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Functionally Fused Antibodies - a novel adjuvant fusion system
2008-01-01
Immunohistochemical diagnosis of systemic bovine zygomycosis by murine monoclonal antibodies
1996-01-01
Development of murine monoclonal antibodies for the immunohistochemical diagnosis of systemic bovine aspergillosis
1996-01-01
Development of murine monoclonal antibodies for the immunohistochemical diagnosis of systemic bovine aspergillosis
1996-01-01
Role of Antibodies in Cognitive Dysfunction in Patients With Systemic Lupus Erythematosus
Systemic Lupus Erythematosus
1989-01-01
Two major areas of application of monoclonal antibodies were examined: the development of products to support the 'Antibody Delivery System', a parent-specific and variable antibody formula drug system for use in imaging and treatment of cancer, and the development of an antibody-based radiopharmaceutical for imaging occult abscesses and other conditions involving high concentrations of white blood cells. In development of the Antibody Delivery System components, methods for characterization and purification of monoclonal antibodies were developed and validated. A dot immunoassay test, under the name RhoDot (TM) Immunoassay, was developed for matching antibodies to putative tumor specimen: a radioimmunoassay, under the name PhoChek (TM) Quality Control Test Kit for Radiolabeled Antibodies, was developed and commercialized for measuring the immunoreactive ... >>
Radiolabeled Monoclonal Antibody Therapy in Treating Patients With Primary Brain Tumors
Brain and Central Nervous System Tumors
Comparison of intramuscular therapy with Erwinia asparaginase and asparaginase Medac: pharmacokinetics, pharmacodynamics, formation of antibodies and influence on the coagulation system
2001-01-01
Comparison of intramuscular therapy with Erwinia asparaginase and asparaginase Medac : Pharmacokinetics, pharmacodynamics, formation of antibodies and influence on the coagulation system
2001-01-01
Comparison of intramuscluar therapy with Erwinia asparaginase and asparaginase and asparaginase Medac: pharmacokinetics, pharmacodynamics, formation of antibodies and influence on the coagulation system
2002-01-01
Fluorogenic Cell-Based Biosensors for Monitoring Microbes
... to work when utilizing either immunoglobulin E (IgE) antibodies or traditionally generated rat antibodies - a promising result in that it indicates that the systems could be ...
Brain and Central Nervous System Tumors; Lymphoma
1989-04-01
Two major areas of application of monoclonal antibodies were examined: the development of products to support the 'Antibody Delivery System', a parent-specific and variable antibody formula drug system for use in imaging and treatment of cancer, and the development of an antibody-based radiopharmaceutical for imaging occult abscesses and other conditions involving high concentrations of white blood cells. In development of the Antibody Delivery System components, methods for characterization and purification of monoclonal antibodies were developed and validated; a dot immunoassay test, under the name RhoDot (TM) Immunoassay, was developed for matching antibodies to putative tumor specimen: a radioimmunoassay, under the name PhoChek (TM) Quality Control Test Kit for Radiolabeled Antibodies, was developed and commercialized for measuring the immunoreactive fraction of radiolabeled antibodies specific to colorecal cancer; and a patient-specific quality control test was developed. In development of the antibody-based radiopharmaceutical for imaging occult abscesses, a candidate antibody was identified and produced under U.S. Food and Drug Administration standards preparatory to human clinical trials.
1987-01-01
The usefulness of radiolabeled monoclonal antibodies for imaging and treatment of human (ovarian) cancer was investigated. A review of tumor imaging with monoclonal antibodies is presented. Special attention is given to factors that influence the localization of the antibodies in tumors, isotope choice and methods of radiolabeling of the monoclonal antibodies. Two monoclonal antibodies, OC125 and OV-TL3, with high specificity for human epithelial ovarian cancer are characterized. A simple radio-iodination technique was developed for clinical application of the monoclonal antibodies. The behavior of monoclonal antibodies in human tumor xenograft systems and in man are described. Imaging of tumors is complicated because of high background levels of radioactivity in other sites than the tumor, especially in the bloodpool. A technique was developed to improve ... >>
1984-01-01
A solid-phase immunosorbent radioassay for the detection of circulating antibodies to protein hormones is described. The assay is based on the binding of the homologous 125I-labelled antigen to the antibodies which are then bound to anti-IgG antibodies covalently coupled to Sepharose. It can easily be applied as a complement to any radioimmunoassay for the detection of circulating antibodies to the ligand measured. The assay system avoids falsely elevated values due to interference of high serum concentrations of the antigen. The assay was applied to measure antibodies to FSH, LH, TSH, GH, prolactin, insulin and thyroglobulin (Tg). Among patients with chronic thyroiditis Tg antibodies were found in 100% of the sera. In diffuse toxic goitre 73% of the patients had detectable Tg antibodies. Insulin antibodies were present in 82% of the sera from patients with ... >>
Affinity of antibody secreted by a single cell. [Radioimmunoassay of human serum albumin]
1978-01-01
It was the intention of this research to measure the affinity of antibody secreted by a single cell, and to describe the spectrum of affinities displayed in response to antigenic stimulation. The single cell secreting specific antibody was isolated by means of the hemolytic plaque assay. The amount of antibody secreted by the cell was to be measured through the use of a solid phase radioimmunoassay. The affinity of the antibody would be estimated by comparing the diameter of the plaque, and the amount of antibody secreted, with a mathematical theory of the formation of a plaque in agar. As a test system, a solid phase radioimmunoassay was developed for human serum albumin using antibody coupled to Sephadex. A sensitivity of 1 nanogram was attained with this assay. A solid phase radioimmunoassay for mouse immunoglobulin M was developed, using antibody coupled to Sepharose. The sensitivity attained with this assay was only on the order of 10 micrograms. The mouse immunoglobulin M radioimmunoassay was not sensitive enough to measure the amount of antibody secreted by a single cell. From a theoretical equation, the relationship between antibody affinity, plaque diameter and antibody secretion rate was calculated for the experimental conditions used in this research. By assuming a constant antibody secretion rate, an effective binding constant for the antibody was estimated from the average plaque diameters. This effective binding constant was observed to increase during the immune response.
Monoclonal antibodies as physiologic probes
1983-01-01
This report covers four independent studies exploring biomedical applications of monoclonal antibodies. The anti-digoxin hybridoma system illustrates: (1) a series of different monoclonal antibodies to a simple hapten can serve as a model system for studying the fine specificity of the antigen-antibody combining site. (2) high-affinity monoclonal antibody can serve as a reliable reagent in a clinical assay for the determination of drug levels such as digoxin in patients' sera. And (3) Fab prepared from monoclonal anti-digoxin antibody may serve as a therapeutic drug for the reversal of digoxin intoxication. Monoclonal anti-mullerian inhibiting substance (anti-MIS) antibody will facilitate the purification of MIS by affinity chromatography. Monoclonal anti-MIS antibodies will aid in the detection of both the site of cellular production of MIS and the receptors for ... >>
1976-12-01
Antibodies to ultraviolet light denatured DNA (UV DNA) have been measured in patients with systemic lupus erythematosus (SLE) and normal subjects, using a millipore filter radioimmunoassay. High levels of UV DNA binding were only found in patients with SLE. The presence of UV DNA antibodies correlated well with the presence of native DNA antibodies, although immunodiffusion studies and inhibition techniques showed these antibodies to be immunologically distinct in many cases. Forty-one percent of the SLE patients had had photosensitivity at some stage of their disease, but there was a poor correlation between this symptom and the presence of UV DNA antibodies. Although UV DNA is known to be a potent immunogen, none of the results from this study suggests that antibodies to UV DNA are more than another example of the broad spectrum of antinuclear antibodies seen in SLE.
Anti-chromatin and anti-histone antibodies in Egyptian patients with systemic lupus erythematosus
2009-01-01
There has been a renewed interest in anti-chromatin and anti-histone antibodies in the last few years. To assess the prevalence of anti-chromatin and anti-histone antibodies in patients with systematic lupus erythematosus (SLE) and to correlate serum levels of these antibodies with clinical features of the disease, the presence of anti-chromatin and anti-histone antibodies in 38 patients with SLE was investigated by an enzyme-linked immunosorbent assay (ELISA). To determine the specificity of these antibodies, 15 patients with rheumatoid arthritis, 15 patients with systemic sclerosis, and 15 normal controls were also tested. Sensitivity of anti-chromatin antibodies in SLE patients was 89.5% and specificity was 80.0%, while sensitivity of anti-histone antibodies was 92.1% and specificity wa...
Sensitive radioimmunoassay for the detection of monoclonal anti-idiotype antibodies
A radioimmunoassay was developed in order to detect anti-idiotypic antibodies in the supernatants of hybrid cells. This assay is both sensitive and specific for anti-idiotypic (but not anti-allotypic) antibodies. Monoclonal antibodies present in test supernatants are bound by an anti-immunoglobulin coated solid phase. Subsequent incubation with a source of mouse immunoglobulin 'blocks' unreacted anti-immunoglobulin antibodies on the solid phase. Anti-idiotypic antibodies are then detected by their ability to bind /sup 125/I-labelled idiotype-bearing antibody. This paper describes the use of this assay to detect monoclonal anti-idiotypic antibodies in 2 systems; the cross-reactive idiotype of A/J anti-ABA antibodies, and the idiotype expressed by the myeloma protein HOPC 8. Similarly, /sup 125/I-labelled anti-idiotype antibodies may be used in this assay to detect monoclonal idiotype-bearing antibodies. Further modifications are described which would allow the detection of monoclonal anti-allotype antibodies.
