binding assays – potentially enabling unprecedented sensitivity in separation-based solid-phase immunoassays. UCL technology has not previously been applied in sensitive detection of cardiac troponin I (cTnI), which requires highly sensitive detection to enable accurate and timely diagnosis of myocardial infarction. We have developed an UCL-based immunoassay for cTnI using NaYF{sub 4}: Yb{sup 3+}, Er{sup 3+} UCNPs as reporters. Biotinylated anti-cTnI monoclonal antibody (Mab) and Fab fragment immobilized to streptavidin-coated wells were used to capture cTnI. Captured cTnI was detected from dry well surface after a 15 min incubation with poly(acrylic acid) coated UCNPs conjugated to second anti-cTnI Mab. UCL was measured with a dedicated UCL microplate reader. The UCL-based immunoassay allowed sensitive detection of cTnI. The limit of detection was 3.14 ng L{sup −1}. The calibration curve was linear up to cTnI concentration 50,000 ng L{sup −1}. Plasma recoveries of added cTnI were 92–117%. Obtained cTnI concentrations from five normal plasma samples were 4.13–10.7 ng L{sup −1} (median 5.06 ng L{sup −1}). There is yet significant potential for even further improved limit of detection by reducing non-specifically bound fraction of the Mab-conjugated UCNPs. The assay background with zero calibrator was over 40-fold compared to the background obtained from wells where the reporter conjugate had been excluded. - Highlights: • Detection of attomole analyte quantity in microwell using upconversion luminescence. • Upconverting

Dot immunoassay for the simultaneous determination of postvaccination immunity against pertussis, diphtheria, and tetanus.

Science.gov (United States)

Khramtsov, Pavel; Bochkova, Maria; Timganova, Valeria; Zamorina, Svetlana; Rayev, Mikhail

2017-06-01

A dot immunoassay for simultaneous semiquantitative detection of IgG against tetanus toxoid (Ttx) and diphtheria toxoid (Dtx) and qualitative detection of anti-Bordetella pertussis IgGs in human blood serum using carbon nanoparticles functionalized with streptococcal protein G was developed. Inactivated B. pertussis cells in suspension form were used as an antigen in the immunoassay. Pertussis, tetanus, and diphtheria antigens were separately spotted onto nitrocellulose strips, and then the immunostrips were successively incubated with blood sera and a suspension of carbon nanoparticles. The immunostrips were then scanned with a flatbed scanner, and the images obtained were processed with ImageJ. One hundred fifty-five venous blood serum samples from children vaccinated with diphtheria, tetanus, and whole-cell pertussis (DTwP) vaccine were tested in comparison with a conventional ELISA and agglutination test. The total time required for analysis of 32 serum samples was less than 3 h. Comparison between the results of the dot immunoassay and the corresponding ELISA/agglutination test revealed a high level of agreement (Cohen's kappa between 0.765 and 0.813). The lower limit of quantification was 0.06 IU/ml for anti-Ttx and anti-Dtx. The intra-assay coefficients of variation were less than 15% for anti-Ttx and anti-Dtx and less than 10% for anti-pertussis. The diagnostic sensitivity of detection of the antibody protection level was 93.5% for anti-Ttx [95% confidence interval (CI) 83.5-97.9%], 92.4% for anti-Dtx (95% CI 80.9297.5%), and 90.2% for anti-pertussis (95% CI 75.9-96.8%). The diagnostic specificity was 90.9% for anti-Ttx (95% CI 57.1-99.5%), 85% for anti-Dtx (95% CI 61.1-96.0%), and 89.3% for anti-pertussis (95%CI 80.8-94.5%). The dot immunoassay developed does not require expensive reading equipment, and allows detection of antibodies against three antigens in a single analysis. The immunostrips can be stored for a long time without changes in the

Determination of phospholipid transfer proteins in rat tissues by immunoassays

International Nuclear Information System (INIS)

Teerlink, T.

1983-01-01

Several quantitative immunoassays have been developed for two phospholipid transfer proteins from rat liver, i.e. the phosphatidylcholine transfer protein and the non-specific lipid transfer protein. The development of a double-antibody radioimmunoassay for the phosphatidylcholine transfer protein is described. The transfer protein was labelled with iodine-125 by the mild glucose oxidase-lactoperoxidase method. Although less than one tyrosine residue per molecule of transfer protein was labelled, only 20% of the labelled transfer protein was immunoprecipitable. This value could be increased to 80% by purifying the labelled protein by affinity chromatography on a column of anti-phosphatidylcholine transfer protein-IgG coupled to Sepharose 4B. The radioimmunoassay was used to determine the levels of phosphatidylcholine transfer protein in homogenates and 105 000 xg supernatants from various rat tissues as well as several Morris hepatomas. An enzyme immunoassay for the non-specific lipid transfer protein is also described. The antiserum that was raised especially by the author was cross-reactive with the non-specific lipid transfer protein present in 105 000 xg supernatants from human, mouse and bovine liver. The non-specific lipid transfer protein lost its immunoreactivity upon labelling with iodine-125 using different labelling techniques. Therefore, a regular radioimmunoassay could not be developed. The results of these different assays were compared. (Auth.)

Materials for electrochemical capacitors

Science.gov (United States)

Simon, Patrice; Gogotsi, Yury

2008-11-01

Electrochemical capacitors, also called supercapacitors, store energy using either ion adsorption (electrochemical double layer capacitors) or fast surface redox reactions (pseudo-capacitors). They can complement or replace batteries in electrical energy storage and harvesting applications, when high power delivery or uptake is needed. A notable improvement in performance has been achieved through recent advances in understanding charge storage mechanisms and the development of advanced nanostructured materials. The discovery that ion desolvation occurs in pores smaller than the solvated ions has led to higher capacitance for electrochemical double layer capacitors using carbon electrodes with subnanometre pores, and opened the door to designing high-energy density devices using a variety of electrolytes. Combination of pseudo-capacitive nanomaterials, including oxides, nitrides and polymers, with the latest generation of nanostructured lithium electrodes has brought the energy density of electrochemical capacitors closer to that of batteries. The use of carbon nanotubes has further advanced micro-electrochemical capacitors, enabling flexible and adaptable devices to be made. Mathematical modelling and simulation will be the key to success in designing tomorrow's high-energy and high-power devices.

Electrochemical reduction of NOx

DEFF Research Database (Denmark)

Traulsen, Marie Lund

NO and NO2 (collectively referred to as NOx) are air pollutants, and the largest single contributor to NOx pollution is automotive exhaust. This study investigates electrochemical deNOx, a technology which aims to remove NOx from automotive diesel exhaust by electrochemical reduction of NOx to N2...... and O2. The focus in this study is on improving the activity and selectivity of solid oxide electrodes for electrochemical deNOx by addition of NOx storage compounds to the electrodes. Two different composite electrodes, La0.85Sr0.15MnO3-δ-Ce0.9Gd0.1O1.95 (LSM15-CGO10) and La0.85Sr0.15FeO3-δ-Ce0.9Gd0.1O......1.95 (LSF15-CGO10), have been investigated in combination with three different NOx storage compounds: BaO, K2O and MnOx. The main focus in the investigation has been on conversion measurements and electrochemical characterization, the latter by means of electrochemical impedance spectroscopy...

In-electrode vs. on-electrode: ultrasensitive Faraday cage-type electrochemiluminescence immunoassay.

Science.gov (United States)

Guo, Zhiyong; Sha, Yuhong; Hu, Yufang; Wang, Sui

2016-03-28

A new-concept of an "in-electrode" Faraday cage-type electrochemiluminescence immunoassay (ECLIA) method for the ultrasensitive detection of neurotensin (NT) was reported with capture antibody (Ab1)-nanoFe3O4@graphene (GO) and detector antibody (Ab2)&N-(4-aminobutyl)-N-ethylisoluminol (ABEI)@GO, which led to about 1000-fold improvement in sensitivity by extending the Helmholtz plane (OHP) of the proposed electrode assembly effectively.

Electrochemical oxidation of cholesterol in acetonitrile leads to the formation of cholesta-4,6-dien-3-one

International Nuclear Information System (INIS)

Hosokawa, Yu-Ya; Hakamata, Hideki; Murakami, Tomonori; Aoyagi, Sakae; Kuroda, Minpei; Mimaki, Yoshihiro; Ito, Ayumi; Morosawa, Sayaka; Kusu, Fumiyo

2009-01-01

Cholesterol was shown to be oxidized at the glassy carbon electrode in an acetonitrile-2-propanol mixture and this oxidation reaction was applied to the determination of serum total cholesterol by high-performance liquid chromatography with electrochemical detection (K. Hojo, H. Hakamata, A. Ito, A. Kotani, C. Furukawa, Y.Y. Hosokawa, F. Kusu, J. Chromatogr. A 1166 (2007) 135-141). To gain insight into the detection mechanisms of cholesterol, an electrolytic product of cholesterol was collected and characterized by infrared spectroscopy, one- and two-dimensional nuclear magnetic resonance, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The three techniques, together with comparisons of literature spectral data, confirmed the formation of cholesta-4,6-dien-3-one. The conversion of cholesterol to cholesta-4,6-dien-3-one, a four-electron, four-proton electrochemical process, has been proposed as an electrochemical oxidation mechanism of cholesterol in acetonitrile.

An anti-phospholipase A2 receptor quantitative immunoassay and epitope analysis in membranous nephropathy reveals different antigenic domains of the receptor.

Directory of Open Access Journals (Sweden)

Astrid Behnert

Full Text Available The phospholipase A2 receptor (PLA2R was recently discovered as a target autoantigen in patients with idiopathic membranous nephropathy (IMN. Published evidence suggests that the autoantibodies directed towards a conformation dependent epitope are currently effectively detected by a cell based assay (CBA utilizing indirect immunofluorescence (IIF on tissue culture cells transfected with the PLA2R cDNA. Limitations of such IIF-CBA assays include observer dependent subjective evaluation of semi-quantitative test results and the protocols are not amenable to high throughput diagnostic testing. We developed a quantitative, observer independent, high throughput capture immunoassay for detecting PLA2R autoantibodies on an addressable laser bead immunoassay (ALBIA platform. Since reactive domains of PLA2R (i.e. epitopes could be used to improve diagnostic tests by using small peptides in various high throughput diagnostic platforms, we identified PLA2R epitopes that bound autoantibodies of IMN patients. These studies confirmed that inter-molecular epitope spreading occurs in IMN but use of the cognate synthetic peptides in immunoassays was unable to conclusively distinguish between IMN patients and normal controls. However, combinations of these peptides were able to effectively absorb anti-PLA2R reactivity in IIF-CBA and an immunoassay that employed a lysate derived from HEK cells tranfected with and overexpressing PLA2R. While we provide evidence of intermolecular epitope spreading, our data indicates that in addition to conformational epitopes, human anti-PLA2R reactivity in a commercially available CBA and an addressable laser bead immunoassay is significantly absorbed by peptides representing epitopes of PLA2R.

Electrochemically deposited hybrid nickel-cobalt hexacyanoferrate nanostructures for electrochemical supercapacitors

International Nuclear Information System (INIS)

Safavi, A.; Kazemi, S.H.; Kazemi, H.

2011-01-01

Highlights: → Nanostructured hybrid nickel-cobalt hexacyanoferrate is used in supercapacitors. → A high capacitance (765 F g -1 ) is obtained at a specific current of 0.2 A g -1 . → Long cycle-life and excellent stability are demonstrated during 1000 cycles. - Abstract: This study describes the use of electrodeposited nanostructured hybrid nickel-cobalt hexacyanoferrate in electrochemical supercapacitors. Herein, various compositions of nickel and cobalt hexacyanoferrates (Ni/CoHCNFe) nanostructures are electrodeposited on an inexpensive stainless steel substrate using cyclic voltammetric (CV) method. The morphology of the electrodeposited nanostructures is studied using scanning electron microscopy, while their electrochemical characterizations are investigated using CV, galvanostatic charge and discharge and electrochemical impedance spectroscopy. The results show that the nanostructures of hybrid metal cyanoferrate, shows a much higher capacitance (765 F g -1 ) than those obtained with just nickel hexacyanoferrate (379 F g -1 ) or cobalt hexacyanoferrate (277 F g -1 ). Electrochemical impedance spectroscopy results confirm the favorable capacitive behavior of the electrodeposited materials. The columbic efficiency is approximately 95% based on the charge and discharge experiments. Long cycle-life and excellent stability of the nanostructured materials are also demonstrated during 1000 cycles.

Electrochemically deposited hybrid nickel-cobalt hexacyanoferrate nanostructures for electrochemical supercapacitors

Energy Technology Data Exchange (ETDEWEB)

Safavi, A., E-mail: safavi@chem.susc.ac.ir [Department of Chemistry, College of Sciences, Shiraz University, Shiraz 71454 (Iran, Islamic Republic of); Nanotechnology Research Institute, Shiraz University, Shiraz (Iran, Islamic Republic of); Kazemi, S.H., E-mail: habibkazemi@iasbs.ac.ir [Department of Chemistry, Institute for Advanced Studies in Basic Sciences (IASBS), Zanjan 45137-66731 (Iran, Islamic Republic of); Kazemi, H. [Department of Chemistry, College of Sciences, Shiraz University, Shiraz 71454 (Iran, Islamic Republic of)

2011-10-30

Highlights: > Nanostructured hybrid nickel-cobalt hexacyanoferrate is used in supercapacitors. > A high capacitance (765 F g{sup -1}) is obtained at a specific current of 0.2 A g{sup -1}. > Long cycle-life and excellent stability are demonstrated during 1000 cycles. - Abstract: This study describes the use of electrodeposited nanostructured hybrid nickel-cobalt hexacyanoferrate in electrochemical supercapacitors. Herein, various compositions of nickel and cobalt hexacyanoferrates (Ni/CoHCNFe) nanostructures are electrodeposited on an inexpensive stainless steel substrate using cyclic voltammetric (CV) method. The morphology of the electrodeposited nanostructures is studied using scanning electron microscopy, while their electrochemical characterizations are investigated using CV, galvanostatic charge and discharge and electrochemical impedance spectroscopy. The results show that the nanostructures of hybrid metal cyanoferrate, shows a much higher capacitance (765 F g{sup -1}) than those obtained with just nickel hexacyanoferrate (379 F g{sup -1}) or cobalt hexacyanoferrate (277 F g{sup -1}). Electrochemical impedance spectroscopy results confirm the favorable capacitive behavior of the electrodeposited materials. The columbic efficiency is approximately 95% based on the charge and discharge experiments. Long cycle-life and excellent stability of the nanostructured materials are also demonstrated during 1000 cycles.

Detection of carcinoembryonic antigen using functional magnetic and fluorescent nanoparticles in magnetic separators

Energy Technology Data Exchange (ETDEWEB)

Tsai, H. Y., E-mail: annetsai@csmu.edu.tw [Chung Shan Medical University, Department of Applied Chemistry (China); Chang, C. Y.; Li, Y. C.; Chu, W. C.; Viswanathan, K.; Bor Fuh, C., E-mail: cbfuh@ncnu.edu.tw [National Chi Nan University, Department of Applied Chemistry (China)

2011-06-15

We combined a sandwich immunoassay, anti-CEA/CEA/anti-CEA, with functional magnetic ({approx}80 nm) and fluorescent ({approx}180 nm) nanoparticles in magnetic separators to demonstrate a detection method for carcinoembryonic antigen (CEA). Determination of CEA in serum can be used in clinical diagnosis and monitoring of tumor-related diseases. The CEA concentrations in samples were deduced and determined based on the reference plot using the measured fluorescent intensity of sandwich nanoparticles from the sample. The linear range of CEA detection was from 18 ng/mL to 1.8 pg/mL. The detection limit of CEA was 1.8 pg/mL. In comparison with most other detection methods, this method had advantages of lower detection limit and wider linear range. The recovery was higher than 94%. The CEA concentrations of two serum samples were determined to be 9.0 and 55 ng/mL, which differed by 6.7% (9.6 ng/mL) and 9.1% (50 ng/mL) from the measurements of enzyme-linked immunosorbent assay (ELISA), respectively. The analysis time can be reduced to one third of ELISA. This method has good potential for other biomarker detections and biochemical applications.

Analytical evaluation of the novel Lumipulse G BRAHMS procalcitonin immunoassay

OpenAIRE

Ruzzenente, Orazio; Salvagno, Gian Luca; Gelati, Matteo; Lippi, Giuseppe

2016-01-01

Objectives: This study was designed to evaluate the analytical performance of the novel Lumipulse G1200 BRAHMS procalcitonin (PCT) immunoassay. Design and methods: This analytical evaluation encompassed the calculation of the limit of blank (LOB), limit of detection (LOD), functional sensitivity, intra- and inter-assay imprecision, confirmation of linearity and a comparison with the Vidas BRAHMS PCT assay. Results: The LOB, LOD and functional sensitivity were 0.0010 ng/mL, 0.0016 ng/mL and ...

Radio-immunoassays for glucagon-like peptides 1 and 2 (GLP-1 and GLP-2)

DEFF Research Database (Denmark)

Orskov, C; Holst, J J

1987-01-01

Gene-sequencing studies have shown that the glucagon precursor contains two additional glucagon-like sequences, the so-called glucagon-like peptides 1 and 2 (GLP-1 and GLP-2). We developed radio-immunoassays against synthetic peptides corresponding to these sequences. Antisera were raised in rabb...

Monoclonal antibody-based broad-specificity immunoassay for monitoring organophosphorus pesticides in environmental water samples

Science.gov (United States)

The extensive use of organophosphorus pesticides (OPs) in agriculture and domestic settings can result in widespread water contamination. The development of easy-to-use and rapid-screening immunoassay methods in a class-selective manner is a topic of considerable environmental interest. In this wo...

Electrochemical Sensors for Clinic Analysis

Directory of Open Access Journals (Sweden)

Guang Li

2008-03-01

Full Text Available Demanded by modern medical diagnosis, advances in microfabrication technology have led to the development of fast, sensitive and selective electrochemical sensors for clinic analysis. This review addresses the principles behind electrochemical sensor design and fabrication, and introduces recent progress in the application of electrochemical sensors to analysis of clinical chemicals such as blood gases, electrolytes, metabolites, DNA and antibodies, including basic and applied research. Miniaturized commercial electrochemical biosensors will form the basis of inexpensive and easy to use devices for acquiring chemical information to bring sophisticated analytical capabilities to the non-specialist and general public alike in the future.

Electrochemical biosensors for hormone analyses.

Science.gov (United States)

Bahadır, Elif Burcu; Sezgintürk, Mustafa Kemal

2015-06-15

Electrochemical biosensors have a unique place in determination of hormones due to simplicity, sensitivity, portability and ease of operation. Unlike chromatographic techniques, electrochemical techniques used do not require pre-treatment. Electrochemical biosensors are based on amperometric, potentiometric, impedimetric, and conductometric principle. Amperometric technique is a commonly used one. Although electrochemical biosensors offer a great selectivity and sensitivity for early clinical analysis, the poor reproducible results, difficult regeneration steps remain primary challenges to the commercialization of these biosensors. This review summarizes electrochemical (amperometric, potentiometric, impedimetric and conductometric) biosensors for hormone detection for the first time in the literature. After a brief description of the hormones, the immobilization steps and analytical performance of these biosensors are summarized. Linear ranges, LODs, reproducibilities, regenerations of developed biosensors are compared. Future outlooks in this area are also discussed. Copyright © 2014 Elsevier B.V. All rights reserved.

Factitious Graves' Disease Due to Biotin Immunoassay Interference-A Case and Review of the Literature.

Science.gov (United States)

Elston, Marianne S; Sehgal, Shekhar; Du Toit, Stephen; Yarndley, Tania; Conaglen, John V

2016-09-01

Biotin (vitamin B7) is an essential co-factor for four carboxylases involved in fatty acid metabolism, leucine degradation, and gluconeogenesis. The recommended daily intake (RDI) of biotin is approximately 30 μg per day. Low-moderate dose biotin is a common component of multivitamin preparations, and high-dose biotin (10 000 times RDI) has been reported to improve clinical outcomes and quality of life in patients with progressive multiple sclerosis. Biotin is also a component of immunoassays, and supplementation may cause interference in both thyroid and non-thyroid immunoassays. To assess whether biotin ingestion caused abnormal thyroid function tests (TFTs) in a patient through assay interference. We report a patient with biotin-associated abnormal TFTs and a systematic review of the literature. A tertiary endocrine service in Hamilton, New Zealand. The patient had markedly abnormal TFTs that did not match the clinical context. After biotin cessation, TFTs normalized far more rapidly than possible given the half-life of T4, consistent with assay interference by biotin. Multiple other analytes also tested abnormal in the presence of biotin. Biotin ingested in moderate to high doses can cause immunoassay interference. Depending on the assay format, biotin interference can result in either falsely high or low values. Interference is not limited to thyroid tests and has the potential to affect a wide range of analytes. It is important for clinicians to be aware of this interaction to prevent misdiagnosis and inappropriate treatment.

Magnetic Bead Based Immunoassay for Autonomous Detection of Toxins

Energy Technology Data Exchange (ETDEWEB)

Kwon, Y; Hara, C A; Knize, M G; Hwang, M H; Venkatesteswaran, K S; Wheeler, E K; Bell, P M; Renzi, R F; Fruetel, J A; Bailey, C G

2008-05-01

As a step towards toward the development of a rapid, reliable analyzer for bioagents in the environment, we are developing an automated system for the simultaneous detection of a group of select agents and toxins. To detect toxins, we modified and automated an antibody-based approach previously developed for manual medical diagnostics that uses fluorescent eTag{trademark} reporter molecules and is suitable for highly multiplexed assays. Detection is based on two antibodies binding simultaneously to a single antigen, one of which is labeled with biotin while the other is conjugated to a fluorescent eTag{trademark} through a cleavable linkage. Aqueous samples are incubated with the mixture of antibodies along with streptavidin-coated magnetic beads coupled to a photo-activatable porphyrin complex. In the presence of antigen, a molecular complex is formed where the cleavable linkage is held in proximity to the photoactivable group. Upon excitation at 680 nm, free radicals are generated, which diffuse and cleave the linkage, releasing the eTags{trademark}. Released eTags{trademark} are analyzed using capillary gel electrophoresis with laser-induced fluorescence detection. Limits of detection for ovalbumin and botulinum toxoid individually were 4 ng/mL (or 80 pg) and 16 ng/mL (or 320 pg), respectively, using the manual assay. In addition, we demonstrated the use of pairs of antibodies from different sources in a single assay to decrease the rate of false positives. Automation of the assay was demonstrated on a flow-through format with higher LODs of 125 ng/mL (or 2.5 ng) each of a mixture of ovalbumin and botulinum toxoid. This versatile assay can be easily modified with the appropriate antibodies to detect a wide range of toxins and other proteins.

Graphene quantum dots decorated with magnetic nanoparticles: Synthesis, electrodeposition, characterization and application as an electrochemical sensor towards determination of some amino acids at physiological pH

Energy Technology Data Exchange (ETDEWEB)

Hasanzadeh, Mohammad, E-mail: mhmmd_hasanzadeh@yahoo.com [Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz 51664 (Iran, Islamic Republic of); Pharmaceutical Analysis Research Center, Tabriz University of Medical Sciences, Tabriz 51664 (Iran, Islamic Republic of); Karimzadeh, Ayub [Department of Biochemistry, Higher Education Institute of Rab-Rashid, Tabriz (Iran, Islamic Republic of); School of Medicine, Gonabad University of Medical Sciences, Gonabad (Iran, Islamic Republic of); Shadjou, Nasrin [Department of Nanochemistry, Nano Technology Research Center, Urmia University, Urmia 57154 (Iran, Islamic Republic of); Department of Nano Technology, Faculty of Science, Urmia University, Urmia 57154 (Iran, Islamic Republic of); Mokhtarzadeh, Ahad [School of Medicine, Gonabad University of Medical Sciences, Gonabad (Iran, Islamic Republic of); Department of Biochemistry, Higher Education Institute of Rab-Rashid, Tabriz (Iran, Islamic Republic of); Bageri, Leyla [Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz 51664 (Iran, Islamic Republic of); Sadeghi, Sattar; Mahboob, Soltanali [Department of Biochemistry, Higher Education Institute of Rab-Rashid, Tabriz (Iran, Islamic Republic of)

2016-11-01

This study reports on the synthesis and characterization of a novel nano-composite, Fe{sub 3}O{sub 4} magnetic nanoparticles/graphene quantum dots (Fe{sub 3}O{sub 4} MNP-GQDs), for sensing of some amino acids. For the first time, as-synthesized GQDs and Fe{sub 3}O{sub 4} MNPs-GQDs was electrodeposited on the glassy carbon electrode (GCE) by cyclic voltammetry (CV) regime in the potential range from − 1.0 to 1.0 V. Fe{sub 3}O{sub 4} MNP-GQDs is engineered to specifically and effectively capture and enhancement the electrochemical signals of some amino acids at physiological pH due to the synergy among GQDs and magnetic nanoparticles. We have illustrated that the obtained Fe{sub 3}O{sub 4} MNPs-GQDs exhibited a much higher electroactivity individual GQDs and Fe{sub 3}O{sub 4} MNPs for the electrooxidation and detection of amino acid which was about 10 fold higher than for GQDs. Magnetic and specific properties of the Fe{sub 3}O{sub 4} MNP-GQDs can be exploited to capture and pre-concentration the amino acids onto its surface, which are important for detection of multi-amino acids. - Highlights: • Electrooxidation of amino acids was performed using Fe{sub 3}O{sub 4} MNP-GQDs. • Modified electrode shows new advantages as an amino acids sensor. • Excellent electrocatalytic activity was obtained for amino acids oxidation.

Effect of Amine Adlayer on Electrochemical Uric Acid Sensor Conducted on Electrochemically Reduced Graphene Oxide

Energy Technology Data Exchange (ETDEWEB)

Park, Sumi; Kim, Kyuwon [Incheon National University, Incheon (Korea, Republic of)

2016-03-15

The electrochemical biosensing efficiency of uric acid (UA) detection on an electrochemically reduced graphene oxide (ERGO)-decorated electrode surface was studied by using various amine linkers used to immobilize ERGO. The amine linkers aminoethylphenyldiazonium , 2,2'-(ethylenedioxy)bis(ethylamine), 3-aminopro-pyltriethoxysilane, and polyethyleneimine were coated on indium-tin-oxide electrode surfaces through chemical or electrochemical deposition methods. ERGO-decorated surfaces were prepared by the electrochemical reduction of graphene oxide (GO), which was immobilized on the amine-coated electrode surfaces through the electrostatic interaction between GO and the ammonium ion of the linker on the surface. We monitored the sensing results of electrochemical UA detection with differential pulse voltammetry. The ERGO-modified surface presented electrocatalytic oxidation of UA and ascorbic acid. Among the different amines tested, 3-aminopropyltriethoxysilane provided the best biosensing performance in terms of sensitivity and reproducibility.

Effect of Amine Adlayer on Electrochemical Uric Acid Sensor Conducted on Electrochemically Reduced Graphene Oxide

International Nuclear Information System (INIS)

Park, Sumi; Kim, Kyuwon

2016-01-01

The electrochemical biosensing efficiency of uric acid (UA) detection on an electrochemically reduced graphene oxide (ERGO)-decorated electrode surface was studied by using various amine linkers used to immobilize ERGO. The amine linkers aminoethylphenyldiazonium , 2,2'-(ethylenedioxy)bis(ethylamine), 3-aminopro-pyltriethoxysilane, and polyethyleneimine were coated on indium-tin-oxide electrode surfaces through chemical or electrochemical deposition methods. ERGO-decorated surfaces were prepared by the electrochemical reduction of graphene oxide (GO), which was immobilized on the amine-coated electrode surfaces through the electrostatic interaction between GO and the ammonium ion of the linker on the surface. We monitored the sensing results of electrochemical UA detection with differential pulse voltammetry. The ERGO-modified surface presented electrocatalytic oxidation of UA and ascorbic acid. Among the different amines tested, 3-aminopropyltriethoxysilane provided the best biosensing performance in terms of sensitivity and reproducibility.

Development of a simple extraction procedure for chlorpyrifos determination in food samples by immunoassay.

Science.gov (United States)

Gabaldón, J A; Maquieira, A; Puchades, R

2007-02-28

The suitability of immunoassay methodology for rapid and accurate determination of chlorpyrifos in vegetables was tested. The optimised ELISA detection limit was 0.32ng/ml, with a working range from 0.69 to 6.21ng/ml and an immunoassay test-mid point (IC(50)) of 2.08ng/ml. A rapid sample preparation procedure considering different parameters such as the amount of sample, volume of extractant, extraction time and dilution factor was optimised. The developed direct extraction (DE) and multiresidue (ME) standard procedures were performed in different fortified fresh and processed vegetable samples (tomato, bonnet pepper, bean, pea, asparagus, broccoli, watermelon, melon, lettuce, cucumber, celery and red pepper). Recoveries were in all cases in the whole range 85.2-108.9% for both DE and ME extracts. Also, the comparison of the results obtained by both immunochemical and chromatographic methods for spiked fruits and vegetables were good with a correlation coefficient (r) of 0.97.

Accurate Point-of-Care Detection of Ruptured Fetal Membranes: Improved Diagnostic Performance Characteristics with a Monoclonal/Polyclonal Immunoassay

Directory of Open Access Journals (Sweden)

Linda C. Rogers

2016-01-01

Full Text Available Objective Accurate and timely diagnosis of rupture of membranes (ROM is imperative to allow for gestational age-specific interventions. This study compared the diagnostic performance characteristics between two methods used for the detection of ROM as measured in the same patient. Methods Vaginal secretions were evaluated using the conventional fern test as well as a point-of-care monoclonal/polyclonal immunoassay test (ROM Plus® in 75 pregnant patients who presented to labor and delivery with complaints of leaking amniotic fluid. Both tests were compared to analytical confirmation of ROM using three external laboratory tests. Diagnostic performance characteristics were calculated including sensitivity, specificity, positive predictive value (PPV, negative predictive value (NPV, and accuracy. Results Diagnostic performance characteristics uniformly favored ROM detection using the immunoassay test compared to the fern test: sensitivity (100% vs. 77.8%, specificity (94.8% vs. 79.3%, PPV (75% vs. 36.8%, NPV (100% vs. 95.8%, and accuracy (95.5% vs. 79.1%. Conclusions The point-of-care immunoassay test provides improved diagnostic accuracy for the detection of ROM compared to fern testing. It has the potential of improving patient management decisions, thereby minimizing serious complications and perinatal morbidity.

A high-performance flexible fibre-shaped electrochemical capacitor based on electrochemically reduced graphene oxide.

Science.gov (United States)

Li, Yingru; Sheng, Kaixuan; Yuan, Wenjing; Shi, Gaoquan

2013-01-11

A fibre-shaped solid electrochemical capacitor based on electrochemically reduced graphene oxide has been fabricated, exhibiting high specific capacitance and rate capability, long cycling life and attractive flexibility.

Evaluation and comparison of radio-, fluorescence, and enzyme-linked immunoassays for serum thyroxine

International Nuclear Information System (INIS)

Kaplan, L.A.; Gau, N.; Fearn, J.; Steain, E.A.; Chen, I.W.; Maxon, H.; Volle, C.

1981-01-01

We have compared three analytical systems for the measurement of serum thyroxine: enzyme-linked immunoassay (EIA), fluorescent immunoassay (FIA) and a radioimmunoassay (RIA). These were evaluated with respect to their precision, accuracy, analytical sensitivity and sample throughput. The RIA is more sensitive than the EIA (10 μg/L vs. 35 μg/L). Both systems have excellent precision (X=86 μg/L, C.V.sub(RIA)=C.V.sub(EIA)=4.6 percent). Both the EIA and RIA demonstrate good accuracy with recovery of between 97-98 percent of added thyroxine. The FIA has an apparent sensitiviity between that of the RIA and EIA (25 μg/L), but a precision consistently lower than the other two systems (C.V. =7.4 percent, X=86 μg/L). Patients' results by RIA compared well with those from EIA (r=0.91,P 0.05). Although not fully automated, the EIA performed on the Abbott ABA-100 analyzer has a sample throughput equal to the automated RIA system (Micromedic, Concept 4)

Electrochemical biotin detection based on magnetic beads and a new magnetic flow cell for screen printed electrode.

Science.gov (United States)

Biscay, Julien; González García, María Begoña; Costa García, Agustín

2015-01-01

The use of the first flow-cell for magnetic assays with an integrated magnet is reported here. The flow injection analysis system (FIA) is used for biotin determination. The reaction scheme is based on a one step competitive assay between free biotin and biotin labeled with horseradish peroxidase (B-HRP). The mixture of magnetic beads modified with streptavidin (Strep-MB), biotin and B-HRP is left 15 min under stirring and then a washing step is performed. After that, 100 μL of the mixture is injected and after 30s 100 μL of 3,3',5,5'-Tetramethylbenzidine (TMB) is injected and the FIAgram is recorded applying a potential of -0.2V. The linear range obtained is from 0.01 to 1 nM of biotin and the sensitivity is 758 nA/nM. The modification and cleaning of the electrode are performed in an easy way due to the internal magnet of the flow cell. Copyright © 2014 Elsevier B.V. All rights reserved.

Multi-site, multi-country evaluation of analytical and operational performance of a low-mid volume chemiluminescent immunoassay analyzer.

Science.gov (United States)

De Keijzer, M H; Perkins, S; Motta, V; Morelli, D; Cristol, J P; Dupuy, A M; Hong, Y; Watanabe, S; Waerdt, C; Grunewald, R W

2009-01-01

A new automated immunoassay low-mid volume (beta-hCG, BNP, and CK-MB) and the time required to complete workloads of 50 and 100 tests with a mixture of 75% routine tests and 25% stat tests was also evaluated. Total precision was typically market and in use for an existing high volume immunoassay system. Stat turn around times were consistent with the fixed analytical time of 15.6 minutes and met the expectations of the laboratories. Measured test throughput ranged from 47 - 54 tests per hour and demonstrated that the analyzer was fit for the intended purpose of supporting a laboratory that performs suitable for use.