Sensitive radioimmunoassay for the detection of monoclonal anti-idiotype antibodies
1983-02-25
A radioimmunoassay was developed in order to detect anti-idiotypic antibodies in the supernatants of hybrid cells. This assay is both sensitive and specific for anti-idiotypic (but not anti-allotypic) antibodies. Monoclonal antibodies present in test supernatants are bound by an anti-immunoglobulin coated solid phase. Subsequent incubation with a source of mouse immunoglobulin 'blocks' unreacted anti-immunoglobulin antibodies on the solid phase. Anti-idiotypic antibodies are then detected by their ability to bind /sup 125/I-labelled idiotype-bearing antibody. This paper describes the use of this assay to detect monoclonal anti-idiotypic antibodies in 2 systems; the cross-reactive idiotype of A/J anti-ABA antibodies, and the idiotype expressed by the myeloma protein HOPC 8. Similarly, /sup 125/I-labelled anti-idiotype antibodies may be used in this assay to detect monoclonal idiotype-bearing antibodies. Further modifications are described which would allow the detection of monoclonal anti-allotype antibodies.
2009-01-01
The ability of anti-D antibodies to cause antigen-specific immunosuppression depends on their interaction with low-affinity Fc?-receptors. Human monoclonal antibodies to D antigen of the rhesus system were investigated by antibody-dependent cytotoxicity assay in order to estimate their ability to induce hemolysis mediated by low-affinity Fc? receptors. We demonstrate that affinity of monoclonal antibodies to receptors of this type does not depend on primary structure of Fc-fragment, but depends on the producer cell line which expresses the antibodies. Monoclonal IgG1 antibodies interacting with Fc?RIIa and Fc?RIII lost this property, if they were secreted by human-mouse heterohybridoma, but not by human B-cell line. On the opposite, monoclonal antibodies that could not activate low-affinit...
1984-03-01
Full Text Available.The binding properties of an immune complex-forming system comprising human IgG and mouse monoclonal antibodies against human IgG have been studied. A refined binding assay has been applied directly on ascitic fluid containing monoclonal antibody. Complete sets of binding data of a series of different monoclonal antibodies were collected and analysed by various graphical and statistical methods. Special attention was given to methods which allow determination of specific monoclonal antibody concentration as well as antibody affinity. It was found that the formation of genuine antigen: antibody complexes per se gives rise to deviations from expected linearity in commonly used binding equations. Good correlation was found between the antibody concentrations obtained by various graphical approaches, whereas the size of the association constant seemed to depend on the method in use. The binding pattern was found to be dependent on the concentration of antibody. Most reliable parameters were obtained if the product of the antibody concentration and the association constant was below 10.
Exploiting the diversity of time scales in the immune system: A B-cell antibody model
1991-06-01
Using the continuous shape-space formalism, the authors develop an immune system model involving both B lymphocytes and antibody molecules. The binding and cross-linking of receptors on B cells stimulates the cells to divide and, with a lag, to secrete antibody. Using the method of multiple scales, it is shown how to correctly formulate long-time-scale equations for the population dynamics of B cells, the total antibody concentration, and rate of antibody secretion. The authors model is compared with previous phenomenological formulations.
Kinetic Studies of a Doubly Bound Red Cell Antigen-Antibody System
1972-08-01
Full Text Available. The Polybrene method for detection of red cell antibodies which utilizes continuous flow equipment was modified so that kinetic studies could be performed on red cell antibodies doubly bound between adjacent red cells. In the anti-Rho-Rho erythrocyte system, deaggregation by temperature was studied over an antibody concentration range of from approximately 1 to 500 antibody molecules per erythrocyte, a residence time range of approximately eightfold, and a temperature range of from 10 to 55°C. The rate of dissociation of antigen-antibody complex, as determined from deaggregation of antibody-dependent red cell aggregates, was found to be of apparent zero order. The apparent activation energy for the antigen-antibody reaction under the experimental conditions was determined and found to be higher than would be expected for singly bound antigen-antibody systems. Possible explanations are considered for these findings in terms of an antigen-antibody bond-breaking model.
Monoclonal antibodies in clinical and experimental oncology. Imaging and idiotypic vaccines
1987-01-01
This paper reports on two topics. Both are centered on monoclonal antibody-defined human tumor-associated antigens. The first is a totally applicative effort to evaluate the clinical usefulness of the radiolabelled monoclonal antibodies as in vivo diagnostic tools and the second is a laboratory research project to manipulate the immune system of mice and rabbits to produce anti-tumor responses using monoclonal antibodies as idiotypic vaccines
Heterophyle antibodies causing false positive radio-immunoassay results a case report
1987-01-01
A case report of falsely elevated serum hormone values measured by radio-immunoassay (RIA) is described. The radio-immunoassays concerned have a first antibody raised in rabbits and mostly a separation technique based on a second antibody-solid phase system. The presence of heterophyle (anti-rabbit) antibodies in patients' serum is proved.
Heterophyle antibodies causing false positive radio-immunoassay results a case report
1987-01-01
A case report of falsely elevated serum hormone values measured by radio-immunoassay (RIA) is described. The radio-immunoassays concerned have a first antibody raised in rabbits and mostly a separation technique based on a second antibody-solid phase system. The presence of heterophyle (anti-rabbit) antibodies in patients' serum is proved
Fv antibodies to aflatoxin B1 derived from a pre-immunized antibody phage display library system
The production and characterization of recombinant antibodies to aflatoxin B[SUB1] (AFB[SUB1]), a potent mycotoxin and carcinogen is described. The antibody fragments produced were then applied for us ... Full Text Available
Digital Repository Infrastructure Vision for European Research (DRIVER)
1988-01-01
This book contains 8 chapters. Some of the titles are: Anti-DNA Antibodies in SLE: Historical Perspective; Specificity of Anti-DNA Antibodies in Systemic Lupus Erythematosus; Monoclonial Autoimmune Anti-DNA Antibodies; and Structure--Function Analyses of Anti-DNA Autoantibodies.
1984-02-01
Sodium ethylmercurithiosalicylate (Merthiolate), used for preservation of buffer solutions, was found to inhibit the antigen-antibody binding reactions in radioimmunoassay systems in a noncompetitive manner. This effect is attributable to mercury ion per se. 10 references. 2 figures.
Recall of ORTHO Antibody to HBsAg ELISA Test System 3-Ortho ...
... failures when using ORTHO Antibody to HBsAg ELISA Test System 3. This is due to low Optical Density (OD ... Division of Communication and Consumer Affairs. ...
Antibody Monitoring System Class 1 and Class II (AMS1 and AMS1+2)
... Cosmetics; Radiation-Emitting Products; Tobacco Products. -. Vaccines, Blood & Biologics. ... Antibody Monitoring System Class 1 and Class II (AMS1 and AMS1+2). ...
Brain and Central Nervous System Tumors; Metastatic Cancer; Neuroblastoma
Brain and Central Nervous System Tumors; Metastatic Cancer
Brain and Central Nervous System Tumors; Neuroblastoma
Optimization of Antibodies for Detection of the Mismatch Repair Proteins MLH1, MSH2, MSH6, and PMS2 Using a Biotin-Free Visualization System
2006-01-01
Optimization of Antibodies for Detection of the Mismatch Repair Proteins MLH1, MSH2, MSH6, and PMS2 Using a Biotin-Free Visualization System
2006-01-01
Monoclonal Antibody Therapy in Treating Patients With Primary or Metastatic Melanoma or Brain Tumors
Brain and Central Nervous System Tumors; Melanoma (Skin); Metastatic Cancer
Brain and Central Nervous System Tumors
1982-10-02
The usefulness of antibodies directed against tumour products for localisation and therapy of cancer is limited by the large proportion of administered antibody which remains non-specifically in the circulation and extravascular space. Liposomally entrapped second antibody (LESA) directed against the first (antitumour) antibody has been used to accelerate clearance of non-tumour-bound first antibody without affecting its clearance from the tumour. Intravenously administered LESA binds the first antibody and LESA is cleared by the reticuloendothelial system. LESA accelerated clearance of radiolabelled (I 131) antibody directed against carcinoembryonic antigen (CEA) from the circulation in four of five patients with gastrointestinal cancer, enhancing gamma-camera imaging of the tumour in three of them. These results suggest that LESA can improve the sensitivity and specificity of tumour imaging with radiolabelled antitumour antibody. This strategy may also have the potential to improve the therapeutic ratio of some toxic agents linked to antitumour antibody.