Comparison between Amnisure Placental Alpha Microglobulin-1 Rapid Immunoassay and Standard Diagnostic Methods for Detection of Rupture of Membranes

Directory of Open Access Journals (Sweden)

Beng Kwang Ng

2013-01-01

Full Text Available Objective. To determine the diagnostic accuracy of placental alpha microglobulin-1 assay and standard diagnostic methods for detecting rupture of membrane. Study Design. Prospective diagnostic study, between June 2011 to November 2011 at a tertiary centre. Initial evaluation included both the standard diagnostic methods for rupture of membranes and placental alpha microglobulin-1 immunoassay. The actual rupture of membranes was diagnosed on review of the medical records after delivery (absence of membrane or a positive pad chart. Main Outcome Measures. Placental alpha microglobulin-1 immunoassay and standard diagnostic methods for diagnosis of rupture of membrane. Results. A total of 211 patients were recruited. At initial presentation, 187 patients (88.6% had ruptured membranes, while 24 patients (11.4% had intact membranes. Placental alpha microglobulin-1 immunoassay confirmed rupture of membranes at initial presentation with a sensitivity of 95.7% (179 of 187, specificity of 100% (24 of 24, positive predictive value of 100% (179 of 179, and negative predictive value of 75.0% (24 of 32. By comparison, the conventional standard diagnostic methods had a sensitivity of 78.1% (146 of 187, specificity of 100% (24 of 24, positive predictive value of 100% (146 of 146, and negative predictive value of 36.9% (24 of 65 in diagnosing rupture of membrane. Conclusion. Placental alpha-microglobulin-1 immunoassay is a rapid and accurate method for confirming the diagnosis of rupture of membrane. It was superior to conventional standard diagnostic methods (pooling, nitrazine, and ferning, the nitrazine test alone or fern test alone.

Rapid and Highly Sensitive Non-Competitive Immunoassay for Specific Detection of Nodularin

Directory of Open Access Journals (Sweden)

Sultana Akter

2017-09-01

Full Text Available Nodularin (NOD is a cyclic penta-peptide hepatotoxin mainly produced by Nodularia spumigena, reported from the brackish water bodies of various parts of the world. It can accumulate in the food chain and, for safety reasons, levels of NOD not only in water bodies but also in food matrices are of interest. Here, we report on a non-competitive immunoassay for the specific detection of NOD. A phage display technique was utilized to interrogate a synthetic antibody phage library for binders recognizing NOD bound to an anti-ADDA (3-Amino-9-methoxy-2,6,8-trimethyl-10-phenyldeca-4(E,6(E-dienoic acid monoclonal antibody (Mab. One of the obtained immunocomplex binders, designated SA32C11, showed very high specificity towards nodularin-R (NOD-R over to the tested 10 different microcystins (microcystin-LR, -dmLR, -RR, -dmRR, -YR, -LY, -LF, -LW, -LA, -WR. It was expressed in Escherichia coli as a single chain antibody fragment (scFv fusion protein and used to establish a time-resolved fluorometry-based assay in combination with the anti-ADDA Mab. The detection limit (blank + 3SD of the immunoassay, with a total assay time of 1 h 10 min, is 0.03 µg/L of NOD-R. This represents the most sensitive immunoassay method for the specific detection of NOD reported so far. The assay was tested for its performance to detect NOD using spiked (0.1 to 3 µg/L of NOD-R water samples including brackish sea and coastal water and the recovery ranged from 79 to 127%. Furthermore, a panel of environmental samples, including water from different sources, fish and other marine tissue specimens, were analyzed for NOD using the assay. The assay has potential as a rapid screening tool for the analysis of a large number of water samples for the presence of NOD. It can also find applications in the analysis of the bioaccumulation of NOD in marine organisms and in the food chain.

Electrochemical Analysis of Neurotransmitters

Science.gov (United States)

Bucher, Elizabeth S.; Wightman, R. Mark

2015-07-01

Chemical signaling through the release of neurotransmitters into the extracellular space is the primary means of communication between neurons. More than four decades ago, Ralph Adams and his colleagues realized the utility of electrochemical methods for the study of easily oxidizable neurotransmitters, such as dopamine, norepinephrine, and serotonin and their metabolites. Today, electrochemical techniques are frequently coupled to microelectrodes to enable spatially resolved recordings of rapid neurotransmitter dynamics in a variety of biological preparations spanning from single cells to the intact brain of behaving animals. In this review, we provide a basic overview of the principles underlying constant-potential amperometry and fast-scan cyclic voltammetry, the most commonly employed electrochemical techniques, and the general application of these methods to the study of neurotransmission. We thereafter discuss several recent developments in sensor design and experimental methodology that are challenging the current limitations defining the application of electrochemical methods to neurotransmitter measurements.

Increasing of sensitivity of fluorescent immunoassay analysis of alpha-fetoprotein by means of plasmonical silver nanoparticles

International Nuclear Information System (INIS)

Vashchenko, S.V.; Min'ko, A.A.; Romanenko, A.A.; Gaponenko, S.V.; Kulakovich, O.S.

2014-01-01

A test system is proposed based on metal enhanced fluorescence to analyze low concentrations of alpha-fetoprotein (AFP), a tumor marker. Antigen-antibody reaction was performed on polystyrene plates coated with silver nanoparticles to increase sensitivity of fluorescent immunoassay and signal-to-noise ratio as compared to silver-free system. As compared to widely used ELISA technique and other immunoassay techniques the proposed approach is characterized by smaller probe volume, fast analysis and simplicity. The proposed test system uses layer-by-layer assembly approach, LED excitation and nanowatt photodetection set-up. The proposed test system offers AFP detection at concentrations used in clinical practice. Fluorescence enhancement for labeled AFP antibodies on a silver substrate was found to depend on antibodies concentration and was up to 6 times. (authors)

Magnetic permeability based diagnostic test for the determination of the canine C-reactive protein concentration in undiluted whole blood

International Nuclear Information System (INIS)

Ibraimi, Filiz; Kriz, Kirstin; Merin, Henrik; Kriz, Dario

2009-01-01

We describe an one-step 11-min magnetic permeability based two-site immunoassay for C-reactive protein (CRP) utilizing polyclonal anti-canine CRP antibody conjugated dextran iron oxide nanoparticles (79 nm) as superparamagnetic labels and polyclonal anti-canine CRP conjugated silica microparticles (15 to 40 μm) as carriers. An inductance based magnetic permeability reader was used to detect the target analyte, CRP, in 10 μL whole blood samples, by measuring the magnetic permeability increase of the silica microparticle sediment due to immuno complex superparamagnetic nanoparticles. Measurements on standards showed a linear response between 0 and 17.5 mg/L CRP. Measurements performed on 16 whole blood samples from mixed breeds showed good correlation with a commercially available ELISA assay.

Evaluation of Beckman Coulter DxI 800 immunoassay system using clinically oriented performance goals.

Science.gov (United States)

Akbas, Neval; Schryver, Patricia G; Algeciras-Schimnich, Alicia; Baumann, Nikola A; Block, Darci R; Budd, Jeffrey R; Gaston, S J Stephen; Klee, George G

2014-11-01

We evaluated the analytical performance of 24 immunoassays using the Beckman Coulter DxI 800 immunoassay systems at Mayo Clinic, Rochester, MN for trueness, precision, detection limits, linearity, and consistency (across instruments and reagent lots). Clinically oriented performance goals were defined using the following methods: trueness-published desirable accuracy limits, precision-published desirable biologic variation; detection limits - 0.1 percentile of patient test values, linearity - 50% of total error, and consistency-percentage test values crossing key decision points. Local data were collected for precision, linearity, and consistency. Data were provided by Beckman Coulter, Inc. for trueness and detection limits. All evaluated assays except total thyroxine were within the proposed goals for trueness. Most of the assays met the proposed goals for precision (86% of intra-assay results and 75% of inter-assay results). Five assays had more than 15% of the test results below the minimum detection limits. Carcinoembryonic antigen, total thyroxine and free triiodothyronine exceeded the proposed goals of ±6.3%, ±5% and ±5.7% for dilution linearity. All evaluated assays were within the proposed goals for instrument consistency. Lot-to-lot consistency results for cortisol, ferritin and total thyroxine exceeded the proposed goals of 3.3%, 11.4% and 7% at one medical decision level, while vitamin B12 exceeded the proposed goals of 5.2% and 3.8% at two decision levels. The Beckman Coulter DxI 800 immunoassay system meets most of these proposed goals, even though these clinically focused performance goals represent relatively stringent limits. Copyright © 2014 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Electrochemical copolymerization of thiophene derivatives; a precursor to photovoltaic devices

International Nuclear Information System (INIS)

Kumar, Prajwal; Ranjith, K.; Gupta, Satyajit; Ramamurthy, Praveen C.

2011-01-01

Highlights: → This work presents an electrochemical technique for the polymerization and copolymerization of thiophene derivatives. → Copolymerization of thiophene based monomers like 7,9-dithiophene-2yl-8H-cyclopenta[a]acenaphthalene-8-one and 3-hexylthiophene are synthesized. → Study of the structural, optical, thermal, conductivity, morphological, band gap and impedance measurements of the copolymer indicates a suitable material for photovoltaic applications. - Abstract: This work presents an electrochemical technique for the polymerization and copolymerization of thiophene derivatives like 7,9-dithiophene-2yl-8H-cyclopenta[a]acenaphthalene-8-one and 3-hexylthiophene. The structural characterization of chemically synthesized monomers and electrochemically synthesized polymers was carried out by nuclear magnetic resonance and Fourier transform infrared spectroscopy. Thermal characterizations indicate that copolymer has increased thermal stability than that of homopolymer. Morphological studies of the polymerized films carried out by scanning electron microscopy shows network structure of copolymer. Optical properties of the homopolymers and copolymer were studied by UV-visible spectrometer and it was observed that band gap of copolymer is less than the homopolymers. HOMO and LUMO levels, band gap values of the respective polymers were also calculated from the cyclic voltammetry technique with various scan rates. By the peak current obtained from various scan rates shows that all polymerization reactions are diffusion controlled process. Charge transfer resistances of polymers were determined using Nyquist plots. Conductivity of synthesized polymers shows higher conductivity for copolymer than homopolymers.

Electrochemical oxidation of organic waste

International Nuclear Information System (INIS)

Almon, A.C.; Buchanan, B.R.

1990-01-01

Both silver catalyzed and direct electrochemical oxidation of organic species are examined in analytical detail. This paper describes the mechanisms, reaction rates, products, intermediates, capabilities, limitations, and optimal reaction conditions of the electrochemical destruction of organic waste. A small bench-top electrocell being tested for the treatment of small quantities of laboratory waste is described. The 200-mL electrochemical cell used has a processing capacity of 50 mL per day, and can treat both radioactive and nonradioactive waste. In the silver catalyzed process, Ag(I) is electrochemically oxidized to Ag(II), which attacks organic species such as tributylphosphate (TBP), tetraphenylborate (TPB), and benzene. In direct electrochemical oxidation, the organic species are destroyed at the surface of the working electrode without the use of silver as an electron transfer agent. This paper focuses on the destruction of tributylphosphate (TBP), although several organic species have been destroyed using this process. The organic species are converted to carbon dioxide, water, and inorganic acids

Detection of hidden hazelnut protein in food by IgY-based indirect competitive enzyme-immunoassay

NARCIS (Netherlands)

Baumgartner, S.; Bremer, M.G.E.G.; Kemmers - Voncken, A.E.M.; Smits, N.G.E.; Haasnoot, W.; Banks, J.; Reece, P.; Danks, C.; Tomkies, V.; Immer, U.; Schmitt, K.; Krska, R.

2004-01-01

The development of an indirect competitive enzyme-immunoassay for the detection of hidden hazelnut protein in complex food matrices is described. A sensitive and selective polyclonal antibody was raised by immunisation of laying hens with protein extracts from roasted hazelnuts. In contrast to

Ervaringen met een solid phase enzyme immunoassay voor het aantonen van gonorroe bij promiscue vrouwen

NARCIS (Netherlands)

Ulsen; J.van*; Michel; M.F.*; Strik; R.van*; Joost; T.H.van*; Stolz; E.*; Eijk; R.V.W.van

1985-01-01

De Gonozyme test (Abbott Laboratories), een nieuwe enzyme immunoassay (EIA) voor het aantonen van Neisseria gonorrhoeae werd geevalueerd in een grote groep promiscue vrouwen. Als de EIA werd uitgevoerd met materiaal afkomstig van de cervix, bedroeg de prevalentie van gonorroe 8,2%. Vergeleken

Electrochemical, morphological and microstructural characterization of carbon film resistor electrodes for application in electrochemical sensors

International Nuclear Information System (INIS)

Gouveia-Caridade, Carla; Soares, David M.; Liess, Hans-Dieter; Brett, Christopher M.A.

2008-01-01

The electrochemical and microstructural properties of carbon film electrodes made from carbon film electrical resistors of 1.5, 15, 140 Ω and 2.0 kΩ nominal resistance have been investigated before and after electrochemical pre-treatment at +0.9 V vs SCE, in order to assess the potential use of these carbon film electrodes as electrochemical sensors and as substrates for sensors and biosensors. The results obtained are compared with those at electrodes made from previously investigated 2 Ω carbon film resistors. Cyclic voltammetry was performed in acetate buffer and phosphate buffer saline electrolytes and the kinetic parameters of the model redox system Fe(CN) 6 3-/4- obtained. The 1.5 Ω resistor electrodes show the best properties for sensor development with wide potential windows, similar electrochemical behaviour to those of 2 Ω and close-to-reversible kinetic parameters after electrochemical pre-treatment. The 15 and 140 Ω resistor electrodes show wide potential windows although with slower kinetics, whereas the 2.0 kΩ resistor electrodes show poor cyclic voltammetric profiles even after pre-treatment. Electrochemical impedance spectroscopy related these findings to the interfacial properties of the electrodes. Microstructural and morphological studies were carried out using contact mode Atomic Force Microscopy (AFM), Confocal Raman spectroscopy and X-ray diffraction. AFM showed more homogeneity of the films with lower nominal resistances, related to better electrochemical characteristics. X-ray diffraction and Confocal Raman spectroscopy indicate the existence of a graphitic structure in the carbon films

Development of a lab-on-chip electrochemical immunosensor for detection of Polycyclic Aromatic Hydrocarbons (PAH) in environmental water

Science.gov (United States)

Felemban, Shifa; Vazquez, Patricia; Dehnert, Jan; Goridko, Vadim; Tijero, Maria; Moore, Eric

2017-06-01

The work described in this manuscript focuses on how the integration of immunoassay techniques in combination with electrochemical detection can provide a portable and very accurate solution for detection of water pollutants that are detrimental for human health. In particular, we focus our work on the quantification of polycyclic aromatic hydrocarbons (PAHs) in polluted water. Our integrative approach facilitates a real-time detection of this family of organic compounds, by reducing the time of analysis to less than one hour. Additionally, the use of a lab-on-a-chip platform delivers a portable solution that could be used in situ. Optimization of a displacement assay that investigates the presence and concentration of Benzo[a]pyrene in water, allows with the miniaturization of the standard ELISA format into a highly accurate system that provides fast results. The limits of detection obtained are comparable to those of available state-of-the art tools, and achieve the values set by European Drinking Water Directive, 0.10ng/l, as the limit for PAHs in drinking water.

340nm UV LED excitation in time-resolved fluorescence system for europium-based immunoassays detection

DEFF Research Database (Denmark)

Rodenko, Olga; Fodgaard, Henrik; Tidemand-Lichtenberg, Peter

2017-01-01

In immunoassay analyzers for in-vitro diagnostics, Xenon flash lamps have been widely used as excitation light sources. Recent advancements in UV LED technology and its advantages over the flash lamps such as smaller footprint, better wall-plug efficiency, narrow emission spectrum......, and no significant afterglow, have made them attractive light sources for gated detection systems. In this paper, we report on the implementation of a 340 nm UV LED based time-resolved fluorescence system based on europium chelate as a fluorescent marker. The system performance was tested with the immunoassay based...... on the cardiac marker, TnI. The same signal-to-noise ratio as for the flash lamp based system was obtained, operating the LED below specified maximum current. The background counts of the system and its main contributors were measured and analyzed. The background of the system of the LED based unit was improved...

Morphological reason for enhancement of electrochemical double layer capacitances of various acetylene blacks by electrochemical polarization

International Nuclear Information System (INIS)

Kim, Taegon; Ham, Chulho; Rhee, Choong Kyun; Yoon, Seong-Ho; Tsuji, Masaharu; Mochida, Isao

2008-01-01

Enhancement of electrochemical capacitance and morphological variations of various acetylene blacks caused by electrochemical polarization are presented. Acetylene blacks of different mean particle diameters were modified by air-oxidation and heat treatment to diversify the morphologies of the acetylene blacks before electrochemical polarization. The various acetylene blacks were electrochemically oxidized at 1.6 V (vs. Ag/AgCl) for 10 s and the polarization step was repeated until the capacitance values did not change any longer. These polarization steps enhanced the capacitances of the acetylene blacks and the specific enhancement factors range from 2 to 5.5. Such an enhancement is strongly related to morphological modification as revealed by transmission electron microscopic observations. The electrochemical polarization resulted in formation of tiny graphene sheets on the wide graphitic carbon surfaces, which were most responsible for the observed capacitive enhancement. Although the pseudo-capacitance increased after polarization by forming oxygenated species on the surfaces, its contribution to the total capacitance was less than 10%. The mechanism of the formation of the tiny graphene sheets during the electrochemical oxidation is described schematically

Rapid and reagent-saving immunoassay using innovative stirring actions of magnetic beads in microreactors in the sequential injection mode.

Science.gov (United States)

Tanaka, K; Imagawa, H

2005-12-15

We developed new ELISA techniques in sequential injection analysis (SIA) mode using microreactors with content of a few microliters. We immobilized antibodies on magnetic beads 1.0mum in diameter, injected the beads into microreactors and applied rotating magnetic fields of several hundred gauss. Magnetic beads, suspended in liquid in density of approximately 10(9)-10(10) particles per millilitre, form a large number of thin rod clusters, whose length-wise axes are oriented in parallel with the magnetic field. We rotate the Nd magnets below the center of the microreactor by a tiny motor at about 2000-5000rpm. These rotating clusters remarkably accelerate the binding rate of the antibodies with antigens in the liquid. The beads are trapped around the center of the rotating magnetic field even in the flowing liquid. This newly found phenomenon enables easy bead handling in microreactors. Modification of reactor walls with selected blocking reagents was essential, because protein-coated beads often stick to the wall surface and cannot move freely. Washing steps were also shortened.

Review on the progress in synthesis and application of magnetic carbon nanocomposites

Science.gov (United States)

Zhu, Maiyong; Diao, Guowang

2011-07-01

This review focuses on the synthesis and application of nanostructured composites containing magnetic nanostructures and carbon-based materials. Great progress in fabrication of magnetic carbon nanocomposites has been made by developing methods including filling process, template-based synthesis, chemical vapor deposition, hydrothermal/solvothermal method, pyrolysis procedure, sol-gel process, detonation induced reaction, self-assembly method, etc. The applications of magnetic carbon nanocomposites expanded to a wide range of fields such as environmental treatment, microwave absorption, magnetic recording media, electrochemical sensor, catalysis, separation/recognization of biomolecules and drug delivery are discussed. Finally, some future trends and perspectives in this research area are outlined.

Investigating Non-Equilibrium Fluctuations of Nanocolloids in a Magnetic Field Using Direct Imaging Methods

Science.gov (United States)

Rice, Ashley; Oprisan, Ana; Oprisan, Sorinel; Rice-Oprisan College of Charleston Team

Nanoparticles of iron oxide have a high surface area and can be controlled by an external magnetic field. Since they have a fast response to the applied magnetic field, these systems have been used for numerous in vivo applications, such as MRI contrast enhancement, tissue repair, immunoassay, detoxification of biological fluids, hyperthermia, drug delivery, and cell separation. We performed three direct imaging experiments in order to investigate the concentration-driven fluctuations using magnetic nanoparticles in the absence and in the presence of magnetic field. Our direct imaging experimental setup involved a glass cell filled with magnetic nanocolloidal suspension and water with the concentration gradient oriented against the gravitational field and a superluminescent diode (SLD) as the light source. Nonequilibrium concentration-driven fluctuations were recorded using a direct imaging technique. We used a dynamic structure factor algorithm for image processing in order to compute the structure factor and to find the power law exponents. We saw evidence of large concentration fluctuations and permanent magnetism. Further research will use the correlation time to approximate the diffusion coefficient for the free diffusion experiment. Funded by College of Charleston Department of Undergraduate Research and Creative Activities SURF grant.

Variance function estimation for immunoassays

International Nuclear Information System (INIS)

Raab, G.M.; Thompson, R.; McKenzie, I.

1980-01-01

A computer program is described which implements a recently described, modified likelihood method of determining an appropriate weighting function to use when fitting immunoassay dose-response curves. The relationship between the variance of the response and its mean value is assumed to have an exponential form, and the best fit to this model is determined from the within-set variability of many small sets of repeated measurements. The program estimates the parameter of the exponential function with its estimated standard error, and tests the fit of the experimental data to the proposed model. Output options include a list of the actual and fitted standard deviation of the set of responses, a plot of actual and fitted standard deviation against the mean response, and an ordered list of the 10 sets of data with the largest ratios of actual to fitted standard deviation. The program has been designed for a laboratory user without computing or statistical expertise. The test-of-fit has proved valuable for identifying outlying responses, which may be excluded from further analysis by being set to negative values in the input file. (Auth.)

Europium(III) chelate-dyed nanoparticles as donors in a homogeneous proximity-based immunoassay for estradiol

International Nuclear Information System (INIS)

Kokko, Leena; Sandberg, Kaisa; Loevgren, Timo; Soukka, Tero

2004-01-01

Nanoparticles containing thousands of fluorescent europium(III) chelates have a very high specific activity compared to traditional lanthanide chelate labels. It can be assumed that if these particles are used in a homogeneous assay as donors, multiple chelates can excite a single acceptor in turns and the energy transfer to the acceptor is increased. The principle was employed in an immunoassay using luminescent resonance energy transfer from a long lifetime europium(III) chelate-dyed nanoparticle to a short lifetime, near-infrared fluorescent molecule. Due to energy transfer fluorescence lifetime of the sensitised emission was prolonged and fluorescence could be measured using a time-resolved detection. A competitive homogeneous immunoassay for estradiol was created using 92 nm europium(III) chelate-dyed nanoparticle coated with 17β-estradiol specific recombinant antibody Fab fragments as a donor and estradiol conjugated with near-infrared dye AlexaFluor 680 as an acceptor. The density of Fab fragments on the surface of the particle influenced the sensitivity of the immunoassay. The optimal Fab density was reached when the entire surface of the particle participated in the energy transfer, but the areas where the energy was transferred to a single acceptor, did not overlap. We were able to detect estradiol concentrations down to 70 pmol l -1 (3xSD of a standard containing 0 nmol l -1 of E2) using a 96-well platform. In this study we demonstrated that nanoparticles containing lanthanide chelates could be used as efficient donors in homogeneous assays

Corrosion of NdFeB permanent magnet materials

International Nuclear Information System (INIS)

Warren, G.W.; Gao, G.; Li, Q.

1991-01-01

NdFeB is an important class of new magnetic materials, however corrosion resistance is a serious concern and literature on the electrochemical behavior of NdFeB is scarce. This paper reports the results of an electrochemical investigation of the corrosion behavior of sintered NdFeB magnets obtained from three manufacturers. Linear polarization (cyclic voltammetry) experiments were conducted in aqueous solutions ranging in pH from 0.7 to 13.5. A limited degree of passivation was observed in all solutions which is believed to be due to the formation of a complex Fe-Nd oxide and/or hydroxide film. The presence of a small amount of chloride ion, 10 to 100 ppm, shows only a slight effect, but higher concentrations (1000 ppm) cause a total breakdown in passivity and a dramatic increase in anodic current. The cathodic potential sweep shows an abrupt and unusual oxidation process, giving rise to an oxidation peak not commonly seen. This peak may result from dissolution of the film or preferential attack of intergranular phases

Positive predictive value of the immunoassay for Clostridium difficile toxin A and B detection at a private hospital.

Science.gov (United States)

Pérez-Topete, S E; Miranda-Aquino, T; Hernández-Portales, J A

Clostridium difficile (C. difficile) is a Gram-positive bacillus that is a common cause of diarrhea in the hospital environment, with a documented incidence of up to 10%. There are different methods to detect it, but a widely used test in our environment is the immunoassay for toxins A and B. The aim of our study was to 1) estimate the positive predictive value of the immunoassay for the detection of the C. difficile toxins A and B, 2) to establish the incidence of C. difficile-associated diarrhea in the hospital, and 3) to know the most common associated factors. A diagnostic test accuracy study was conducted within the time frame of January 2010 to August 2013 at the Hospital Christus Muguerza® Alta Especialidad on patients with symptoms suggestive of C. difficile-associated diarrhea that had a positive immunoassay test and confirmation of C. difficile through colon biopsy and stool culture. The immunoassay for toxins A and B was performed in 360 patients. Fifty-five of the cases had positive results, 35 of which showed the presence of C. difficile. Incidence was 10.2% and the positive predictive value of the test for C. difficile toxins A and B was 0.64 (95% CI, 0.51-0.76). Previous antibiotic therapy (n=29) and proton pump inhibitor use (n=19) were the most common associated factors. C. difficile incidence in our environment is similar to that found in the literature reviewed, but the positive predictive value of the test for toxin A and B detection was low. Copyright © 2016 Asociación Mexicana de Gastroenterología. Publicado por Masson Doyma México S.A. All rights reserved.

Efficient capture of magnetic microbeads by sequentially switched electroosmotic flow—an experimental study

International Nuclear Information System (INIS)

Das, Debarun; Al-Rjoub, Marwan F; Banerjee, Rupak K; Heineman, William R

2016-01-01

Magnetophoretic separation is a commonly used immunoassay technique in microfluidic platforms where magnetic microbeads (mMBs) coated with specific epitopes (antibodies) entrap target pathogens by antigen-antibody kinetics. The mMB-cell complexes are then separated from the continuous flow using an external magnetic field. The goal of this study was to design and test a microfluidic device for efficient separation of fluorescence-tagged mMBs driven by electroosmotic flow (EOF) under steady (time invariant) and switched (time varying) electric field conditions. The EOF was driven at electric fields of 100–180 V cm −1 . The mMBs were captured by a neodymium (NdFeB) permanent earth magnet. The capture efficiency ( η c ) of these mMBs was improved by sequential switching of the applied electric field driven-EOF. The fluorescent images of the captured mMBs, obtained using an inverted epifluorescence microscope, were quantified using image processing tools. In steady EOF, induced by constant electric field, the number of captured mMBs decreased by 72.3% when the electric field was increased from 100 V cm −1 to 180 V cm −1 . However, alternating the direction of flow through sequential switching of EOF increased the η c by bringing the escaped mMBs back to the capture zone and increasing their residence time in the area of higher magnetic fields. The average increase in η c was 54.3% for an mMB concentration of 1  ×  10 6 beads ml −1 ( C 1 ) and 41.6% for a concentration of 2  ×  10 6 beads ml −1 ( C 2 ). These improvements were particularly significant at higher electric fields where the η c with switching was, on average, ∼70% more compared to flow without switching. The technique of sequential switching demonstrates an efficient method for capture of mMBs for application in magnetophoretic immunoassay. (paper)

Efficient capture of magnetic microbeads by sequentially switched electroosmotic flow—an experimental study

Science.gov (United States)

Das, Debarun; Al-Rjoub, Marwan F.; Heineman, William R.; Banerjee, Rupak K.

2016-05-01

Magnetophoretic separation is a commonly used immunoassay technique in microfluidic platforms where magnetic microbeads (mMBs) coated with specific epitopes (antibodies) entrap target pathogens by antigen-antibody kinetics. The mMB-cell complexes are then separated from the continuous flow using an external magnetic field. The goal of this study was to design and test a microfluidic device for efficient separation of fluorescence-tagged mMBs driven by electroosmotic flow (EOF) under steady (time invariant) and switched (time varying) electric field conditions. The EOF was driven at electric fields of 100-180 V cm-1. The mMBs were captured by a neodymium (NdFeB) permanent earth magnet. The capture efficiency (η c) of these mMBs was improved by sequential switching of the applied electric field driven-EOF. The fluorescent images of the captured mMBs, obtained using an inverted epifluorescence microscope, were quantified using image processing tools. In steady EOF, induced by constant electric field, the number of captured mMBs decreased by 72.3% when the electric field was increased from 100 V cm-1 to 180 V cm-1. However, alternating the direction of flow through sequential switching of EOF increased the η c by bringing the escaped mMBs back to the capture zone and increasing their residence time in the area of higher magnetic fields. The average increase in η c was 54.3% for an mMB concentration of 1  ×  106 beads ml-1 (C 1) and 41.6% for a concentration of 2  ×  106 beads ml-1 (C 2). These improvements were particularly significant at higher electric fields where the η c with switching was, on average, ~70% more compared to flow without switching. The technique of sequential switching demonstrates an efficient method for capture of mMBs for application in magnetophoretic immunoassay.

Electrochemical gating in scanning electrochemical microscopy

NARCIS (Netherlands)

Ahonen, P.; Ruiz, V.; Kontturi, K.; Liljeroth, P.; Quinn, B.M.

2008-01-01

We demonstrate that scanning electrochemical microscopy (SECM) can be used to determine the conductivity of nanoparticle assemblies as a function of assembly potential. In contrast to conventional electron transport measurements, this method is unique in that electrical connection to the film is not

On-Chip Immunoassay for Determination of Urinary Albumin

Directory of Open Access Journals (Sweden)

Adisorn Tuantranont

2009-12-01

Full Text Available An immunoassay performed on a portable microfluidic device was evaluated for the determination of urinary albumin. An increase in absorbance at 500 nm resulting from immunoagglutination was monitored directly on the poly(dimethylsiloxane (PDMS microchip using a portable miniature fibre-optic spectrometer. A calibration curve was linear up to 10 mg L–1 (r2 = 0.993, with a detection limit of 0.81 mg L–1 (S/N = 3. The proposed system showed good precision, with relative standard deviations (RSDs of 5.1%, when evaluated with 10 mg L–1 albumin (n = 10. Determination of urinary albumin with the proposed system gave results highly similar to those determined by the conventional spectrophotometric method using immunoturbidimetric detection (r2 = 0.995; n = 15.

Electrochemical Immunosensor for the Detection of Aflatoxin B₁ in Palm Kernel Cake and Feed Samples.

Science.gov (United States)

Azri, Farah Asilah; Selamat, Jinap; Sukor, Rashidah

2017-11-30

Palm kernel cake (PKC) is the solid residue following oil extraction of palm kernels and useful to fatten animals either as a single feed with only minerals and vitamins supplementation, or mixed with other feedstuffs such as corn kernels or soy beans. The occurrence of mycotoxins (aflatoxins, ochratoxins, zearalenone, and fumonisins) in feed samples affects the animal's health and also serves as a secondary contamination to humans via consumption of eggs, milk and meats. Of these, aflatoxin B₁ (AFB₁) is the most toxically potent and a confirmed carcinogen to both humans and animals. Methods such as High Performance Liquid Chromatography (HPLC) and Liquid Chromatography-Mass Spectrometry (LC-MS/MS) are common in the determination of mycotoxins. However, these methods usually require sample pre-treatment, extensive cleanup and skilled operator. Therefore, in the present work, a rapid method of electrochemical immunosensor for the detection of AFB₁ was developed based on an indirect competitive enzyme-linked immunosorbent assay (ELISA). Multi-walled carbon nanotubes (MWCNT) and chitosan (CS) were used as the electrode modifier for signal enhancement. N -ethyl- N '-(3-dimethylaminopropyl)-carbodiimide (EDC) and N -hydroxysuccinimide (NHS) activated the carboxyl groups at the surface of nanocomposite for the attachment of AFB₁-BSA antigen by covalent bonding. An indirect competitive reaction occurred between AFB₁-BSA and free AFB₁ for the binding site of a fixed amount of anti-AFB₁ antibody. A catalytic signal based on horseradish peroxidase (HRP) in the presence of hydrogen peroxide (H₂O₂) and 3,3',5,5'-tetramethylbenzidine (TMB) mediator was observed as a result of attachment of the secondary antibody to the immunoassay system. As a result, the reduction peak of TMB (Ox) was measured by using differential pulse voltammetry (DPV) analysis. Based on the results, the electrochemical surface area was increased from 0.396 cm² to 1.298 cm² due to the

Nuclear Magnetic Resonance Imaging of Li-ion Battery

Directory of Open Access Journals (Sweden)

D. Ohno

2010-12-01

Full Text Available Nuclear magnetic resonance (NMR imaging has high sensitivity to proton (1H and lithium (7Li. It is a useful measurement for electrolyte in Li-ion battery. 1H NMR images of lithium ion battery which is composed of LiMn2O4 / LiClO4 + propylene carbonate (PC / Li-metal have been studied. 1H NMR images of electrolyte near cathode material (LiMn2O4 showed anomalous intensity distribution, which was quite inhomogeneous. From NMR images as a function of repetition time (TR, it was concluded that the anomalous intensity distribution was not due to change of relaxation time but an indirect (spatial para-magnetization effect from cathode material. The paramagnetization induced by high magnetic field distorts linearity of magnetic gradient field, leading to apparent intensity variance. This functional image is an easy diagnostic measurement for magnetization of cathode material, which allows the possibility to check uniformity of cathode material and change of magnetization under electrochemical process.

Rapid screening of aflatoxin B1 in beer by fluorescence polarization immunoassay.

Science.gov (United States)

Beloglazova, N V; Eremin, S A

2015-09-01

This manuscript describes the development of a sensitive, fast and easily-performed fluorescence polarization immunoassay (FPIA) for the mycotoxin aflatoxin B1 (AFB1) in various beer samples, both lager and dark. The highest sensitivity was determined for six poly- and monoclonal antibodies selective towards aflatoxins. The sample pretreatment design was emphasized since beer samples are characterized by extremely diverse matrices. Herein, the choice of sorbent for effective removal of matrix interferences prior to analysis was crucial. The samples were diluted with a borate buffer solution containing 1% PEG 6000 and passed through the clean-up column packed with NH2-derivated silica. This sample pretreatment technique was perfectly suitable for the FPIA of lager beer samples, but for dark beer and ale it did not suffice. An artificial matrix was constructed to plot a calibration curve and quantify the results of the latter samples. The developed immunoassay was characterized by a limit of detection of 1 ng mL(-1). Apparent recovery values of 89-114% for lager and 80-125% for dark beer were established. The FPIA data for AFB1 was characterized by elevated linear regression coefficients, 0.9953 for spiked lager and 0.9895 for dark beer samples respectively. Copyright © 2015 Elsevier B.V. All rights reserved.