2006-01-01
Objective: To explore the roles played by autoantibodies in systemic lupus erythematosus. Methods: Serum anti-dsDNA antibody (with RIA) and serum anti-nucleosome antibody (AnuA), AHA, anti SmD1-, anti Ro60UD-, anti U1 -RNP- , anti-Ro52KD, anti-SSB antibodies (with anti-nucleo antibodies linear spectrum blotting method) were detected in 50 patients with clinically proven systemic lupus erythematosus. Results: The positive rate with anti-SmD1 antibody was highest (82%), followed by anti-Ro60KD antibody (80%) and AnuA (72%). Positive rate with anti dsDNA-, AHA, anti-U1-RNP-, anti- Ro52KD and anti SSB-antibodies was 44%, 32%, 58%, 48% and 24% respectively. Positive rate with anti-SC1-70, ACA and Jo-1 antibodies was extremely low (below 10%). Conclusion: Multiple auto-antibodies were present in serum of patients with systemic lupus erythematosus and combined ... >>
Sustained systemic delivery of monoclonal antibodies by genetically modified skin fibroblasts.
2000-01-01
In vivo production and systemic delivery of therapeutic antibodies by engineered cells might advantageously replace injection of purified antibodies for treating a variety of life-threatening diseases, including cancer, acquired immunodeficiency syndrome, and autoimmune diseases. We report here that skin fibroblasts retrovirally transduced to express immunoglobulin genes can be used for sustained long-term systemic delivery of cloned antibodies in immunocompetent mice. Importantly, no anti- idiotypic response against the ectopically expressed model antibody used in this study was observed. This supports the notion that skin fibroblasts can potentially be used in antibody-based gene/cell therapy protocols without inducing any adverse immune response in treated individuals.
Sustained systemic delivery of monoclonal antibodies by genetically modified skin fibroblasts.
2000-01-01
In vivo production and systemic delivery of therapeutic antibodies by engineered cells might advantageously replace injection of purified antibodies for treating a variety of life-threatening diseases, including cancer, acquired immunodeficiency syndrome, and autoimmune diseases. We report here that skin fibroblasts retrovirally transduced to express immunoglobulin genes can be used for sustained long-term systemic delivery of cloned antibodies in immunocompetent mice. Importantly, no anti- idiotypic response against the ectopically expressed model antibody used in this study was observed. This supports the notion that skin fibroblasts can potentially be used in antibody-based gene/cell therapy protocols without inducing any adverse immune response in treated individuals.
Sustained systemic delivery of monoclonal antibodies by genetically modified skin fibroblasts.
2000-01-01
In vivo production and systemic delivery of therapeutic antibodies by engineered cells might advantageously replace injection of purified antibodies for treating a variety of life-threatening diseases, including cancer, acquired immunodeficiency syndrome, and autoimmune diseases. We report here that skin fibroblasts retrovirally transduced to express immunoglobulin genes can be used for sustained long-term systemic delivery of cloned antibodies in immunocompetent mice. Importantly, no anti- idiotypic response against the ectopically expressed model antibody used in this study was observed. This supports the notion that skin fibroblasts can potentially be used in antibody-based gene/cell therapy protocols without inducing any adverse immune response in treated individuals.
Sustained systemic delivery of monoclonal antibodies by genetically modified skin fibroblasts.
2000-01-01
In vivo production and systemic delivery of therapeutic antibodies by engineered cells might advantageously replace injection of purified antibodies for treating a variety of life-threatening diseases, including cancer, acquired immunodeficiency syndrome, and autoimmune diseases. We report here that skin fibroblasts retrovirally transduced to express immunoglobulin genes can be used for sustained long-term systemic delivery of cloned antibodies in immunocompetent mice. Importantly, no anti- idiotypic response against the ectopically expressed model antibody used in this study was observed. This supports the notion that skin fibroblasts can potentially be used in antibody-based gene/cell therapy protocols without inducing any adverse immune response in treated individuals.
Sustained systemic delivery of monoclonal antibodies by genetically modified skin fibroblasts.
2000-01-01
In vivo production and systemic delivery of therapeutic antibodies by engineered cells might advantageously replace injection of purified antibodies for treating a variety of life-threatening diseases, including cancer, acquired immunodeficiency syndrome, and autoimmune diseases. We report here that skin fibroblasts retrovirally transduced to express immunoglobulin genes can be used for sustained long-term systemic delivery of cloned antibodies in immunocompetent mice. Importantly, no anti- idiotypic response against the ectopically expressed model antibody used in this study was observed. This supports the notion that skin fibroblasts can potentially be used in antibody-based gene/cell therapy protocols without inducing any adverse immune response in treated individuals.
Sustained systemic delivery of monoclonal antibodies by genetically modified skin fibroblasts.
2000-01-01
In vivo production and systemic delivery of therapeutic antibodies by engineered cells might advantageously replace injection of purified antibodies for treating a variety of life-threatening diseases, including cancer, acquired immunodeficiency syndrome, and autoimmune diseases. We report here that skin fibroblasts retrovirally transduced to express immunoglobulin genes can be used for sustained long-term systemic delivery of cloned antibodies in immunocompetent mice. Importantly, no anti- idiotypic response against the ectopically expressed model antibody used in this study was observed. This supports the notion that skin fibroblasts can potentially be used in antibody-based gene/cell therapy protocols without inducing any adverse immune response in treated individuals.
Suppression of the immune response to ovalbumin in vivo by anti-idiotypic antibodies
Conditions of suppression of the immune response to a food allergin (ovalbumin) were studied with the aid of anti-idiotypic (AID) antibodies. Hen ovalbumin was used and the experiments were performed on mice. Antibodies were isolated from the resulting protein fractions and tested for inhibitor activity by the method of direct radioimmunologic analysis. The test system consisted of the reaction of binding the globulin fraction to the total preparation of antibodies to ovalbumin from mice and a /sup 125/I-labeled total preparation of antibodies to ovalbumin of the same animals.
Suppression of the immune response to ovalbumin in vivo by anti-idiotypic antibodies
1986-12-01
Conditions of suppression of the immune response to a food allergin (ovalbumin) were studied with the aid of anti-idiotypic (AID) antibodies. Hen ovalbumin was used and the experiments were performed on mice. Antibodies were isolated from the resulting protein fractions and tested for inhibitor activity by the method of direct radioimmunologic analysis. The test system consisted of the reaction of binding the globulin fraction to the total preparation of antibodies to ovalbumin from mice and a /sup 125/I-labeled total preparation of antibodies to ovalbumin of the same animals.
1977-05-01
A solid-phase radioimmunoassay for the detection of anti-ribosome antibodies is described. Only small quantities of ribosomal particles are required, and low levels of antibody can be detected by very simple procedures. This assay has been used to detect antibodies which bind to intact ribosomes and to isolated large (60S) and small (40S) ribosomal subunits. The antibodies were present in sera from patients with systemic lupus erythematosus (SLE) and reacted predominantly with the 60S subunits. The potential use of the assay for the study of ribosome structure is discussed.
Radioimmunoassay for thyroid-stimulating hormone (TSH)
1978-05-09
This invention provides a method for the radioimmunoassay of thyroid-stimulating hormone which utilizes a rapid and convenient version of a double antibody procedure. Highly purified second antibody is bound, by means of covalent bonds, to hydrolyzed polyacrylamide particles to produce a two-phase system. The solid phase comprises immobilized second antibody bound to the reaction product of labeled and unlabeled thyroid-stimulating hormone with the first antibody (first antibody-antigen complex) and the liquid phase comprises free (unbound) labeled and unlabeled thyroid-stimulating hormone. The two phases are separated and the radioactivity of either phase is measured.
The effects of total lymphoid irradiation (TLI) on serum levels of autoantibodies, and of antibodies to diphtheria toxoid, tetanus toxoid, and pneumococcal polysaccharide in patients with lupus nephritis were compared with those previously observed in rheumatoid arthritis (RA) patients. Baseline levels of antibodies to diphtheria toxoid and tetanus toxoid decreased significantly after TLI in patients with lupus and RA, but antibody levels to pneumococcal polysaccharide remained unchanged. After TLI, the levels of antinuclear and anti-DNA antibodies were reduced significantly in lupus, but levels of rheumatoid factor, antinuclear, and antigranulocyte antibodies all tended to increase in RA.
1986-01-01
The effects of total lymphoid irradiation (TLI) on serum levels of autoantibodies, and of antibodies to diphtheria toxoid, tetanus toxoid, and pneumococcal polysaccharide in patients with lupus nephritis were compared with those previously observed in rheumatoid arthritis (RA) patients. Baseline levels of antibodies to diphtheria toxoid and tetanus toxoid decreased significantly after TLI in patients with lupus and RA, but antibody levels to pneumococcal polysaccharide remained unchanged. After TLI, the levels of antinuclear and anti-DNA antibodies were reduced significantly in lupus, but levels of rheumatoid factor, antinuclear, and antigranulocyte antibodies all tended to increase in RA.