Multiplex immunoassays for quantification of cytokines, growth factors, and other proteins in stem cell communication

Czech Academy of Sciences Publication Activity Database

Valeková, Ivona; Kupcová Skalníková, Helena; Jarkovská, Karla; Motlík, Jan; Kovářová, Hana

-, č. 1212 (2015), s. 39-63 ISSN 1940 -6029 R&D Projects: GA MŠk ED2.1.00/03.0124; GA TA ČR(CZ) TA01011466 Institutional support: RVO:67985904 Keywords : multiplex immunoassays * antibody microarray * luminex xMAP Subject RIV: EB - Genetics ; Molecular Biology

A facile and flexible process of β-cyclodextrin grafted on Fe3O4 magnetic nanoparticles and host-guest inclusion studies

International Nuclear Information System (INIS)

Zhu Jie; He Jiang; Du Xiaoyan; Lu Ruihua; Huang Lizhen; Ge Xia

2011-01-01

In this study, a kind of novel surface-functionalized magnetic nanoparticles was fabricated by the Fe 3 O 4 nanoparticles surface modification with mono-6-deoxy-6-(p-tolylsulfonyl)-cyclodextrin (6-TsO-β-CD), which were employed to interact with uric acid and their behavior was investigated by electrochemical methods. The architecture has been characterized by powder X-ray diffraction (XRD), Fourier transform infrared spectra (FT-IR), transmission electron microscopy (TEM), and thermogravimetric analysis (TGA), which confirmed that cyclodextrins have been effectively functionalized on the surface of Fe 3 O 4 nanoparticles. The analyses of vibration sample magnetometer (VSM) verified that the nanoparticles owned good magnetic property. The grafted β-cyclodextrin on the Fe3O4 nanoparticles contributed to as a modified electrode for detecting uric acid with cyclic voltammograms. Electrochemical results revealed that the new materials could exhibit excellent molecules recognition ability and show high electrochemical response. The new nanoparticles simultaneously had unique properties of magnetic nanoparticles and cyclodextrins through combining their individual distinct advantages.

Structural basis for the high specificity of a Trypanosoma congolense immunoassay targeting glycosomal aldolase.

Directory of Open Access Journals (Sweden)

Joar Pinto

2017-09-01

Full Text Available Animal African trypanosomosis (AAT is a neglected tropical disease which imposes a heavy burden on the livestock industry in Sub-Saharan Africa. Its causative agents are Trypanosoma parasites, with T. congolense and T. vivax being responsible for the majority of the cases. Recently, we identified a Nanobody (Nb474 that was employed to develop a homologous sandwich ELISA targeting T. congolense fructose-1,6-bisphosphate aldolase (TcoALD. Despite the high sequence identity between trypanosomatid aldolases, the Nb474-based immunoassay is highly specific for T. congolense detection. The results presented in this paper yield insights into the molecular principles underlying the assay's high specificity.The structure of the Nb474-TcoALD complex was determined via X-ray crystallography. Together with analytical gel filtration, the structure reveals that a single TcoALD tetramer contains four binding sites for Nb474. Through a comparison with the crystal structures of two other trypanosomatid aldolases, TcoALD residues Ala77 and Leu106 were identified as hot spots for specificity. Via ELISA and surface plasmon resonance (SPR, we demonstrate that mutation of these residues does not abolish TcoALD recognition by Nb474, but does lead to a lack of detection in the Nb474-based homologous sandwich immunoassay.The results show that the high specificity of the Nb474-based immunoassay is not determined by the initial recognition event between Nb474 and TcoALD, but rather by its homologous sandwich design. This (i provides insights into the optimal set-up of the assay, (ii may be of great significance for field applications as it could explain the potential detection escape of certain T. congolense strains, and (iii may be of general interest to those developing similar assays.

Evaluation of the Lumipulse G TP-N Chemiluminescent Immunoassay as a Syphilis Screening Test.

Science.gov (United States)

Ortiz, Daniel A; Loeffelholz, Michael J

2017-11-01

A syphilis diagnosis is often aided by the detection of treponemal and nontreponemal antibodies. Automated treponemal antibody detection systems enable high-volume clinical laboratories to perform syphilis screening at a faster pace with lower labor costs. The Lumipulse G TP-N chemiluminescent immunoassay is an automated system that qualitatively detects IgG and IgM antibodies against Treponema pallidum antigens in human serum and plasma. To assess performance characteristics and workflow efficiency, the Lumipulse G TP-N assay was compared to the Bioplex 2200 Syphilis IgG multiplex flow immunoassay. Among the 4,134 routine and HIV samples tested by the two automated assays, the percentage of agreement was excellent at 99.0% (95% confidence interval [CI], 98.6% to 99.2%; κ, 0.89), with the Lumipulse G TP-N having a shorter time to first and subsequent results. All specimens with reactive syphilis screening results were further tested by rapid plasma reagin (RPR) and Treponema pallidum particle agglutination (TP·PA) testing ( n = 231). The results from the RPR-reactive samples ( n = 82) showed complete concordance with the two automated assays, while the TP·PA assay displayed some discrepancies. The positive percent agreement (PPA) and negative percent agreement (NPA) between the TP·PA test and the Lumipulse G TP-N test were 98.9% and 77.3%, respectively. The Bioplex 2200 Syphilis IgG immunoassay displayed a similar PPA (100%) but a substantially lower NPA (15.9%). Patient chart reviews of discrepant results suggested that the Lumipulse G TP-N assay produced 27 fewer falsely reactive results and can reduce the amount of additional confirmatory RPR and TP·PA testing needed. The analogous performance characteristics of the two automated systems indicate that the Lumipulse G TP-N assay is suitable for high-throughput syphilis screening. Copyright © 2017 American Society for Microbiology.

Myeloma-Derived Light Chain Paired with a Diagnostic Monoclonal Antibody Hinders Immunoassay Performance.

Science.gov (United States)

Tu, Bailin; Tieman, Bryan; Moore, Jeffrey; Pan, You; Muerhoff, A Scott

2017-06-01

Monoclonal antibodies are widely used as the capture and detection reagents in diagnostic immunoassays. In the past, myeloma fusion partners expressing endogenous heavy and/or light chains were often used to generate hybridoma cell lines. As a result, mixed populations of antibodies were produced that can cause inaccurate test results, poor antibody stability, and significant lot-to-lot variability. We describe one such scenario where the P3U1 (P3X63Ag8U.1) myeloma fusion partner was used in the generation of a hybridoma producing protein induced vitamin K absence/antagonist-II (PIVKA II) antibody. The hybridoma produces three subpopulations of immunoglobulin as determined by ion exchange (IEx) chromatography that exhibit varying degrees of immunoreactivity (0%, 50%, or 100%) to the target antigen as determined by Surface Plasmon Resonance. To produce an antibody with the highest possible sensitivity and specificity, the antigen-specific heavy and light chain variable domains (VH and VL) were cloned from the hybridoma and tethered to murine IgG1 and kappa scaffolds. The resulting recombinant antibody was expressed in Chinese hamster ovary cells and is compatible for use in a diagnostic immunoassay.

[Membrane-filtration immunoassay: reagents, methods and the diagnostic and technical means for detection].

Science.gov (United States)

Khramov, E N; Osin, N S; Pomelova, V G; Vikha, I V; Bychenkova, T A; Smirnova, V G; Grakina, G I; Kas'ianova, T A

1999-01-01

The comprehensive development of dot-EIA made at the State Research Institute of Biological Instrument-Making Industry has provided devices KIMF-02 and KIMF-03), a base of chemical reagents, immunoassays, test systems for detection of a wide range of causative agents of viral and bacterial infections, that of serodiagnosis of their related diseases. The KIMF-02 kit has undergone engineering and medical tests and recommended for the Ministry of Health of the Russian Federation to produce them in stock. The kit includes all required for analysis even in an ill-equipped laboratory, a set of attached agents ensures a valid visual recording of results. The developed procedures and test systems allow the immunoassay to be as sensitive as TIFA; however, they are laborious and much simpler in design. The simple and rapid procedures of dot-EIA are recommended for incorporation into the a package of laboratory methods for verification of the accumulation of virus-specific antigens in various biological substrata, environmental samples, for control of the activity of antigens and antibodies used in serological tests, for detection of specific antigens in the clinical samples, and for serodiagnosis of infections.

Thermophilic Campylobacter spp. in turkey samples: evaluation of two automated enzyme immunoassays and conventional microbiological techniques

DEFF Research Database (Denmark)

Borck, Birgitte; Stryhn, H.; Ersboll, A.K.

2002-01-01

Aims: To determine the sensitivity and specificity of two automated enzyme immunoassays (EIA), EiaFoss and Minividas, and a conventional microbiological culture technique for detecting thermophilic Campylobacter spp. in turkey samples. Methods and Results: A total of 286 samples (faecal, meat...

Kinetic mechanism for modeling of electrochemical reactions.

Science.gov (United States)

Cervenka, Petr; Hrdlička, Jiří; Přibyl, Michal; Snita, Dalimil

2012-04-01

We propose a kinetic mechanism of electrochemical interactions. We assume fast formation and recombination of electron donors D- and acceptors A+ on electrode surfaces. These mediators are continuously formed in the electrode matter by thermal fluctuations. The mediators D- and A+, chemically equivalent to the electrode metal, enter electrochemical interactions on the electrode surfaces. Electrochemical dynamics and current-voltage characteristics of a selected electrochemical system are studied. Our results are in good qualitative agreement with those given by the classical Butler-Volmer kinetics. The proposed model can be used to study fast electrochemical processes in microsystems and nanosystems that are often out of the thermal equilibrium. Moreover, the kinetic mechanism operates only with the surface concentrations of chemical reactants and local electric potentials, which facilitates the study of electrochemical systems with indefinable bulk.

Effect of cerium on the corrosion behaviour of sintered (Nd,Ce)FeB magnet

Science.gov (United States)

Yang, Lijing; Bi, Mengxue; Jiang, Jianjun; Ding, Xuefeng; Zhu, Minggang; Li, Wei; Lv, Zhongshan; Song, Zhenlun

2017-06-01

For the balanced consumption of rare-earth elements, cerium (Ce) was partially used for NdFeB magnets instead of Nd. The corrosion behaviour of the (Nd,Ce)FeB magnet with different Ce contents in 3.5% NaCl solution was investigated by SEM, XRD, EDS and electrochemical tests. After immersion, the weight loss was calculated and the magnetic properties of the samples were measured. Results showed that Ce affected the corrosion of the (Nd,Ce)FeB magnet. Compared with the NdFeB magnet without Ce but of the same grade as the magnetic energy product, (Nd,Ce)FeB magnet showed better corrosion resistance. With increased Ce content, the corrosion resistances and magnetic properties of (Nd,Ce)FeB magnets were investigated.

Competitive enzyme immunoassay for human chorionic somatomammotropin using the avidin-biotin system

International Nuclear Information System (INIS)

Rappuoli, R.; Leoncini, P.; Tarli, P.; Neri, P.

1981-01-01

Human chorionic somatomammotropin (HCS) is determined by an enzyme immunoassay where HCS competes with biotin-labeled HCS for insolubilized anti-HCS antibodies. Enzyme-labeled avidin is then used to reveal the amount of bound HCS. The system proves to be sensitive (1 ng/ml of HCS can be detected) and results agree with radioimmunoassay determinations (correlation coefficient = 0.979). Kinetics of the avidin-biotin reaction and coating of polystyrene wells are also investigated

Electrochemical magneto immunosensor based on endogenous β-galactosidase enzyme to determine enterotoxicogenic Escherichia coli F4 (K88) in swine feces using square wave voltammetry.

Science.gov (United States)

Viviana Tarditto, Lorena; Alicia Zon, María; García Ovando, Hugo; Roberto Vettorazzi, Nelio; Javier Arévalo, Fernando; Fernández, Héctor

2017-11-01

Diseases caused by enterotoxicogenic Escherichia coli F4 (K88) (ETEC F4) are a problem in swine production establishments. Due to the high rate of mortality and morbidity of E. coli infections, a rapid and accurate diagnosis is important in order to choose an appropriate treatment to reduce the economic impact. Therefore, an electrochemical magneto-immunosensor (EMI) was developed to detect and quantify ETEC F4 in swine feces samples through a direct non-competitive immunoassay. ETEC F4 was selectively captured by immunomagnetic separation. The detection principle was based on the activity of β-galactosidase endogenous enzyme (β-gal), which hydrolyses the p-aminophenyl-β-D-galactopyranoside (p-APG) producing p-aminophenol (p-AP), which was oxidized on a carbon screen printed electrode (CSPE) using square wave voltammetry (SWV). All parameters related to construction and electrochemical responses were optimized. The total analysis time to quantify ETEC F4 using the EMI was less than 2h and the limit of detection (LOD) was 33CFUmL -1 . The perceptual relative error (%E r ) was 20%. The magneto-immunosensor was validated versus conventional method of culture and plate count, obtaining a very good agreement. The EMI is simple, fast and economical to detect and quantify ETEC F4 in swine feces samples, being thus a valuable tool in swine production. Copyright © 2017 Elsevier B.V. All rights reserved.

CMOS Electrochemical Instrumentation for Biosensor Microsystems: A Review

Directory of Open Access Journals (Sweden)

Haitao Li

2016-12-01

Full Text Available Modern biosensors play a critical role in healthcare and have a quickly growing commercial market. Compared to traditional optical-based sensing, electrochemical biosensors are attractive due to superior performance in response time, cost, complexity and potential for miniaturization. To address the shortcomings of traditional benchtop electrochemical instruments, in recent years, many complementary metal oxide semiconductor (CMOS instrumentation circuits have been reported for electrochemical biosensors. This paper provides a review and analysis of CMOS electrochemical instrumentation circuits. First, important concepts in electrochemical sensing are presented from an instrumentation point of view. Then, electrochemical instrumentation circuits are organized into functional classes, and reported CMOS circuits are reviewed and analyzed to illuminate design options and performance tradeoffs. Finally, recent trends and challenges toward on-CMOS sensor integration that could enable highly miniaturized electrochemical biosensor microsystems are discussed. The information in the paper can guide next generation electrochemical sensor design.

Synthesis, characterization, phase diagrams and superconducting and normal state magnetic properties of La2-xSrxCuO4 (0 ≤ x ≤ 0.08) and electrochemically oxidized La2-xSrxCuO4+δ (0 ≤ x ≤ 0.33, 0 ≤ δ ≤ 0.12)

International Nuclear Information System (INIS)

Chou, F.

1993-08-01

La 2-x Sr x CuO 4 (0 ≤ x ≤ 0.15) can all be intercalated with oxygen by a novel electrochemical oxidation method. Bulk superconductivity is found with an onset T c ∼ 40 K for the whole range 0.01 ≤ x ≤ 0.15; for x = 0.25 and 0.33, the electrochemical oxidation did not improve the superconducting properties. The magnetic susceptibility χ(T = 50--320 K) data for La 2 CuO 4.11 and La 1.92 Sr 0.08 CuO 4.07 are nearly identical with those of conventionally prepared La 1.85 Sr 0.15 CuO 4 , indicating that the hole doping level (p) in the CuO 2 planes of the three compounds is nearly the same. Combined thermogravimetric analysis and iodometric titration experiments indicate that part of the intercalated oxygen has a formal valence close to -1. The maximum doped-hole concentration in the CuO 2 planes that can be achieved from combined Sr-doping and electrochemical oxygen doping for 0 ≤ x ≤ 0.15 is p ∼ 0.16 holes/formula unit. Oxygen can also intercalate into single crystal La 2 CuO 4 through a slow electrochemical oxidation process. The required low current and long time for the charging process reflects that the oxygen intercalation for a single crystal is limited by its small specific surface area and long diffusion distance. The anisotropic superconducting, magnetic and transport properties are summarized and compared with those of polycrystalline La 2 CuO 4+δ as well as of YBa 2 Cu 3 O 7-δ and La 2-x Sr x CuO 4 single crystals. The single crystal La 2 CuO 4+δ has a maximum T c ∼ 40 K, which is lower than that (T c ∼ 42--45) of the corresponding polycrystalline samples. The magnetic phase diagram of La 2-x Sr x CuO 4 in the antiferromagnetic (AF) regime (0 ≤ x ≤ 0.02) has been derived from 139 La NQR studies from 4 to 250 K

Lab-on-a-disc platform for screening of genetically modified E. coli cells via cell-free electrochemical detection of p-Coumaric acid

DEFF Research Database (Denmark)

Sanger, Kuldeep; Zor, Kinga; Jendresen, Christian Bille

2017-01-01

filtration and electrochemical detection units, the sample filtration was performed by rotating the disc using a programmable closed-loop stepper motor. The electrodes, patterned on plastic substrate, were connected through a printed circuit board to the slip ring using a robust magnetic clamping system...

Electrochemical immunoassay for the prostate specific antigen using a reduced graphene oxide functionalized with a high molecular-weight silk peptide

International Nuclear Information System (INIS)

Wang, Yanying; Qu, Ying; Li, Chunya; Wu, Kangbing; Liu, Guishen; Hou, Xiaodong; Huang, Yina; Wu, Wangze

2015-01-01

High molecular-weight silk peptide (SP) was used to functionalize the surface of nanosheets of reduced graphene oxide (rGO). The SP-rGO nanocomposite was then mixed with mouse anti-human prostate specific antigen monoclonal antibody (anti-PSA) and coated onto a glassy carbon electrode to fabricate an immunosensor. By using the hexacyanoferrate redox system as electroactive probe, the immunosensor was characterized by voltammetry and electrochemical impedance spectroscopy. The peak current, measured at the potential of 0.24 V (vs. SCE), is distinctly reduced after binding prostate specific antigen (PSA). Response (measured by differential pulse voltammetry) is linearly related to PSA concentration in the range from 0.1 to 5.0 ng · mL −1 and from 5.0 to 80.0 ng∙mL −1 , and the detection limit is 53 pg∙mL −1 (at an SNR of 3). The immunosensor was successfully applied to the determination of PSA in clinical serum samples, and the results were found to agree well with those obtained with an enzyme-linked immunosorbent assay. (author)

A new real-time method for investigation of affinity properties and binding kinetics of magnetic nanoparticles

Energy Technology Data Exchange (ETDEWEB)

Orlov, Alexey V. [Prokhorov General Physics Institute, Russian Academy of Sciences, 38 Vavilov St, 119991 Moscow (Russian Federation); Nikitin, Maxim P. [Prokhorov General Physics Institute, Russian Academy of Sciences, 38 Vavilov St, 119991 Moscow (Russian Federation); Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 16/10 Miklukho-Maklaya St, 117997 Moscow (Russian Federation); Moscow Institute of Physics and Technology, 9 Institutskii per., Dolgoprudny, Moscow Region 141700 (Russian Federation); Bragina, Vera A.; Znoyko, Sergey L.; Zaikina, Marina N.; Ksenevich, Tatiana I.; Gorshkov, Boris G. [Prokhorov General Physics Institute, Russian Academy of Sciences, 38 Vavilov St, 119991 Moscow (Russian Federation); Nikitin, Petr I., E-mail: nikitin@kapella.gpi.ru [Prokhorov General Physics Institute, Russian Academy of Sciences, 38 Vavilov St, 119991 Moscow (Russian Federation); National Research Nuclear University MEPhI (Moscow Engineering Physics Institute), 31 Kashirskoe shosse, 115409 Moscow (Russian Federation)

2015-04-15

A method for quantitative investigation of affinity constants of receptors immobilized on magnetic nanoparticles (MP) is developed based on spectral correlation interferometry (SCI). The SCI records with a picometer resolution the thickness changes of a layer of molecules or nanoparticles due to a biochemical reaction on a cover slip, averaged over the sensing area. The method is compatible with other types of sensing surfaces employed in biosensing. The measured values of kinetic association constants of magnetic nanoparticles are 4 orders of magnitude higher than those of molecular antibody association with antigen. The developed method also suggests highly sensitive detection of antigens in a wide dynamic range. The limit of detection of 92 pg/ml has been demonstrated for prostate-specific antigen (PSA) with 50-nm MP employed as labels, which produce 3-order amplification of the SCI signals. The calibration curve features high sensitivity (slope) of 3-fold signal raise per 10-fold increase of PSA concentration within 4-order dynamic range, which is an attractive compromise for precise quantitative and highly sensitive immunoassay. The proposed biosensing technique offers inexpensive disposable sensor chips of cover slips and represents an economically sound alternative to traditional immunoassays for disease diagnostics, detection of pathogens in food and environmental monitoring. - Highlights: • Method for study of affinity constants of magnetic nanoparticles with receptors is proposed. • Association constants of such particles are 4 orders higher than for biomolecules. • Method is compatible with widely used biosensor surfaces and affordable consumables. • It has high sensitivity: 3-fold signal increasing per 10-fold of PSA concentration. • Limit of detection for PSA is 92 pg/ml, dynamic range – 4 orders of concentration.

Electrochemical solar energy conversion

International Nuclear Information System (INIS)

Gerischer, H.

1991-01-01

The principles of solar energy conversion in photoelectrochemical cells are briefly reviewed. Cells for the generation of electric power and for energy storage in form of electrochemical energy are described. These systems are compared with solid state photovoltaic devices, and the inherent difficulties for the operation of the electrochemical systems are analyzed. (author). 28 refs, 10 figs

Concordance Between Different Amyloid Immunoassays and Visual Amyloid Positron Emission Tomographic Assessment.

Science.gov (United States)

Janelidze, Shorena; Pannee, Josef; Mikulskis, Alvydas; Chiao, Ping; Zetterberg, Henrik; Blennow, Kaj; Hansson, Oskar

2017-12-01

Visual assessment of amyloid positron emission tomographic (PET) images has been approved by regulatory authorities for clinical use. Several immunoassays have been developed to measure β-amyloid (Aβ) 42 in cerebrospinal fluid (CSF). The agreement between CSF Aβ42 measures from different immunoassays and visual PET readings may influence the use of CSF biomarkers and/or amyloid PET assessment in clinical practice and trials. To determine the concordance between CSF Aβ42 levels measured using 5 different immunoassays and visual amyloid PET analysis. The study included 262 patients with mild cognitive impairment or subjective cognitive decline from the Swedish BioFINDER (Biomarkers for Identifying Neurodegenerative Disorders Early and Reliably) cohort (recruited from September 1, 2010, through December 31, 2014) who had undergone flutemetamol F 18 ([18F]flutemetamol)-labeled PET. Levels of CSF Aβ42 were analyzed using the classic INNOTEST and the newer modified INNOTEST, fully automated Lumipulse (FL), EUROIMMUN (EI), and Meso Scale Discovery (MSD) assays. Concentrations of CSF Aβ were assessed using an antibody-independent mass spectrometry-based reference measurement procedure. The concordance of CSF Aβ42 levels and Aβ42:Aβ40 and Aβ42:tau ratios with visual [18F]flutemetamol PET status. Of 262 participants (mean [SD] age, 70.9 [5.5] years), 108 were women (41.2%) and 154 were men (58.8%). The mass spectrometry-derived Aβ42 values showed higher correlations with the modified Aβ42-INNOTEST (r = 0.97), Aβ42-FL (r = 0.93), Aβ42-EI (r = 0.93), and Aβ42-MSD (r = 0.95) assays compared with the classic Aβ42-INNOTEST assay (r = 0.88; P ≤ .01). The signal in the classic Aβ42-INNOTEST assay was partly quenched by recombinant Aβ1-40 peptide. However, the classic Aβ42-INNOTEST assay showed better concordance with visual [18F]flutemetamol PET status (area under the receiver operating characteristic curve [AUC], 0.92) compared with the

FeSi4P4: A novel negative electrode with atypical electrochemical mechanism for Li and Na-ion batteries

Science.gov (United States)

Coquil, Gaël; Fullenwarth, Julien; Grinbom, Gal; Sougrati, Moulay Tahar; Stievano, Lorenzo; Zitoun, David; Monconduit, Laure

2017-12-01

The electrochemical mechanism and performance of FeSi4P4, vs. Na and Li were studied using a combination of operando X-ray diffraction, 57Fe Mössbauer spectroscopy, and SQUID magnetometry. This silicon- and phosphorous-rich material exhibits a high capacity of 1750 mAh/g, retaining 1120 mAh/g after 40 cycles, and reacts through an original reversible mechanism surprisingly involving only slight changes in the chemical environment of the iron. Magnetic measurements and 57Fe Mössbauer spectroscopy at low temperature reveal the reversible but incomplete change of the magnetic moment upon charge and discharge. Such a mild reversible process without drastic phase transition (with the exception of the crystalline to amorphous transition during the first lithiation) can explain the satisfying capacity retention. The electrochemical mechanism appears thus to be significantly different from the classical conversion or alloying/dealloying mechanisms usually observed in Lithium ion batteries for p-group element based materials. The same iron silicon phosphide electrode shows also interesting but significantly lower performance vs. Na, with a limited capacity retention 350 mAh/g.

Characterization of Electrochemically Generated Silver

Science.gov (United States)

Adam, Niklas; Martinez, James; Carrier, Chris

2014-01-01

Silver biocide offers a potential advantage over iodine, the current state of the art in US spacecraft disinfection technology, in that silver can be safely consumed by the crew. Low concentrations of silver (Silver does not require hardware to remove it from a water system, and therefore can provide a simpler means for disinfecting water. The Russian segment of the International Space Station has utilized an electrochemically generated silver solution, which is colloidal in nature. To be able to reliably provide a silver biocide to drinking water by electrochemical means would reduce mass required for removing another biocide such as iodine from the water. This would also aid in crew time required to replace iodine removal cartridges. Future long term missions would benefit from electrochemically produced silver as the biocide could be produced on demand and requires only a small concentration to be effective. Since it can also be consumed safely, there is less mass in removal hardware and little consumables required for production. The goal of this project initially is to understand the nature of the electrochemically produced silver, the particle sizes produced by the electrochemical cell and the effect that voltage adjustment has on the particle size. In literature, it has been documented that dissolved oxygen and pH have an effect on the ionization of the electrochemical silver so those parameters would be measured and possibly adjusted to understand their effect on the silver.

Concatenation of electrochemical grafting with chemical or electrochemical modification for preparing electrodes with specific surface functionality

International Nuclear Information System (INIS)

Verma, Pallavi; Maire, Pascal; Novak, Petr

2011-01-01

Surface modified electrodes are used in electro-analysis, electro-catalysis, sensors, biomedical applications, etc. and could also be used in batteries. The properties of modified electrodes are determined by the surface functionality. Therefore, the steps involved in the surface modification of the electrodes to obtain specific functionality are of prime importance. We illustrate here bridging of two routes of surface modifications namely electrochemical grafting, and chemical or electrochemical reduction. First, by electrochemical grafting an organic moiety is covalently immobilized on the surface. Then, either by chemical or by electrochemical route the terminal functional group of the grafted moiety is transformed. Using the former route we prepared lithium alkyl carbonate (-O(CH 2 ) 3 OCO 2 Li) modified carbon with potential applications in batteries, and employing the latter we prepared phenyl hydroxyl amine (-C 6 H 4 NHOH) modified carbon which may find application in biosensors. Benzyl alcohol (-C 6 H 4 CH 2 OH) modified carbon was prepared by both chemical as well as electrochemical route. We report combinations of conjugating the two steps of surface modifications and show how the optimal route of terminal functional group modification depends on the chemical nature of the moiety attached to the surface in the electrochemical grafting step.

Concatenation of electrochemical grafting with chemical or electrochemical modification for preparing electrodes with specific surface functionality

Energy Technology Data Exchange (ETDEWEB)

Verma, Pallavi; Maire, Pascal [Paul Scherrer Institut, Electrochemistry Laboratory, Section Electrochemical Energy Storage, CH-5232 Villigen PSI (Switzerland); Novak, Petr, E-mail: petr.novak@psi.c [Paul Scherrer Institut, Electrochemistry Laboratory, Section Electrochemical Energy Storage, CH-5232 Villigen PSI (Switzerland)

2011-04-01

Surface modified electrodes are used in electro-analysis, electro-catalysis, sensors, biomedical applications, etc. and could also be used in batteries. The properties of modified electrodes are determined by the surface functionality. Therefore, the steps involved in the surface modification of the electrodes to obtain specific functionality are of prime importance. We illustrate here bridging of two routes of surface modifications namely electrochemical grafting, and chemical or electrochemical reduction. First, by electrochemical grafting an organic moiety is covalently immobilized on the surface. Then, either by chemical or by electrochemical route the terminal functional group of the grafted moiety is transformed. Using the former route we prepared lithium alkyl carbonate (-O(CH{sub 2}){sub 3}OCO{sub 2}Li) modified carbon with potential applications in batteries, and employing the latter we prepared phenyl hydroxyl amine (-C{sub 6}H{sub 4}NHOH) modified carbon which may find application in biosensors. Benzyl alcohol (-C{sub 6}H{sub 4}CH{sub 2}OH) modified carbon was prepared by both chemical as well as electrochemical route. We report combinations of conjugating the two steps of surface modifications and show how the optimal route of terminal functional group modification depends on the chemical nature of the moiety attached to the surface in the electrochemical grafting step.

Development of an ultrasensitive immunoassay for detecting tartrazine.

Science.gov (United States)

Li, Zhuokun; Song, Shanshan; Xu, Liguang; Kuang, Hua; Guo, Shidong; Xu, Chuanlai

2013-06-25

We have developed an ultrasensitive indirect competitive enzyme-linked immunosorbent assay for the determination of tartrazine. Two carboxylated analogues of tartrazine with different spacer lengths, and one derivative from commercial tartrazine after a little chemical modification, were synthesized as haptens in order to produce antibodies specific to tartrazine. The effect of sulfonic acid groups on the hapten structure of tartrazine was also studied carefully for the first time. A most specific monoclonal antibody against tartrazine was created and exhibited an IC50 value of 0.105 ng/mL and a limit of detection of 0.014 ng/mL, with no cross-reactivity to other structurally-related pigments. The established immunoassay was applied to the determination of tartrazine in fortified samples of orange juice and in real positive samples of carbonated beverages.

Electrochemical behavior of pitch-based activated carbon fibers for electrochemical capacitors

International Nuclear Information System (INIS)

Lee, Hye-Min; Kwac, Lee-Ku; An, Kay-Hyeok; Park, Soo-Jin; Kim, Byung-Joo

2016-01-01

Highlights: • Electrode materials for electrochemical capacitors were developed using pitch-based activated carbon fibers with steam activation. • Activated carbon fibers showed enhanced specific surface area from 1520 to 3230 m 2 /g. • The increase in the specific capacitance of the samples was determined by charged pore structure during charging and discharging. - Abstract: In the present study, electrode materials for electrochemical capacitors were developed using pitch-based activated carbon fibers with steam activation. The surface and structural characteristics of activated carbon fibers were observed using scanning electron microscopy and X-ray diffraction, respectively. Pore characteristics were investigated using N 2 /77 K adsorption isotherms. The activated carbon fibers were applied as electrodes for electrical double-layer capacitors and analyzed in relation to the activation time. The specific surface area and total pore volume of the activated carbon fibers were determined to be 1520–3230 m 2 /g and 0.61–1.87 cm 3 /g, respectively. In addition, when the electrochemical characteristics were analyzed, the specific capacitance was confirmed to have increased from 1.1 F/g to 22.5 F/g. From these results, it is clear that the pore characteristics of pitch-based activated carbon fibers changed considerably in relation to steam activation and charge/discharge cycle; therefore, it was possible to improve the electrochemical characteristics of the activated carbon fibers.

Bussing Structure In An Electrochemical Cell

Science.gov (United States)

Romero, Antonio L.

2001-06-12

A bussing structure for bussing current within an electrochemical cell. The bussing structure includes a first plate and a second plate, each having a central aperture therein. Current collection tabs, extending from an electrode stack in the electrochemical cell, extend through the central aperture in the first plate, and are then sandwiched between the first plate and second plate. The second plate is then connected to a terminal on the outside of the case of the electrochemical cell. Each of the first and second plates includes a second aperture which is positioned beneath a safety vent in the case of the electrochemical cell to promote turbulent flow of gasses through the vent upon its opening. The second plate also includes protrusions for spacing the bussing structure from the case, as well as plateaus for connecting the bussing structure to the terminal on the case of the electrochemical cell.

Improved supercapacitor performance of MnO2-electrospun carbon nanofibers electrodes by mT magnetic field

Science.gov (United States)

Zeng, Zheng; Liu, Yiyang; Zhang, Wendi; Chevva, Harish; Wei, Jianjun

2017-08-01

This work reports on a finding of mT magnetic field induced energy storage enhancement of MnO2-based supercapacitance electrodes (magneto-supercapacitor). Electrodes with MnO2 electrochemically deposited at electrospun carbon nanofibers (ECNFs) film are studied by cyclic voltammetry (CV), galvanostatic charge/discharge, electrochemical impedance spectroscopy (EIS), and life cycle stability tests in the presence/absence of milli-Tesla (mT) magnetic fields derived by Helmholtz coils. In the presence of a 1.34 mT magnetic field, MnO2/ECNFs shows a magneto-enhanced capacitance of 141.7 F g-1 vs. 119.2 F g-1 (∼19% increase) with absence of magnetic field at a voltage sweeping rate of 5 mV s-1. The mechanism of the magneto-supercapacitance is discussed and found that the magnetic susceptibility of the MnO2 significantly improves the electron transfer of a pseudo-redox reaction of Mn(IV)/Mn(III) at the electrode, along with the magnetic field induced impedance effect, which may greatly enhance the interface charge density, facilitate electrolyte transportation, and improve the efficiency of cation intercalation/de-intercalation of the pseudocapacitor under mT-magnetic field exposure, resulting in enhancement of energy storage capacitance and longer charge/discharge time of the MnO2/ECNFs electrode without sacrificing its life cycle stability.

Electrochemical ion separation in molten salts

Science.gov (United States)

Spoerke, Erik David; Ihlefeld, Jon; Waldrip, Karen; Wheeler, Jill S.; Brown-Shaklee, Harlan James; Small, Leo J.; Wheeler, David R.

2017-12-19

A purification method that uses ion-selective ceramics to electrochemically filter waste products from a molten salt. The electrochemical method uses ion-conducting ceramics that are selective for the molten salt cations desired in the final purified melt, and selective against any contaminant ions. The method can be integrated into a slightly modified version of the electrochemical framework currently used in pyroprocessing of nuclear wastes.

A sandwich electrochemical immunosensor for Salmonella pullorum and Salmonella gallinarum based on a screen-printed carbon electrode modified with an ionic liquid and electrodeposited gold nanoparticles

International Nuclear Information System (INIS)

Fei, Jianfeng; Dou, Wenchao; Zhao, Guangying

2015-01-01

This article describes an electrochemical immunosensor for rapid determination of Salmonella pullorum and Salmonella gallinarum. The first step in the preparation of the immunosensor involves the electrodeposition of gold nanoparticles used for capturing antibody and enhancing signals. In order to generate a benign microenvironment for the antibody, the ionic liquid (IL) 1-butyl-3-methylimidazolium hexafluorophosphate was used to modify the surface of a screen-printed carbon electrode (SPCE). The single steps of modification were monitored via cyclic voltammetry and electrochemical impedance spectroscopy. Based on these findings, a sandwich immunoassay was worked out for the two Salmonella species by immobilizing the respective unlabeled antibodies on the SPCE. Following exposure to the analytes, secondary antibody (labeled with HRP) is added to form the sandwich. After adding hydrogen peroxide and thionine, the latter is oxidized and its signal measured via CV. A linear response to the Salmonella species is obtained in the 10 4 to 10 9 cfu · mL −1 concentration range, and the detection limits are 3.0 × 10 3 cfu · mL −1 for both species (at an SNR of 3). This assay is sensitive, highly specific, acceptably accurate and reproducible. Given its low detection limit, it represents a promising tool for the detection of S. pullorum, S. gallinarum, and - conceivably - of other food-borne pathogens by exchanging the antibody. (author)

A sulfhydryl-reactive ruthenium (II complex and its conjugation to protein G as a universal reagent for fluorescent immunoassays.