1986-01-01
The effects of total lymphoid irradiation (TLI) on serum levels of autoantibodies, and of antibodies to diphtheria toxoid, tetanus toxoid, and pneumococcal polysaccharide in patients with lupus nephritis were compared with those previously observed in rheumatoid arthritis (RA) patients. Baseline levels of antibodies to diphtheria toxoid and tetanus toxoid decreased significantly after TLI in patients with lupus and RA, but antibody levels to pneumococcal polysaccharide remained unchanged. After TLI, the levels of antinuclear and anti-DNA antibodies were reduced significantly in lupus, but levels of rheumatoid factor, antinuclear, and antigranulocyte antibodies all tended to increase in RA
2003-01-01
Radioimmunoassay (RIA) is one of the in vitro diagnostic methods in nuclear medicine. The most important factor in RIA reagents to be considered is the antibody production, as specific antibodies with high affinity are the backbone of RIA techniques. In this experiment iodine (I125) radiolabelled antigens were used to evaluate locally produced donkey anti-sheep serum (DASS) as a separating agent in RIA. Two local donkeys were immunized with non-immunized sheep immunoglobulin (IgG) to produce donkey anti-sheep secondary antibodies. Samples were collected from the two donkeys, purified, dialysed, qualitatively tested for the presence of antibodies, which were then quantitatively evaluated and utilized as precipitating agent in RIA by adding the primary antibodies. Using RIA methodology the antibodies were titrated against three different analytes ... >>
Preparation and use of therapeutic antibodies primarily of human origin
2008-01-01
Therapeutic antibodies include polyclonal immunoglobulins isolated from regular or high-titered human plasma, sera from immunized animals, and monoclonal antibodies. This array of therapeutic antibodies is used for the prevention and treatment of many infectious diseases, antibody immunodeficiencies, autoimmune and inflammatory diseases, neurological disorders, and cancers. Polyclonal human immunoglobulins are available for intramuscular injection (IGIM), intravenous infusion (IGIV) and subcutaneous infusion (SCIG). We review these products and detail the therapeutic use of polyclonal human antibodies in the treatment of antibody immunodeficiencies, including their occasional local side effects (tenderness, sterile abscesses), minor systemic side effects (chills, muscle aches, malaise, hea...
2006-01-01
Maternal antibody prevents the use of live, attenuated measles vaccine (LAV) before 6–9 months of age, but vaccinated 6-month-old infants can mount a T cell response. An infant macaque model was used to study the immune response to LAV in the newborn in the presence or absence of maternal antibody. Four newborn monkeys without detectable maternal antibody and 9 newborns with passive measles antibody were vaccinated with LAV. Only the infants without passive antibody seroconverted after vaccination and 3 of 4 of these infants also developed measles-specific interferon γ+ T cells. The monkeys were challenged with wild-type measles virus at 5 months of age, and 7 of 9 infants vaccinated in the presence of passive antibody had systemic infection and skin rash, while 3 of the 4 infants vacci...
Identification and prevention of antibody disulfide bond reduction during cell culture manufacturing
2010-01-01
In the biopharmaceutical industry, therapeutic monoclonal antibodies are primarily produced in mammalian cell culture systems. During the scale-up of a monoclonal antibody production process, we observed excessive mechanical cell shear as well as significant reduction of the antibodys interchain disulfide bonds during harvest operations. This antibody reduction event was catastrophic as the product failed to meet the drug substance specifications and the bulk product was lost. Subsequent laboratory studies have demonstrated that cells subjected to mechanical shear release cellular enzymes that contribute to this antibody reduction phenomenon (manuscript submitted; Kao et al., 2009). Several methods to prevent this antibody reduction event were developed using a lab-scale model to reproduce...
Cancer imaging with radiolabeled antibodies
1990-01-01
This book presents a perspective of the use of antibodies to target diagnostic isotopes to tumors. Antibodies with reasonable specificity can be developed against almost any substance. If selective targeting to cancer cells can be achieved, the prospects for a selective therapy are equally intriguing. But the development of cancer detection, or imaging, with radiolabeled antibodies has depended upon advances in a number of different areas, including cancer immunology and immunochemistry for identifying suitable antigen targets and antibodies to these targets, tumor biology for model systems, radiochemistry for he attachment of radionuclides to antibodies, molecular biology for reengineering the antibodies for safer and more effective use in humans, and nuclear medicine for providing the best imaging protocols and instrumentation to detect minute amounts of elevated radioactivity against a background of considerable noise. Accordingly, this book has been organized to address the advances that are being made in many of these areas.
Radioimmunological determination of Fenoterol. Pt. 1. Theoretical fundamentals
1985-01-01
A general solution is given for the system of equations describing coupled mass equilibria having m antibodies with one binding site, or with several mutually independent binding sites of equal intrinsic affinity (immunoglobulin (Ig) G antibodies) and n monovalent antigens. Based on this a formula is given with which it is possible to calculate the minimum association constant of the antibody and the minimum specific radioactivity of the tracer in a radioimmunoassay when the detection limit is given and the assay conditions are established. The model m=2 and n=4 describes the behaviour of a system which is based on a mixture of stereoselective antibodies and a racemic tracer. The effect of the enantiomer ratio of an optically active ligand on this system is demonstrated.
A solid-phase immunosorbent radioassay for the detection of circulating antibodies to protein hormones is described. The assay is based on the binding of the homologous /sup 125/I-labelled antigen to the antibodies which are then bound to anti-IgG antibodies covalently coupled to Sepharose. It can easily be applied as a complement to any radioimmunoassay for the detection of circulating antibodies to the ligand measured. The assay system avoids falsely elevated values due to interference of high serum concentrations of the antigen. The assay was applied to measure antibodies to FSH, LH, TSH, GH, prolactin, insulin and thyroglobulin (Tg). Among patients with chronic thyroiditis Tg antibodies were found in 100% of the sera. In diffuse toxic goitre 73% of the patients had detectable Tg antibodies. Insulin antibodies were present in 82% of the sera from patients with insulin treated diabetes. No antibodies were found against the other protein hormones tested.
1984-10-01
A solid-phase immunosorbent radioassay for the detection of circulating antibodies to protein hormones is described. The assay is based on the binding of the homologous /sup 125/I-labelled antigen to the antibodies which are then bound to anti-IgG antibodies covalently coupled to Sepharose. It can easily be applied as a complement to any radioimmunoassay for the detection of circulating antibodies to the ligand measured. The assay system avoids falsely elevated values due to interference of high serum concentrations of the antigen. The assay was applied to measure antibodies to FSH, LH, TSH, GH, prolactin, insulin and thyroglobulin (Tg). Among patients with chronic thyroiditis Tg antibodies were found in 100% of the sera. In diffuse toxic goitre 73% of the patients had detectable Tg antibodies. Insulin antibodies were present in 82% of the sera from patients with insulin treated diabetes. No antibodies were found against the other protein hormones tested.
Monoclonal Antibodies as Diagnostics; an Appraisal
2010-01-01
Full Text Available.Ever since the development of Hybridoma Technology in 1975 by Kohler and Milstein, our vision for antibodies as tools for research for prevention, detection and treatment of diseases, vaccine production, antigenic characterization of pathogens and in the study of genetic regulation of immune responses and disease susceptibility has been revolutionized. The monoclonal antibodies being directed against single epitopes are homogeneous, highly specific and can be produced in unlimited quantities. In animal disease diagnosis, they are very useful for identification and antigenic characterization of pathogens. Monoclonal antibodies have tremendous applications in the field of diagnostics, therapeutics and targeted drug delivery systems, not only for infectious diseases caused by bacteria, viruses and protozoa but also for cancer, metabolic and hormonal disorders. They are also used in the diagnosis of lymphoid and myeloid malignancies, tissue typing, enzyme linked immunosorbent assay, radio immunoassay, serotyping of microorganisms, immunological intervention with passive antibody, antiidiotype inhibition, or magic bullet therapy with cytotoxic agents coupled with anti mouse specific antibody. Recombinant deoxyribonucleic acid technology through genetic engineering has successfully led to the possibility of reconstruction of monoclonal antibodies viz. chimeric antibodies, humanized antibodies and complementarily determining region grafted antibodies and their enormous therapeutic use.
Libraries against libraries for combinatorial selection of replicating antigen–antibody pairs
2009-02-03
Full Text Available.Antibodies are among the most highly selective tight-binding ligands for proteins. Because the human genome project has deciphered the proteome, there is an opportunity to use combinatorial antibody libraries to select high-affinity antibodies to every protein encoded by the genome. However, this is a large task because the selection formats used today for combinatorial antibody libraries are geared toward generating antibodies to one antigen at a time. Here, we describe a method that accelerates the identification of antibodies to a multitude of antigens simultaneously by matching combinatorial antibody libraries against eukaryotic antigen libraries so that replication-competent cognate antigen–antibody pairs can be directly selected. Phage and yeast display systems are used because they each link genotype to phenotype and can be replicated individually. When combined with cell sorting, the two libraries can be selected against each other for recovery of cognate antigen–antibody clones in a single experiment.
Chorea in systemic lupus erythematosus: association with antiphospholipid antibodies.
1988-08-01
Full Text Available.Chorea is a rare manifestation of systemic lupus erythematosus (SLE). In this report the clinical features of two cases of chorea associated with SLE are presented. Of special interest were the raised titres of antiphospholipid antibodies in both cases. The possible pathogenic role of these antibodies is briefly discussed.
Modulation of regulatory mechanisms operative in the cyclical production of antibody
1976-01-01
The response in rabbits to a single intravenous injection of aggregated human gamma globulin (AHuIgG) has been characterized by the successive appearance of peaks of plaque-forming cells (PFC) in the spleen and peripheral blood, separated by well-defined 8-day periods of decreased responsiveness. Although there is evidence in other systems for regulation of the immune response by circulating antibody, resulting in cyclical variations of antibody and PFC, the 8-day latent period in rabbit spleens is observed regardless of the dose of antigen injected or the amount of circulating antibody formed. Thus, it was postulated that if antibody played a role in this system, the antibody produced locally, at the site of PFC production, was the best candidate for exerting such precise control of the response.
Modulation of regulatory mechanisms operative in the cyclical production of antibody
1976-01-01
The response in rabbits to a single intravenous injection of aggregated human gamma globulin (AHuIgG) has been characterized by the successive appearance of peaks of plaque-forming cells (PFC) in the spleen and peripheral blood, separated by well-defined 8-day periods of decreased responsiveness. Although there is evidence in other systems for regulation of the immune response by circulating antibody, resulting in cyclical variations of antibody and PFC, the 8-day latent period in rabbit spleens is observed regardless of the dose of antigen injected or the amount of circulating antibody formed. Thus, it was postulated that if antibody played a role in this system, the antibody produced locally, at the site of PFC production, was the best candidate for exerting such precise control of the response
2008-01-01
Enhanced cell death and deficient clearance of cellular debris are thought to contribute to increased self-antigen exposure in systemic autoimmune disease. To investigate the characteristics of early humoral autoimmune responses, six monoclonal antibodies were generated from two autoimmune prone strains of mice. All antibodies specifically bound the surface of late-stage apoptotic cells. Similar antibody reactivities were present in the sera of patients with systemic lupus erythematosus. While IgM antibodies significantly reduced the phagocytic uptake of apoptotic thymocytes, IgG antibodies enhanced uptake. Poly-reactivity was demonstrated in the recognition of ribonucleoproteins and lipids. An antibody reactive towards lysophosphatidylcholine reversed lysophosphatidylcholine-mediated inhi...
Autoantibodies in the autoimmune rheumatic diseases
2010-01-01
The detection of autoantibodies to a variety of antigenic targets plays an important role in the diagnosis of autoimmune rheumatic disease. Rheumatoid factor, antinuclear antibodies and antineutrophil cytoplasmic antibodies have been the key in the diagnosis of rheumatoid arthritis (RA), systemic lupus erythematosus and systemic vasculitis for many years. Antibodies against cyclic citrullinated peptides have been shown to be specific for rheumatoid arthritis and may predict the development of erosive disease, and are being increasingly used in the assessment of patients with RA. There is increasing interest in the use of anti-C1q antibodies for the detection and monitoring of lupus nephritis as this assay appears to be more sensitive than anti-DNA antibodies. Antibodies against b2-glycopro...
An antibody profile of systemic lupus erythematosus detected by antigen microarray
2010-01-01
Summary Patients with systemic lupus erythematosus (SLE) produce antibodies to many different self-antigens. Here, we investigated antibodies in SLE sera using an antigen microarray containing many hundreds of antigens, mostly self-antigens. The aim was to detect sets of antibody reactivities characteristic of SLE patients in each of various clinical states - SLE patients with acute lupus nephritis, SLE patients in renal remission, and SLE patients who had never had renal involvement. The analysis produced two novel findings: (i) an SLE antibody profile persists independently of disease activity and despite long-term clinical remission, and (ii) this SLE antibody profile includes increases in four specific immunoglobulin G (IgG) reactivities to double-stranded DNA (dsDNA), single-stranded ...
1994-01-01
A human tumor xenograft model was used to compare the tumor and normal tissue uptake of a tumor-associated monoclonal antibody radiolabeled with 125I or 90Y. Nude mice bearing SC xenografts of the human colon adenocarcinoma, HT29, were injected with a mixture of 125I- and 90Y-DTPA-labeled AUA1 monoclonal antibody, which recognizes an antigen expressed on the surface of the tumor cells. In addition, the effect of systemic ethylenediaminetetraacetic acid (EDTA) administration on 90Y-labeled antibody clearance, tumor uptake of antibody and bone accumulation of 90Y was studied in a nude mouse model of intraperitoneal cancer. Both the absolute amount (%idcentre dotg-1) and the tumor:normal tissue ratios were superior for the 90Y-labeled antibody, compared ... >>
1992-06-01
Full Text Available.A competitive enzyme immunoassay (CEIA) was established and compared with other serological techniques for detecting Coxiella burnetii antibody in camels, goats, and sheep. This technique was evaluated because a conjugated anti-camel immunoglobulin was not available to serve as a direct signal for the demonstration of antigen-antibody reaction. A C. burnetii antibody-positive human serum and a peroxidase-conjugated anti-human immunoglobulin G were used as an indicator system competing against antibody in animal serum or as an indicator of the absence of antibody. Sera were considered antibody positive when the A414 of the test sera plus the competing positive antibody was less than or equal to 50% of the A414 of the negative-control serum plus the competing antibody. Antibody to C. burnetii was repeatedly demonstrated in 66% of camel serum samples (n = 200) by the CEIA. Among 48 camel serum samples, 71% were positive for antibody by CEIA versus 65% by EIA using peroxidase-labeled protein A. The CEIA detected C. burnetii antibody in 63% of sheep serum samples (n = 40) and in 50% of goat serum samples (n = 96), while the indirect fluorescent-antibody technique detected antibody in 38% of sheep and 34% of goat serum samples and the EIA detected antibody in 50% of sheep and 35% of goat serum samples. These data indicate that the CEIA is a reliable and sensitive technique for demonstrating C. burnetii antibody in camels, sheep, and goats.
2007-01-01
Transfusion medicine is an important part of modern health care and the provision of reliably phenotyped red blood cells (RBC) is essential for safe and effective blood transfusions. For identification of many RBC antigens, monoclonal antibodies of either murine or human origin are available for use in agglutination assays, in which they perform as well as or better than the human polyclonal antibody preparations which they have replaced. However, the detection of some blood groups is still reliant on the use of human polyclonal antisera, which is a less reliable reagent source with respect to availability, batch to batch variation and bio-safety. The use of recombinant antibody and phage display technology for the discovery of new monoclonal antibodies with specificity for some of these RBC antigens has the potential to deliver an economical, unlimited supply of specific antibody reagents suitable for use in RBC phenotyping. Samples of human B cells from donors producing useful phenotyping antibodies were identified and transformed using Epstein Barr virus into lymphocyte cell lines. Antibody genes were obtained from the cell lines in the form ofRNA which was reverse transcribed, amplified by PCR and cloned into a phagemid vector system to generate several combinatorial antibody libraries. These antibody libraries were displayed on the surface of phage particles and subjected to antigen-driven selection by several rounds of phage display biopanning using soluble and cell based RBC antigens. In addition a large naIve library was biopanned against the same antigens in an attempt to isolate a wide range of antibodies suitable for blood typing. Several high quality combinatorial antibody libraries with respect to size (> 107 clones) and diversity were generated. Biopanning of recombinant libraries resulted in enrichment of phage antibodies specific for RBC antigens, and several clones were isolated which were shown to be specific for Duffy a antigen. The isolated antibodies would be ideal candidates for re-engineering into multivalent antibody molecules capable of direct agglutination of RBC and as such, have the potential to replace human polyclonal sera in the identification of Duffy a RBC antigen phenotyping.
1991-12-01
To develop a homologous radioimmunoassay (RIA) for a hormone of a small or rare animal often meets difficulty in collecting a large amount of purified antigen required for antibody production. On the other hand, to employ a heterologous RIA to estimate the hormone often gives poor sensitivity. To overcome this difficulty, a 'hetero-antibody' RIA was studied. In a hetero-antibody RIA system, a purified preparation of a hormone is used for radioiodination and standardization and a heterologous antibody to the hormone is used for the first antibody. Canine motilin and rat LH were selected as examples, and anti-porcine motilin and anti-hCG, anti-hCG{beta} or anti-ovine LH{beta} was used as the heterologous antibody. The sensitivities of the hetero-antibody RIAs were much higher than those of heterologous RIAs in any case, showing that these hetero-antibody RIA systems were suitable for practical use. To clarify the principle of hetero-antibody RIA, antiserum to porcine motilin was fractionated on an affinity column where canine motilin was immobilized. The fraction bound had greater constants of affinity with both porcine and canine motilins than the rest of the antibody fractions. This fraction also reacted with a synthetic peptide corresponding to the C-terminal sequence common to porcine and canine motilins in a competitive binding test with labeled canine motilin. These results suggest that an antibody population having high affinity and cross-reactivity is present in polyclonal antiserum and indicate that the population can be used in hetero-antibody RIA at an appropriate concentration. (author).