Directory of Open Access Journals (Sweden)

Jing-Tang Lin

Full Text Available To develop a fluorescent ruthenium complex for biosensing, we synthesized a novel sulfhydryl-reactive compound, 4-bromophenanthroline bis-2,2'-dipyridine Ruthenium bis (hexafluorophosphate. The synthesized Ru(II complex was crosslinked with thiol-modified protein G to form a universal reagent for fluorescent immunoassays. The resulting Ru(II-protein G conjugates were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE. The emission peak wavelength of the Ru(II-protein G conjugate was 602 nm at the excitation of 452 nm which is similar to the spectra of the Ru(II complex, indicating that Ru(II-protein G conjugates still remain the same fluorescence after conjugation. To test the usefulness of the conjugate for biosensing, immunoglobulin G (IgG binding assay was conducted. The result showed that Ru(II-protein G conjugates were capable of binding IgG and the more cross-linkers to modify protein G, the higher conjugation efficiency. To demonstrate the feasibility of Ru(II-protein G conjugates for fluorescent immunoassays, the detection of recombinant histidine-tagged protein using the conjugates and anti-histidine antibody was developed. The results showed that the histidine-tagged protein was successfully detected with dose-response, indicating that Ru(II-protein G conjugate is a useful universal fluorescent reagent for quantitative immunoassays.

EIS immunosensor based on magnetic nanoparticles for ochratoxim A determination

International Nuclear Information System (INIS)

Biraruti, Irina; Tudorache, Madalina; Rotariu, Lucian; Bala, Camelia; Jaffrezic-Renault, Nicole

2009-01-01

Full text: Ochratoxin A (OTA) is a mycotoxin produced by Aspergillus and Penicillium fungi. It occurs in a variety of foodstuff, including cereals, coffee, wine, grape juice and fruits juice. Maximum permitted levels have been established by the EU and several countries. Currently, there are several methods available for OTA monitoring in real samples, e.g. TLC, HPLC, ELISA immunosensors. Liquid chromatography with fluorescence detection (HPLC-FLD), coupled with immunoaffinity column or solid phase extraction as pretreatment method, has been validated and adopted as official standard for OTA determination. Also, an immunosensor based on electrochemical detection has been reported for rapid analysis of OTA in food and beverages. Our work proposes the development of a new sensitive immunosensor combining the use of magnetic nanoparticles and EIS (electrochemical impedance spectroscopy) detection principle. Anti-OTA antibody immobilised on magnetic nanoparticles were deposited on the EIS electrode surface by applying a magnetic field. Then, OTA sample solutions were added in the detection cell. Quantitative determination of OTA content was performed based on EIS detection. Experimental parameters of the immunosensor were set-up at the optimum values. Based on those values, the immunosensor allows to detect OTA in the range 1-10 ng/mL. The future perspective is to validate this immunochemical method using HPLC-FLD. (authors)

Enzyme immunoassay of oestrogen receptors in needle biopsies from human liver

DEFF Research Database (Denmark)

Becker, U; Andersen, J; Poulsen, H S

1991-01-01

For quantitative assessments of sex hormone receptors in liver tissue, ligand binding assays are inconvenient, as they require large biopsies (0.5-1.0 g). The present study shows that it is possible to measure oestrogen receptors (ER) quantitatively in needle biopsy specimens as small as 10 mg...... by modifications of a commercial enzyme immunoassay employing monoclonal antibodies. Sucrose gradient centrifugation and the dextran charcoal method served as reference methods. A consecutive series of needle biopsies from patients suspected of liver disease were investigated. The biopsies (n = 37) had a median...

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.

Electrochemical photovoltaic cells and electrodes

Science.gov (United States)

Skotheim, Terje A.

1984-01-01

Improved electrochemical photovoltaic cells and electrodes for use therein, particularly electrodes employing amorphous silicon or polyacetylene coating are produced by a process which includes filling pinholes or porous openings in the coatings by electrochemical oxidation of selected monomers to deposit insulating polymer in the openings.

Electrochemical Hydrogen Evolution

DEFF Research Database (Denmark)

Laursen, A.B.; Varela Gasque, Ana Sofia; Dionigi, F.

2012-01-01

The electrochemical hydrogen evolution reaction (HER) is growing in significance as society begins to rely more on renewable energy sources such as wind and solar power. Thus, research on designing new, inexpensive, and abundant HER catalysts is important. Here, we describe how a simple experiment...... catalysts based on this. Suited for upper-level high school and first-year university students, this exercise involves using a basic two-cell electrochemical setup to test multiple electrode materials as catalysts at one applied potential, and then constructing a volcano curve with the resulting currents...

Electrochemical energy storage

CERN Document Server

Tarascon, Jean-Marie

2015-01-01

The electrochemical storage of energy has become essential in assisting the development of electrical transport and use of renewable energies. French researchers have played a key role in this domain but Asia is currently the market leader. Not wanting to see history repeat itself, France created the research network on electrochemical energy storage (RS2E) in 2011. This book discusses the launch of RS2E, its stakeholders, objectives, and integrated structure that assures a continuum between basic research, technological research and industries. Here, the authors will cover the technological

Electrochemical non-enzymatic glucose sensors

International Nuclear Information System (INIS)

Park, Sejin; Boo, Hankil; Chung, Taek Dong

2006-01-01

The electrochemical determination of glucose concentration without using enzyme is one of the dreams that many researchers have been trying to make come true. As new materials have been reported and more knowledge on detailed mechanism of glucose oxidation has been unveiled, the non-enzymatic glucose sensor keeps coming closer to practical applications. Recent reports strongly imply that this progress will be accelerated in 'nanoera'. This article reviews the history of unraveling the mechanism of direct electrochemical oxidation of glucose and making attempts to develop successful electrochemical glucose sensors. The electrochemical oxidation of glucose molecules involves complex processes of adsorption, electron transfer, and subsequent chemical rearrangement, which are combined with the surface reactions on the metal surfaces. The information about the direct oxidation of glucose on solid-state surfaces as well as new electrode materials will lead us to possible breakthroughs in designing the enzymeless glucose sensing devices that realize innovative and powerful detection. An example of those is to introduce nanoporous platinum as an electrode, on which glucose is oxidized electrochemically with remarkable sensitivity and selectivity. Better model of such glucose sensors is sought by summarizing and revisiting the previous reports on the electrochemistry of glucose itself and new electrode materials

Microfluidic electrochemical device and process for chemical imaging and electrochemical analysis at the electrode-liquid interface in-situ

Science.gov (United States)

Yu, Xiao-Ying; Liu, Bingwen; Yang, Li; Zhu, Zihua; Marshall, Matthew J.

2016-03-01

A microfluidic electrochemical device and process are detailed that provide chemical imaging and electrochemical analysis under vacuum at the surface of the electrode-sample or electrode-liquid interface in-situ. The electrochemical device allows investigation of various surface layers including diffuse layers at selected depths populated with, e.g., adsorbed molecules in which chemical transformation in electrolyte solutions occurs.

Repetitive Immunosensor with a Fiber-Optic Device and Antibody-Coated Magnetic Beads for Semi-Continuous Monitoring of Escherichia coli O157:H7.

Science.gov (United States)

Taniguchi, Midori; Saito, Hirokazu; Mitsubayashi, Kohji

2017-09-19

A rapid and reproducible fiber-optic immunosensor for Escherichia coli O157:H7 ( E. coli O157:H7) was described. The biosensor consisted of a flow cell, an optical fiber with a thin Ni layer, and a PC linked fluorometer. First, the samples with E. coli O157:H7 were incubated with magnetic beads coated with anti- E. coli O157:H7 antibodies and anti- E. coli O157:H7 antibodies labeled cyanine 5 (Cy5) to make sandwich complexes. Then the Cy5-( E. coli O157:H7)-beads were injected into a flow cell and pulled to the magnetized Ni layer on the optical fiber set in the flow cell. An excitation light (λ = 635 nm) was used to illuminate the optical fiber, and the Cy5 florescent molecules facing the optical fiber were exposed to an evanescent wave from the optical fiber. The 670 nm fluorescent light was measured using a photodiode. Finally, the magnetic intensity of the Ni layer was removed and the Cy5- E. coli O157:H7-beads were washed out for the next immunoassay. E. coli O157:H7, diluted with phosphate buffer (PB), was measured from 1 × 10⁵ to 1 × 10⁷ cells/mL. The total time required for an assay was less than 15 min (except for the pretreatment process) and repeating immunoassay on one optical fiber was made possible.

The Use of Recombinant Hemagglutinine Protein of Rinderpest Virus in Enzyme Immunoassay

OpenAIRE

BULUT, Hakan; BOLAT, Yusuf

2003-01-01

In this study, Rinderpest virus (RPV) recombinant hemagglutinine protein (rH) fused with protein A region of Staphylococcus aureus was expressed in Escherichia coli and purified by IgG affinity chromatography. rH protein was also used to establish enzyme immunoassay. Therefore, to prevent IgG binding to the protein A the wells coated with the rH proteins were blocked by human serum. Afterwards, RPV antigens were added to the wells to evaluate this assay. To this end, serum from mice immunized...

Thermodynamics of irreversible electrochemical phenomena

NARCIS (Netherlands)

Groot, S.R. de; Mazur, P.; Tolhoek, H.A.

1953-01-01

A discussion from first principles is given of the energy and entropy laws in electrochemical systems. It is found that it is possible to clarify such controversial concepts as the form of the second law and the role of the electrochemical potential in the systems concerned.

Accuracy of immunoassay and mass spectrometry urinary free cortisol in the diagnosis of Cushing's syndrome.

Science.gov (United States)

Aranda, G; Careaga, M; Hanzu, F A; Patrascioiu, I; Ríos, P; Mora, M; Morales-Romero, B; Jiménez, W; Halperin, I; Casals, G

2016-10-01

Urinary free cortisol (UFC) determination by highly specific methods as mass spectrometry instead of commercially available antibody-based immunoassays is increasingly recommended. However, clinical comparisons of both analytical approaches in the screening of Cushing's syndrome (CS) are not available. The aim of this study was to evaluate the diagnostic value of mass spectrometry versus immunoassay measurements of 24 h-UFC in the screening of CS. Cross-sectional study of 33 histologically confirmed CS patients: 25 Cushing's disease, 5 adrenal CS and 3 ectopic CS; 92 non-CS patients; and 35 healthy controls. UFC by immunoassay (UFCxIA) and mass spectrometry (UFCxMS), urinary free cortisone (UFCo) and UFC:UFCo ratio were measured, together with creatinine-corrected values. Sensitivity, specificity, AUC and Landis and Koch concordance index were determined. AUC for UFCxIA and UFCxMS were 0.77 (CI 0.68-0.87) and 0.77 (CI 0.67-0.87) respectively, with a kappa coefficient 0.60 and strong Landis and Koch concordance index. The best calculated cutoff values were 359 nmol/24 h for UFCxIA (78 % sensitivity, 62 % specificity) and 258.1 nmol/24 h for UCFxMS (53 % sensitivity, 86 % specificity). The upper limit of UFCxIA and UCFxMS reference ranges were 344.7 and 169.5 nmol/24 h respectively. Sensitivity and specificity for CS diagnosis at these cutpoints were 84 and 56 % for UFCxIA and 81 and 54 % for UFCxMS. According to our data, both methods present a very similar diagnostic value. However, results suggest that lower cutoff points for mass spectrometry may be necessary in order to improve clinical sensitivity.

Photoelectrochemical detection of enzymatically generated CdS nanoparticles: Application to development of immunoassay.

Science.gov (United States)

Barroso, Javier; Saa, Laura; Grinyte, Ruta; Pavlov, Valeri

2016-03-15

We report an innovative photoelectrochemical process (PEC) based on graphite electrode modified with electroactive polyvinylpyridine bearing osmium complex (Os-PVP). The system relies on the in situ enzymatic generation of CdS quantum dots (QDs). Alkaline phosphatase (ALP) catalyzes the hydrolisis of sodium thiophosphate (TP) to hydrogen sulfide (H2S) which in the presence Cd(2+) ions yields CdS semiconductor nanoparticles (SNPs). Irradiation of SNPs with the standard laboratory UV-illuminator (wavelength of 365 nm) results in photooxidation of 1-thioglycerol (TG) mediated by Os-PVP complex on the surface of graphite electrode at applied potential of 0.31 V vs. Ag/AgCl. A novel immunoassay based on specific enzyme linked immunosorbent assay (ELISA) combined with the PEC methodology was developed. Having selected the affinity interaction between bovine serum albumine (BSA) with anti-BSA antibody (AB) as a model system, we built the PEC immunoassay for AB. The new assay displays a linear range up to 20 ngmL(-1) and a detection limit (DL) of 2 ngmL(-1) (S/N=3) which is lower 5 times that of the traditional chromogenic ELISA test employing p-nitro-phenyl phosphate (pNPP). Copyright © 2015 Elsevier B.V. All rights reserved.

In Situ Investigation of Electrochemically Mediated Surface-Initiated Atom Transfer Radical Polymerization by Electrochemical Surface Plasmon Resonance.

Science.gov (United States)

Chen, Daqun; Hu, Weihua

2017-04-18

Electrochemically mediated atom transfer radical polymerization (eATRP) initiates/controls the controlled/living ATRP chain propagation process by electrochemically generating (regenerating) the activator (lower-oxidation-state metal complex) from deactivator (higher-oxidation-state metal complex). Despite successful demonstrations in both of the homogeneous polymerization and heterogeneous systems (namely, surface-initiated ATRP, SI-ATRP), the eATRP process itself has never been in situ investigated, and important information regarding this process remains unrevealed. In this work, we report the first investigation of the electrochemically mediated SI-ATRP (eSI-ATRP) by rationally combining the electrochemical technique with real-time surface plasmon resonance (SPR). In the experiment, the potential of a SPR gold chip modified by the self-assembled monolayer of the ATRP initiator was controlled to electrochemically reduce the deactivator to activator to initiate the SI-ATRP, and the whole process was simultaneously monitored by SPR with a high time resolution of 0.1 s. It is found that it is feasible to electrochemically trigger/control the SI-ATRP and the polymerization rate is correlated to the potential applied to the gold chip. This work reveals important kinetic information for eSI-ATRP and offers a powerful platform for in situ investigation of such complicated processes.

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. Copyright © 2015 Elsevier B.V. All rights reserved.

Plasma exchange to remove HIT antibodies: dissociation between enzyme-immunoassay and platelet activation test reactivities.

Science.gov (United States)

Warkentin, Theodore E; Sheppard, Jo-Ann I; Chu, F Victor; Kapoor, Anil; Crowther, Mark A; Gangji, Azim

2015-01-01

Repeated therapeutic plasma exchange (TPE) has been advocated to remove heparin-induced thrombocytopenia (HIT) IgG antibodies before cardiac/vascular surgery in patients who have serologically-confirmed acute or subacute HIT; for this situation, a negative platelet activation assay (eg, platelet serotonin-release assay [SRA]) has been recommended as the target serological end point to permit safe surgery. We compared reactivities in the SRA and an anti-PF4/heparin IgG-specific enzyme immunoassay (EIA), testing serial serum samples in a patient with recent (subacute) HIT who underwent serial TPE precardiac surgery, as well as for 15 other serially-diluted HIT sera. We observed that post-TPE/diluted HIT sera-when first testing SRA-negative-continue to test strongly positive by EIA-IgG. This dissociation between the platelet activation assay and a PF4-dependent immunoassay for HIT antibodies indicates that patients with subacute HIT undergoing repeated TPE before heparin reexposure should be tested by serial platelet activation assays even when their EIAs remain strongly positive. © 2015 by The American Society of Hematology.

Development of an Ultrasensitive Immunoassay for Detecting Tartrazine

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Chuanlai Xu

2013-06-01

Full Text Available We have developed an ultrasensitive indirect competitive enzyme-linked immunosorbent assay for the determination of tartrazine. Two carboxylated analogues of tartrazine with different spacer lengths, and one derivative from commercial tartrazine after a little chemical modification, were synthesized as haptens in order to produce antibodies specific to tartrazine. The effect of sulfonic acid groups on the hapten structure of tartrazine was also studied carefully for the first time. A most specific monoclonal antibody against tartrazine was created and exhibited an IC50 value of 0.105 ng/mL and a limit of detection of 0.014 ng/mL, with no cross-reactivity to other structurally-related pigments. The established immunoassay was applied to the determination of tartrazine in fortified samples of orange juice and in real positive samples of carbonated beverages.

New antibody and immunoassay pretreatment strategy to screen polychlorinated biphenyls in Korean transformer oil.