Full Text Available.BackgroundMaternal antibodies are believed to play an integral role in protecting immunologically immature wild-passerines from environmental antigens. This study comprehensively examines the early development of the adaptive immune system in an altricial-developing wild passerine species, the house sparrow (Passer domestics), by characterizing the half-life of maternal antibodies in nestling plasma, the onset of de novo synthesis of endogenous antibodies by nestlings, and the timing of immunological independence, where nestlings rely entirely on their own antibodies for immunologic protection.Methodology/Principal FindingsIn an aviary study we vaccinated females against a novel antigen that these birds would not otherwise encounter in their natural environment, and measured both antigen-specific and total antibody concentration in the plasma of females, yolks, and nestlings. We traced the transfer of maternal antibodies from females to nestlings through the yolk and measured catabolisation of maternal antigen-specific antibodies in nestlings during early development. By utilizing measurements of non-specific and specific antibody levels in nestling plasma we were able to calculate the half-life of maternal antibodies in nestling plasma and the time point at which nestling were capable of synthesizing antibodies themselves. Based on the short half-life of maternal antibodies, the rapid production of endogenous antibodies by nestlings and the relatively low transfer of maternal antibodies to nestlings, our findings suggest that altricial-developing sparrows achieve immunologic independence much earlier than precocial birds.Conclusions/SignificanceTo our knowledge, this is the first in depth analyses performed on the adaptive immune system of a wild-passerine species. Our results suggest that maternal antibodies may not confer the immunologic protection or immune priming previously proposed in other passerine studies. Further research needs to be conducted on other altricial passerines to determine if the results of our study are a species-specific phenomenon or if they apply to all altricial-developing birds.
Target-specific binding of immunoliposomes in vivo
Our group at the University of Tennessee has been concentrating on using monoclonal antibody for targeting of a liposomal drug carrier system. This paper discusses our initial effort to target these liposomes using an organ-specific monoclonal antibody. 9 refs., 9 figs.
Target-specific binding of immunoliposomes in vivo
1989-01-01
Our group at the University of Tennessee has been concentrating on using monoclonal antibody for targeting of a liposomal drug carrier system. This paper discusses our initial effort to target these liposomes using an organ-specific monoclonal antibody. 9 refs., 9 figs.
Prevalence of Brucella abortus antibodies in serum of Holstein cattle in Cameroon
Holstein cattle of small scale dairy production systems were screened for Brucella abortus antibodies in 21 villages in Cameroon by ELISA. Results show a general seroprevalence of 8.4% in Holstein cat ... Full Text Available
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Neuromyelitis optica - an update: 2007–2009
2009-01-01
Neuromyelitis optica is an inflammatory demyelinating disorder of the central nervous system. The discovery of a specific antibody (NMO IgG /aquaporin-4 antibody) in patients with this condition has...Full Text Available
Immune molecules target swine- and avian-origin influenza
Immune molecules known as antibodies that protect against influenza virus infection target the highly variable influenza protein HA. It is thought the antibodies generated by an individual's immune system protect ...
Immune compound blocks virus' ability to hijack antibodies
Researchers at Washington University School of Medicine in St. Louis have shown that a controversial phenomenon known as antibody-dependent enhancement of infection is suppressed by C1q, a blood-borne immune system ...
IgM--IgG switch looked at from a control theoretic viewpoint. [Antigen-antibody reactions]
1977-01-01
The vertebrate immune system is described. Why a cell makes two different types of antibody with the same specificity for antigen is considered.
... Proper Name: Antibody to Hepatitis B Surface Antigen (Mouse Monoclonal) Enzyme-Linked Immunosorbent Assay (ELISA) (Antibody to HBsAG/Enzyme Immuno Assay (EIA)) ...
Caltech biologists spy on the secret inner life of a cell
The transportation of antibodies from a mother to her newborn child is vital for the development of that child's nascent immune system. Antibodies help shape a baby's response to foreign pathogens and may influence the ...
Biolex reports potential for more potent, efficacious antibodies in Nature Biotechnology
Biolex Therapeutics published results in Nature Biotechnology demonstrating ability of its proprietary LEX System to produce monoclonal antibodies with enhanced in vitro potency and efficacy. The research was conducted ...
Artificial immune system for optimal design of composite hydrogen storage vessel
2009-01-01
Based on the clonal selection principle in the biological immunology, an artificial immune system (AIS) is proposed to optimize the weight of Al-carbon fiber/epoxy composite hydrogen storage vessels under burst pressure constraint. The AIS simulates the generation of the antigens, the combination of the antigens and antibodies, and the selection, cloning and hypermutation of B cells, the maintenance of diverse antibody cells, the generation of the memory cells with high affinities and the death of the antibody cells with low affinities. Effects of the antibody size and the iterative number on the optimization results are explored. By comparison, the AIS shows higher search velocity and precision than the genetic algorithm and simulated annealing.
1995-02-01
Full Text Available.Experimental data suggest a role for the microflora in the disease expression of systemic lupus erythematosus (SLE). In active SLE anti-ds-DNA antibodies are supposed to be pathogenic by forming immune complexes with DNA. Bacteria might induce the production of anti-ds-DNA antibodies. To explore the relation between the host and his microflora in SLE in comparison with healthy controls we studied the prevalence of systemic antibodies to faecal bacteria that were discriminated by their morphology by indirect immunofluorescence. IgM titres against their own faecal microflora were found to be lower both in active and inactive SLE when compared to healthy individuals. IgG-class antibacterial antibodies were increased in inactive SLE but decreased in active SLE compared to inactive SLE and healthy controls, although plasma levels of total IgG were almost doubled in active SLE. The lower IgG antibacterial antibody titres in active SLE might possibly result from sequestration of these IgG antibodies in immune complexes, indicating a possible role for antibacterial antibodies in exacerbations of SLE.
1975-11-11
An apparatus and method for determining the type, amount and position of antibodies and/or antigens in a biological cell is described. Dyes which affect selected antibodies and/or antigens in a particular manner are introduced into a subject cell. An exemplary single channel system moves a light beam in an X-Y scan relative to the cell. The dyed antibodies and/or antigens, when excited by specific wavelengths in the light beam, respond by fluorescing at predetermined wavelengths or absorbing certain of the specific wavelengths. The particular wavelengths fluoresced or absorbed indicate the types of antibodies and/or antigens present in the cell. The system preferably outputs to a color television system to produce a display representative of the antibody and antigen distribution in the cell by type, amount, and position. The application also discloses multichannel embodiments.
1989-12-01
Full Text Available.Ten sera from 48 patients with systemic sclerosis were found to be capable of producing cytotoxicity of human umbilical venous and arterial endothelium when co-cultured with peripheral blood mononuclear cells. Fractionation of sera on Ultrogel and the preparation of monomeric IgG by ion exchange and affinity chromatography suggested that the cytotoxicity was mediated by anti-endothelial antibodies capable of pre-sensitizing target cells in a mechanism that resembled antibody-dependent cellular cytotoxicity. These anti-endothelial antibodies together with C1q-binding immune complexes and anti-cardiolipin antibodies were found in 18 of 28 patients so investigated, suggesting that multiple immunological mechanisms may be involved in the pathogenesis of the vascular lesion of systemic sclerosis.
Antibody Fab display system that can perform open-sandwich ELISA
2009-01-01
Previously, immunological detection of small haptens or peptides was only possible in a competitive format, which needed competitor antigen either labeled by a reporter or attached to a carrier protein. Beside this, open-sandwich immunoassay (OS-IA) is a simple but powerful immunoassay that can noncompetitively determine monovalent antigen concentration by measuring the antigen-dependent increase in VH/VL interaction of an antibody. However, the procedure to obtain suitable assay reagents for OS-IA for a target antigen has not been straightforward because of the lack of easy-to-use antibody selection/manipulation methods. Here we devise a new Fab antibody phage display system that is useful for rapidly evaluating and selecting suitable antibody Fv fragments to OS-IA. The system is based on...
Effect of electrolytes and of distilled water on antigen–antibody complexes
1971-11-01
Full Text Available. Specific immune precipitates dissolve in concentrated solutions of alkali-metal halides, and of alkaline-earth-metal halides and thiocyanates. The quantity of protein dissolved depends on the nature of the antigen–antibody system, on the proportion of the antigen in the precipitate, and on the avidity of the antibody. The extent of solubilization is a function of the temperature, of the volume of solution used and of the concentration of the ions in the solution, and also depends on the nature of these ions. The dissolving power of bivalent cations is greater than that of monovalent ones, and is as follows: Mg2+[unk]Ba2+[unk]Ca2+[unk]Sr2+. Antigen–antibody complexes and free antibodies, but no free antigen, are detected in supernatants of specific precipitates dissolved in solutions of electrolytes of low ionic strength. Antigen–antibody complexes, free antibodies and also free antigen are detected in supernatants of specific precipitates dissolved in solutions of electrolytes of high ionic strength. Comparable results are obtained when the electrolyte solutions are studied for their effect on the bonds formed between an antibody and its corresponding immunosorbent. Moreover, in the latter case, 50% of the fixed antibodies could be recovered by elution with distilled water.