Science.gov (United States)

Terakado, Shingo; Ohmura, Naoya; Park, Seok-Un; Lee, Seung-Min; Glass, Thomas R

2013-01-01

Development and modifications are described that expand the application of an immunoassay from the detection of Kanechlors (Japanese technical PCBs mixtures) to the detection of Aroclors (U. S. technical PCB mixtures, used in Korea) in contaminated Korean transformer oil. The first necessary modification was the development of a new antibody with a reactivity profile favorable for Aroclors. The second modification was the addition of a second column to the solid-phase extraction method to reduce assay interference caused by the Korean oil matrix. The matrix interference is suspected to be caused by the presence of synthetic oils (or similar materials) present as contaminants. The modified assay was validated by comparison to high-resolution gas chromatography/high-resolution mass spectrometry analysis, and was shown to be tolerant of up to 10% of several common synthetic insulating oils. Finally the screening performance of the modified assay was evaluated using 500 used transformer oil samples of Korean origin, and was shown to have good performance in terms of false positive and false negative rates. This report provides evidence for the first establishment of immunoassay screening for Aroclor based PCB contamination in Korean transformer oil.

Electrochemical impedance spectroscopy of oxidized porous silicon

Energy Technology Data Exchange (ETDEWEB)

Mula, Guido, E-mail: guido.mula@unica.it [Dipartimento di Fisica, Università degli Studi di Cagliari, Cittadella Universitaria di Monserrato, S.P. 8 km 0.700, 09042 Cagliari (Italy); Tiddia, Maria V. [Dipartimento di Fisica, Università degli Studi di Cagliari, Cittadella Universitaria di Monserrato, S.P. 8 km 0.700, 09042 Cagliari (Italy); Ruffilli, Roberta [Nanochemistry, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genova (Italy); Falqui, Andrea [Nanochemistry, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genova (Italy); Dipartimento di Scienze Chimiche e Geologiche, Università degli Studi di Cagliari, Cittadella Universitaria di Monserrato, S.P. 8 km 0.700, 09042 Cagliari (Italy); Palmas, Simonetta; Mascia, Michele [Dipartimento di Ingegneria Meccanica Chimica e dei Materiali, Università degli Studi di Cagliari, Piazza d' Armi, 09126 Cagliari (Italy)

2014-04-01

We present a study of the electrochemical oxidation process of porous silicon. We analyze the effect of the layer thickness (1.25–22 μm) and of the applied current density (1.1–11.1 mA/cm{sup 2}, values calculated with reference to the external samples surface) on the oxidation process by comparing the galvanostatic electrochemical impedance spectroscopy (EIS) measurements and the optical specular reflectivity of the samples. The results of EIS were interpreted using an equivalent circuit to separate the contribution of different sample parts. A different behavior of the electrochemical oxidation process has been found for thin and thick samples: whereas for thin samples the oxidation process is univocally related to current density and thickness, for thicker samples this is no more true. Measurements by Energy Dispersive Spectroscopy using a Scanning Electron Microscopy confirmed that the inhomogeneity of the electrochemical oxidation process is increased by higher thicknesses and higher currents. A possible explanation is proposed to justify the different behavior of thin and thick samples during the electrochemical process. - Highlights: • A multidisciplinary approach on porous Si electrochemical oxidation is proposed. • Electrochemical, optical, and structural characterizations are used. • Layer thickness and oxidation current effects are shown. • An explanation of the observed behavior is proposed.

Electrochemical polymer electrolyte membranes

CERN Document Server

Fang, Jianhua; Wilkinson, David P

2015-01-01

Electrochemical Polymer Electrolyte Membranes covers PEMs from fundamentals to applications, describing their structure, properties, characterization, synthesis, and use in electrochemical energy storage and solar energy conversion technologies. Featuring chapters authored by leading experts from academia and industry, this authoritative text: Discusses cutting-edge methodologies in PEM material selection and fabricationPoints out important challenges in developing PEMs and recommends mitigation strategies to improve PEM performanceAnalyzes the cur

A redox-mediated chromogenic reaction and application in immunoassay.

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Yu, Ru-Jia; Ma, Wei; Peng, Mao-Pan; Bai, Zhi-Shan; Long, Yi-Tao

2016-08-31

A novel redox-mediated chromogenic reaction was demonstrated based on the reaction between HAuCl4 and 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), which generate various color responses from red to green in the resulting solutions. Various redox substance could be used to mediate the reaction and trigger a distinct color response. We established a sensitive hydrogen peroxide colorimetric sensor based on the redox-mediated chromogenic reaction and depicted the application both in detection of enzyme and in an immunoassay. Combining the traditional chromogenic reagent with gold nanoparticles, our assay has the advantage in short response time (within three minutes), high sensitivity (10(-12) g mL(-1) for HBsAg) and stability. Copyright © 2016. Published by Elsevier B.V.

Performance of immunoassay kits for site characterization and remediation

International Nuclear Information System (INIS)

Waters, L.C.; Palausky, A.; Counts, R.W.; Jenkins, R.A.

1995-01-01

The US Department of Energy (DOE) is supporting efforts to identify, validate and implement the use of effective, low-cost alternatives to currently used analytical methods for environmental management. As part of that program, we have evaluated the performances of a number of immunoassay (IA) kits with specificities for environmental contaminants of concern to the DOE. The studies were done in the laboratory using both spiked and field test samples. The analyte specificity and manufacturers of the kits evaluated were the following: mercury, BioNebraska; polychlorinated biphenyls (PCBs), EnSys and Millipore; petroleum fuel hydrocarbons, Millipore and Ohmicron; and polyaromatic hydrocarbons (PAHs), Ohmicron and Millipore. The kits were used in either a semiquantitative or quantitative format according to the preference of the manufacturers

A novel fluorescence immunoassay for the sensitive detection of Escherichia coli O157:H7 in milk based on catalase-mediated fluorescence quenching of CdTe quantum dots.

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Chen, Rui; Huang, Xiaolin; Li, Juan; Shan, Shan; Lai, Weihua; Xiong, Yonghua

2016-12-01

Immunoassay is a powerful tool for rapid detection of food borne pathogens in food safety monitoring. However, conventional immunoassay always suffers from low sensitivity when it employs enzyme-catalyzing chromogenic substrates to generate colored molecules as signal outputs. In the present study, we report a novel fluorescence immunoassay for the sensitive detection of E. coli O157:H7 through combination of the ultrahigh bioactivity of catalase to hydrogen peroxide (H 2 O 2 ) and H 2 O 2 -sensitive mercaptopropionic acid modified CdTe QDs (MPA-QDs) as a signal transduction. Various parameters, including the concentrations of anti-E. coli O157:H7 polyclonal antibody and biotinylated monoclonal antibody, the amounts of H 2 O 2 and streptavidin labeled catalase (CAT), the hydrolysis temperature and time of CAT to H 2 O 2 , as well as the incubation time between H 2 O 2 and MPA-QDs, were systematically investigated and optimized. With optimal conditions, the catalase-mediated fluorescence quenching immunoassay exhibits an excellent sensitivity for E. coli O157:H7 with a detection limit of 5 × 10 2  CFU/mL, which was approximately 140 times lower than that of horseradish peroxidase-based colorimetric immunoassay. The reliability of the proposed method was further evaluated using E. coli O157:H7 spiked milk samples. The average recoveries of E. coli O157:H7 concentrations from 1.18 × 10 3  CFU/mL to 1.18 × 10 6  CFU/mL were in the range of 65.88%-105.6%. In brief, the proposed immunoassay offers a great potential for rapid and sensitive detection of other pathogens in food quality control. Copyright © 2016 Elsevier B.V. All rights reserved.

Strip-based immunoassay for the simultaneous detection of the neonicotinoid insecticides imidacloprid and thiamethoxam in agricultural products

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A semiquantitative strip immunoassay was developed for the rapid detection of imidacloprid and thiamethoxam in agricultural products using specific nanocolloidal gold-labeled monoclonal antibodies. The conjugates of imidacloprid-BSA and thiamethoxam-BSA and goat anti-mouse IgG were coated on the ni...

Quantification of patient specific assay interference in different formats of enzyme linked immunoassays for therapeutic monoclonal antibodies

NARCIS (Netherlands)

Grebenchtchikov, N.J.; Geurts-Moespot, A.; Heijmen, L.; Laarhoven, H.W.M. van; Herpen, C.M.L. van; Thijs, A.M.J.; Span, P.N.; Sweep, F.C.

2014-01-01

BackgroundThe use of therapeutic monoclonal antibodies for clinical purposes has significantly increased in recent years, and so has the need to monitor antibody concentrations. This may be achieved using the well-established enzyme linked immunoassay (ELISA) methods; however, these assays are

Correlation between morphology and magnetic properties of electrochemically produced cobalt powder particles

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Maksimović Vesna M.

2015-01-01

Full Text Available Cobalt 3D powder particles were successfully prepared by the galvanostatic electrodeposition. Electrodeposited cobalt powder were characterized by X-ray diffraction (XRD, scanning electron microscope (SEM, Energy Dispersive Spectroscopy (EDS analysis and SQUID magnetometry. It has been shown that morphology, structure and magnetic properties of cobalt particles are closely associated and can be easily controlled by adjusting process parameters of electrodeposition. Morphology of cobalt powder particles is strongly affected by hydrogen evolution reaction as a parallel reaction to cobalt electrodeposition. Depending on the applied current density, the two types of powder particles were formed: dendrites at lower and spongy-like particles at higher current densities. Morphologies and structures of powder particles are correlated with their magnetic properties, and compared with those of the bulk cobalt. In comparison with the properties of bulk cobalt, the obtained 3D structures exhibited a decreased saturation magnetization (MS, but an enhanced coercivity (HC which is explained by their peculiar morphology. [Projekat Ministarstva nauke Republike Srbije, br. III 45012

Analytical validation of Gentian NGAL particle-enhanced enhanced turbidimetric immunoassay (PETIA

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Gian Luca Salvagno

2017-08-01

Full Text Available Objectives: This study was designed to validate the analytical performance of the new Gentian particle-enhanced enhanced turbidimetric immunoassay (PETIA for measuring neutrophil gelatinase-associated lipocalin (NGAL in serum samples. Design and methods: Analytical validation of the Gentian NGAL assay was carried out on a Roche Cobas c501 and was based on assessment of limit of blank (LOB, limit of detection (LOD, functional sensitivity, imprecision, linearity and concordance with the BioPorto NGAL test. Results: The LOB and LOD of Gentian NGAL were found to be 3.8 ng/mL and 6.3 ng/mL, respectively. An analytical coefficient of variation (CV of 20% corresponded to a NGAL value of 10 ng/mL. The intra-assay and inter-assay imprecision (CV was between 0.4 and 5.2% and 0.6 and 7.1% and the total imprecision (CV was 3.7%. The linearity was optimal at NGAL concentrations between 37 and 1420 ng/mL (r=1.00; p<0.001. An excellent correlation was observed between values measured with Gentian NGAL and BioPorto NGAL in 74 routine serum samples (r=0.993. The mean percentage bias of the Gentian assay versus the Bioporto assay was +3.1% (95% CI, +1.6% to +4.5%. Conclusions: These results show that Gentian NGAL may be a viable option to other commercial immunoassays for both routine and urgent assessment of serum NGAL. Keywords: Neutrophil gelatinase-associated lipocalin, NGAL, Analytical validation, Acute kidney injury

A novel fluorescence immunoassay for the sensitive detection of Escherichia coli O157:H7 in milk based on catalase-mediated fluorescence quenching of CdTe quantum dots

International Nuclear Information System (INIS)

Chen, Rui; Huang, Xiaolin; Li, Juan; Shan, Shan; Lai, Weihua; Xiong, Yonghua

2016-01-01

Immunoassay is a powerful tool for rapid detection of food borne pathogens in food safety monitoring. However, conventional immunoassay always suffers from low sensitivity when it employs enzyme-catalyzing chromogenic substrates to generate colored molecules as signal outputs. In the present study, we report a novel fluorescence immunoassay for the sensitive detection of E. coli O157:H7 through combination of the ultrahigh bioactivity of catalase to hydrogen peroxide (H_2O_2) and H_2O_2-sensitive mercaptopropionic acid modified CdTe QDs (MPA-QDs) as a signal transduction. Various parameters, including the concentrations of anti-E. coli O157:H7 polyclonal antibody and biotinylated monoclonal antibody, the amounts of H_2O_2 and streptavidin labeled catalase (CAT), the hydrolysis temperature and time of CAT to H_2O_2, as well as the incubation time between H_2O_2 and MPA-QDs, were systematically investigated and optimized. With optimal conditions, the catalase-mediated fluorescence quenching immunoassay exhibits an excellent sensitivity for E. coli O157:H7 with a detection limit of 5 × 10"2 CFU/mL, which was approximately 140 times lower than that of horseradish peroxidase-based colorimetric immunoassay. The reliability of the proposed method was further evaluated using E. coli O157:H7 spiked milk samples. The average recoveries of E. coli O157:H7 concentrations from 1.18 × 10"3 CFU/mL to 1.18 × 10"6 CFU/mL were in the range of 65.88%–105.6%. In brief, the proposed immunoassay offers a great potential for rapid and sensitive detection of other pathogens in food quality control. - Highlights: • A novel fluorescence immunoassay was developed for the ultrasensitive detection of E. coli O157:H7. • This detection was achieved through the combination of the high bioactivity of CAT and H_2O_2-sensitive QDs. • The activity of CAT to H_2O_2 is 1000 folds higher than that of the HRP to tetramethylbenzidine. • The limit of detection of the proposed method could

Pediatric reference value distributions and covariate-stratified reference intervals for 29 endocrine and special chemistry biomarkers on the Beckman Coulter Immunoassay Systems: a CALIPER study of healthy community children.

Science.gov (United States)

Karbasy, Kimiya; Lin, Danny C C; Stoianov, Alexandra; Chan, Man Khun; Bevilacqua, Victoria; Chen, Yunqi; Adeli, Khosrow

2016-04-01

The CALIPER program is a national research initiative aimed at closing the gaps in pediatric reference intervals. CALIPER previously reported reference intervals for endocrine and special chemistry markers on Abbott immunoassays. We now report new pediatric reference intervals for immunoassays on the Beckman Coulter Immunoassay Systems and assess platform-specific differences in reference values. A total of 711 healthy children and adolescents from birth to reference intervals calculated in accordance with Clinical and Laboratory Standards Institute (CLSI) EP28-A3c guidelines. Complex profiles were observed for all 29 analytes, necessitating unique age and/or sex-specific partitions. Overall, changes in analyte concentrations observed over the course of development were similar to trends previously reported, and are consistent with biochemical and physiological changes that occur during childhood. Marked differences were observed for some assays including progesterone, luteinizing hormone and follicle-stimulating hormone where reference intervals were higher than those reported on Abbott immunoassays and parathyroid hormone where intervals were lower. This study highlights the importance of determining reference intervals specific for each analytical platform. The CALIPER Pediatric Reference Interval database will enable accurate diagnosis and laboratory assessment of children monitored by Beckman Coulter Immunoassay Systems in health care institutions worldwide. These reference intervals must however be validated by individual labs for the local pediatric population as recommended by CLSI.

Electrochemical properties of ion implanted silicon

International Nuclear Information System (INIS)

Pham minh Tan.

1979-11-01

The electrochemical behaviour of ion implanted silicon in contact with hydrofluoric acid solution was investigated. It was shown that the implanted layer on silicon changes profoundly its electrochemical properties (photopotential, interface impedance, rest potential, corrosion, current-potential behaviour, anodic dissolution of silicon, redox reaction). These changes depend strongly on the implantation parameters such as ion dose, ion energy, thermal treatment and ion mass and are weakly dependent on the chemical nature of the implantation ion. The experimental results were evaluated and interpreted in terms of the semiconductor electrochemical concepts taking into account the interaction of energetic ions with the solid surface. The observed effects are thus attributed to the implantation induced damage of silicon lattice and can be used for profiling of the implanted layer and the electrochemical treatment of the silicon surface. (author)

A direct radio-immunoassay for plasma aldosterone: significance of endogenous cortisol

International Nuclear Information System (INIS)

Man, A.J.M. de; Hofman, J.A.; Hendriks, Th.; Rosmalen, F.M.A.; Ross, H.A.; Benraad, Th.J.

1980-01-01

A direct radio-immunoassay for plasma aldosterone was developed, using a highly specific antiserum raised in sheep. An excellent correlation was observed between its results and the levels measured after extraction and chromatography. The necessity of including a blocking agent to inhibit the binding of aldosterone to plasma proteins was investigated. It was found that in low-cortisol ( 10 μg/100 ml) the final assay result was independent of the presence of ANS. The effect of cortisol was interpreted in terms of its influence on aldosterone binding to plasma proteins in the absence of a blocking agent. (Auth.)

Quantification of patient-specific assay interference in different formats of enzyme-linked immunoassays for therapeutic monoclonal antibodies

NARCIS (Netherlands)

Grebenchtchikov, Nicolai; Geurts-Moespot, Anneke J.; Heijmen, Linda; van Laarhoven, Hanneke W. M.; van Herpen, Carla M. L.; Thijs, Annemarie M. J.; Span, Paul N.; Sweep, Fred C. G. J.

2014-01-01

The use of therapeutic monoclonal antibodies for clinical purposes has significantly increased in recent years, and so has the need to monitor antibody concentrations. This may be achieved using the well-established enzyme linked immunoassay (ELISA) methods; however, these assays are subject to a

DEMONSTRATION BULLETIN: PCP IMMUNOASSAY TECHNOLOGIES - PENTA RISC BY ENSYS INC., PENTA RAPID BY OHMICRON CORP., ENVIROGARD BY MILLIPORE

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