The Chemistry of Irreversible Capture
2008-09-01
Full Text Available.The specific recognition and binding of biological molecules by antibodies is fundamentally important. Natural antibodies are multivalent, having at least two identical ligand-binding sites; this permits them to bind tightly at cell surfaces, which present multiple copies of their target ligands. Antibodies that bind to soluble monovalent ligands, such as most small molecules, do not share this multivalent advantage. Nor do engineered fragments of antibodies, such as single-chain Fv proteins or Fab fragments, which generally possess only a single ligand-binding site. Engineered monovalent antibody/ligand pairs that retain the binding specificity of the antibody, but do not dissociate, are promising components of new delivery systems. These are based on a combination of genetic manipulation of the protein and chemical synthesis of appropriate ligands, examples of which are reviewed here.
Production and Characterization of HCG Poly clonal Antibodies
2009-01-01
To prepare a radioimmunoassay system for measuring hcg hormone, the anti-hcg polyclonal antibodies must be firstly prepared. The present study was aimed to produce and characterize the anti-hcg polyclonal antibodies. The anti hcg polyclonal Antibes which were produced by immunizing five Balb/c mice with a highly purified beta-hcg hormone. A complete Freunds adjuvant was used for the first injection while incomplete Freunds adjuvant was used for the following boosters. Blood spots were obtained from the immunized mice and tested for the presence of antibodies. At the end of the immunization schedule, the mice was exposed to anesthesia using ether then dissected and the blood samples were collected from the heart. The serum (antisera) was separated from the blood containing anti-hcg polyclonal antibodies. The produced antibodies were purified then ... >>
The effect of total lymphoid irradiation (TLI) on the primary antibody response to the dinitrophenylated heterologous protein, keyhole limpet hemocyanin (DNP-KLH), in complete Freund's adjuvant (CFA), and to the trinitrophenylated polysaccharide antigen, Brucella abortus (TNP-BA), was studied in BALB/c mice. The antibody response to both antigens was diminished in comparison with nonirradiated mice when antigens were injected within 3 days after TLI. When the mice were immunized 30 days after completion of TLI the antibody response to DNP-KLH in CFA was still diminished, but the antibody response to TNP-BA was enhanced 5- to 10-fold as compared with that of control animals. The opposite effect of TLI on the two antibody responses was also observed in a syngeneic primary adoptive transfer system.
1984-02-01
The effect of total lymphoid irradiation (TLI) on the primary antibody response to the dinitrophenylated heterologous protein, keyhole limpet hemocyanin (DNP-KLH), in complete Freund's adjuvant (CFA), and to the trinitrophenylated polysaccharide antigen, Brucella abortus (TNP-BA), was studied in BALB/c mice. The antibody response to both antigens was diminished in comparison with nonirradiated mice when antigens were injected within 3 days after TLI. When the mice were immunized 30 days after completion of TLI the antibody response to DNP-KLH in CFA was still diminished, but the antibody response to TNP-BA was enhanced 5- to 10-fold as compared with that of control animals. The opposite effect of TLI on the two antibody responses was also observed in a syngeneic primary adoptive transfer system.
1984-01-01
The effect of total lymphoid irradiation (TLI) on the primary antibody response to the dinitrophenylated heterologous protein, keyhole limpet hemocyanin (DNP-KLH), in complete Freund's adjuvant (CFA), and to the trinitrophenylated polysaccharide antigen, Brucella abortus (TNP-BA), was studied in BALB/c mice. The antibody response to both antigens was diminished in comparison with nonirradiated mice when antigens were injected within 3 days after TLI. When the mice were immunized 30 days after completion of TLI the antibody response to DNP-KLH in CFA was still diminished, but the antibody response to TNP-BA was enhanced 5- to 10-fold as compared with that of control animals. The opposite effect of TLI on the two antibody responses was also observed in a syngeneic primary adoptive transfer system
1975-07-01
Using the adhering property of certain types of cells to glass or plastic in human peripheral blood lymphocyte preparations, two populations of effector cells can be identified in the ABCIL system, each with a selective killing directed toward a different antibody coated target. The adherent cells which are radioresistant functionally, with the morphology of macrophages, exhibit strong cytotoxicity against the antibody-coated erythroid target while lacking any killing effect toward the antibody-coated target lymphocytes. The nonadherent cells which are radiosensitive and with the morphology of small lymphocytes, selectively kill the antibody-coated lymphocyte target without possessing any cytotoxicity against the antibody-coated target erythrocytes.
1975-01-01
Using the adhering property of certain types of cells to glass or plastic in human peripheral blood lymphocyte preparations, two populations of effector cells can be identified in the ABCIL system, each with a selective killing directed toward a different antibody coated target. The adherent cells which are radioresistant functionally, with the morphology of macrophages, exhibit strong cytotoxicity against the antibody-coated erythroid target while lacking any killing effect toward the antibody-coated target lymphocytes. The nonadherent cells which are radiosensitive and with the morphology of small lymphocytes, selectively kill the antibody-coated lymphocyte target without possessing any cytotoxicity against the antibody-coated target erythrocytes
1975-01-01
Using the adhering property of certain types of cells to glass or plastic in human peripheral blood lymphocyte preparations, two populations of effector cells can be identified in the ABCIL system, each with a selective killing directed toward a different antibody coated target. The adherent cells which are radioresistant functionally, with the morphology of macrophages, exhibit strong cytotoxicity against the antibody-coated erythroid target while lacking any killing effect toward the antibody-coated target lymphocytes. The nonadherent cells which are radiosensitive and with the morphology of small lymphocytes, selectively kill the antibody-coated lymphocyte target without possessing any cytotoxicity against the antibody-coated target erythrocytes
2009-01-01
Live recombinant Saccharomyces cerevisiae yeast expressing the envelope antigen of Japanese encephalitis virus (JEV) on the outer mannoprotein layer of the cell wall were examined for their ability to induce antigen-specific antibody responses in mice. When used as a model antigen, parenteral immunization of mice with surface-expressing GFP yeast induced a strong anti-GFP antibody response in the absence of adjuvants. This antigen delivery approach was then used for a more stringent system, such as the envelope protein of JEV, which is a neurotropic virus requiring neutralizing antibodies for protection. Although 70% of cells were detected to express the total envelope protein on the surface by antibodies raised to the bacterially expressed protein, polyclonal anti-JEV antibodies failed to...
1974-01-01
A theory of antigen-antibody induced particulate aggregation is developed by investigating the stability of model systems of particles. A sufficient condition for the formation of large aggregate is derived by imposing the requirement that at equilibrium a statistically significant number of redundant bonds would occur in a reduced monomer-dimer model system. A basic relationship is obtained which predicts the fractional agglutination in the reduced dimer system as a function of the antigen, antibody and particulate concentrations.
1983-01-01
For MoAb to be used efficiently for drug targeting and tumor imaging, the fraction of antibody binding to tumor cells must be maximized. The authors have studied the binding of /sup 125/I MoAb in three different tumor systems. The fraction of antibody that could be bound to the cell surface was directly proportional to the antibody purity. The affinity constant also limits the fraction of antibody that can bind to cells at a given antigen concentration. Rearrangement of the standard expression for univalent equilibrium binding between two reactants shows that in antigen excess, the maximum fraction of antibody that can bind =Ka(Ag total)/1 + Ka(Ag total). Binding data using four different MoAb with three cell systems confirm this relationship. Estimates for reasonable concentrations of tumor antigens in vivo indicate that antibodies with binding constants less than 10/sup 8/ M/sup -1/ are not likely to be useful for drug targeting or tumor imaging.
1983-01-01
For MoAb to be used efficiently for drug targeting and tumor imaging, the fraction of antibody binding to tumor cells must be maximized. We have studied the binding of 125I MoAb in three different tumor systems. The fraction of antibody that could be bound to the cell surface was directly proportional to the antibody purity. The affinity constant also limits the fraction of antibody that can bind to cells at a given antigen concentration. Rearrangement of the standard expression for univalent equilibrium binding between two reactants shows that in antigen excess, the maximum fraction of antibody that can bind (formula; see text). Binding data using four different MoAb with three cell systems confirm this relationship. Estimates for reasonable concentrations of tumor antigens in vivo indicate that antibodies with binding constants less than 10(8) M-1 are not likely to be useful for drug targeting or tumor imaging.
1981-01-01
A solid-phase radioimmunoassay method was devised to detect mouse anti-double strand (ds) DNA antibody. This method could easily detect the anti-ds DNA antibody in 1 : 10,000 dilutions (1 unit) of pooled 9-10 month-old female NZB/W F1 sera. The sensitivity was about 103 and 102-fold higher than that of the modified Farr method and of the double antibody technique respectively. NZB/W mice developed high titer anti-dsDNA antibody as they grew older. Spleen cells brought to a microculture system using flat-bottomed polystyrene plates produced anti-dsDNA antibody clearly detectable by solid-phase radioimmunoassay. Anti-dsDNA antibody produced in vitro (y units) was in close correlation with the anti-dsDNA antibody titer of the spleen donor (x units) (y = 4.8 X 10-2 x-65, gamma = 0.94, P >
1987-01-01
The activation of oxygen metabolism in peritoneal macrophages during the defence against Francisella tularensis infection was inhibited by gamma irradiation of mice with 4.0 Gy. The application of specific antibodies protected the irradiated mice from the lethal infection without reactivation of oxygen metabolism in mononuclear phagocytes. These results demonstrated that the protecting function of the specific antibodies in the defence system against intracellular bacterial pathogens will be mediated by the oxygen-independent mechanisms. (author)
Technetium-99m imaging of melanoma with murine monoclonal antibodies
1989-12-01
This article on {sup 99m}Tc monoclonal antibody and its specific usage in the detection of melanoma is the first in a five-part update series. Upon completion of this article, the reader will understand: (1) the benefits of {sup 99m}Tc-labeled monoclonal antibodies; (2) the importance of careful labeling; and (3) the advantage of simultaneously imaging multiple organ systems.
Carboxypeptidase G2 (CP) is a bacterial enzyme, which is targeted to tumours by an antitumour antibody for local prodrug activation in antibody-directed enzyme prodrug therapy (ADEPT). Repeated cycles ... Full Text Available
Digital Repository Infrastructure Vision for European Research (DRIVER)
2009-02-01
Full Text Available.The sinopulmonary tract is the major site of infection in patients with primary antibody deficiency syndromes, and structural lung damage arising from repeated sepsis is a major determinant of morbidity and mortality. Patients with common variable immunodeficiency may, in addition, develop inflammatory lung disease, often associated with multi-system granulomatous disease. This review discusses the presentation and management of lung disease in patients with primary antibody deficiency.
2008-09-01
Full Text Available.Antibodies have proven to be effective agents in cancer imaging and therapy. One of the major challenges still facing the field is the heterogeneous distribution of these agents in tumors when administered systemically. Large regions of untargeted cells can therefore escape therapy and potentially select for more resistant cells. We present here a summary of theoretical and experimental approaches to analyze and improve antibody penetration in tumor tissue.
Anti-inflammatory activity of human IgA antibodies and their Fab alpha fragments: inhibition of IgG-mediated complement activation
1989-01-01
The interaction of human IgA antibodies with the classical pathway of complement activation was investigated in a homologous human system, by means of two IgA1 and three IgG1 myeloma proteins having antibody activity against a defined antigen, staphylococcal alpha-toxin. In a solid-phase antigen-dependent C3b-binding ELISA system, the monoclonal IgG antibodies were previously shown to activate the classical complement pathway synergistically, resembling polyclonal IgG antibodies, whereas IgA antibodies were unable to activate complement by either pathway. In the present study, IgA antibodies were found to inhibit significantly the activation of complement initiated by antigen-bound polyclonal or mixed monoclonal IgG antibodies, in relation to the amount of IgA antibodies applied and bound to antigen. IgA1 myeloma proteins devoid of antigen-binding activity were without effect. Inhibition was independent of the ability of the IgA antibodies to compete against the IgG antibodies in binding to antigen, and was demonstrable with physiological concentrations of antibodies. Similar results were obtained with polyclonal serum IgA having antigen-binding activity. However, the binding of C1q to antigen-complexed IgG was inhibited only by a monoclonal IgA antibody that could compete against one of the three monoclonal IgG antibodies that bound C1q synergistically. This observation implied that at least two mechanisms were involved in the inhibition of C3b fixation. Fab alpha fragments of monoclonal IgA antibodies, obtained by cleavage with IgA1 protease from Haemophilus influenzae type b, were found to have a similar inhibitory effect on C3b fixation to the intact IgA1 antibodies. This observation supports the hypothesis that IgA1 proteases contribute to the invasive pathogenicity of certain mucosal bacteria, by cleaving secretory IgA1 antibodies to antigen-binding Fab alpha fragments, which are not only defective in mucosal defense properties, but which also protect the organisms from other immune effector systems, such as complement activation.
Anti-inflammatory activity of human IgA antibodies and their Fab alpha fragments: inhibition of IgG-mediated complement activation
1989-01-01
The interaction of human IgA antibodies with the classical pathway of complement activation was investigated in a homologous human system, by means of two IgA1 and three IgG1 myeloma proteins having antibody activity against a defined antigen, staphylococcal alpha-toxin. In a solid-phase antigen-dependent C3b-binding ELISA system, the monoclonal IgG antibodies were previously shown to activate the classical complement pathway synergistically, resembling polyclonal IgG antibodies, whereas IgA antibodies were unable to activate complement by either pathway. In the present study, IgA antibodies were found to inhibit significantly the activation of complement initiated by antigen-bound polyclonal or mixed monoclonal IgG antibodies, in relation to the amount of IgA antibodies applied and bound to antigen. IgA1 myeloma proteins devoid of antigen-binding activity were without effect. Inhibition was independent of the ability of the IgA antibodies to compete against the IgG antibodies in binding to antigen, and was demonstrable with physiological concentrations of antibodies. Similar results were obtained with polyclonal serum IgA having antigen-binding activity. However, the binding of C1q to antigen-complexed IgG was inhibited only by a monoclonal IgA antibody that could compete against one of the three monoclonal IgG antibodies that bound C1q synergistically. This observation implied that at least two mechanisms were involved in the inhibition of C3b fixation. Fab alpha fragments of monoclonal IgA antibodies, obtained by cleavage with IgA1 protease from Haemophilus influenzae type b, were found to have a similar inhibitory effect on C3b fixation to the intact IgA1 antibodies. This observation supports the hypothesis that IgA1 proteases contribute to the invasive pathogenicity of certain mucosal bacteria, by cleaving secretory IgA1 antibodies to antigen-binding Fab alpha fragments, which are not only defective in mucosal defense properties, but which also protect the organisms from other immune effector systems, such as complement activation.
Lethal Antibody Enhancement of Dengue Disease in Mice Is Prevented by Fc Modification
2010-02-01
Full Text Available.Immunity to one of the four dengue virus (DV) serotypes can increase disease severity in humans upon subsequent infection with another DV serotype. Serotype cross-reactive antibodies facilitate DV infection of myeloid cells in vitro by promoting virus entry via Fcγ receptors (FcγR), a process known as antibody-dependent enhancement (ADE). However, despite decades of investigation, no in vivo model for antibody enhancement of dengue disease severity has been described. Analogous to human infants who receive anti-DV antibodies by transplacental transfer and develop severe dengue disease during primary infection, we show here that passive administration of anti-DV antibodies is sufficient to enhance DV infection and disease in mice using both mouse-adapted and clinical DV isolates. Antibody-enhanced lethal disease featured many of the hallmarks of severe dengue disease in humans, including thrombocytopenia, vascular leakage, elevated serum cytokine levels, and increased systemic viral burden in serum and tissue phagocytes. Passive transfer of a high dose of serotype-specific antibodies eliminated viremia, but lower doses of these antibodies or cross-reactive polyclonal or monoclonal antibodies all enhanced disease in vivo even when antibody levels were neutralizing in vitro. In contrast, a genetically engineered antibody variant (E60-N297Q) that cannot bind FcγR exhibited prophylactic and therapeutic efficacy against ADE-induced lethal challenge. These observations provide insight into the pathogenesis of antibody-enhanced dengue disease and identify a novel strategy for the design of therapeutic antibodies against dengue.
1968-06-01
Full Text Available. A group of sera from hospital patients and normal subjects has been tested for the presence of antinuclear antibodies, and the amount of labile inhibitor to desoxyribonuclease 1 present in each of the sera estimated. It has been found that the presence of antinuclear antibodies is associated with a high level of inhibitor to a statistically significant degree. It has also been found that this inhibitor is formed during the clotting of whole blood and that it is derived from platelets. A possible relationship between the inhibitor, its formation during the clotting of blood, and the characteristic presence of antinuclear antibodies in systemic lupus erythematosus is discussed.
Serological testing for trichinosis in pigs
A test system suitable for rapid and economical screening of large numbers of swine for antibodies to Trichinella spiralis is being developed. It was demonstrated that the soluble antigen-fluorescent antibody (SAFA) test can detect T. spiralis antibodies earlier and in swine fed fewer larvae than other tests. An enzyme-labeled antibody test (ELA) is being developed which is similar in principle to SAFA, but was shown to be more sensitive than SAFA by comparison in a double blind test on sera from experimentally infected pigs.
Serological testing for trichinosis in pigs
1976-01-01
A test system suitable for rapid and economical screening of large numbers of swine for antibodies to Trichinella spiralis is being developed. It was demonstrated that the soluble antigen-fluorescent antibody (SAFA) test can detect T. spiralis antibodies earlier and in swine fed fewer larvae than other tests. An enzyme-labeled antibody test (ELA) is being developed which is similar in principle to SAFA, but was shown to be more sensitive than SAFA by comparison in a double blind test on sera from experimentally infected pigs.
Properties, production, and applications of camelid single-domain antibody fragments
2007-11-01
Full Text Available.Camelids produce functional antibodies devoid of light chains of which the single N-terminal domain is fully capable of antigen binding. These single-domain antibody fragments (VHHs or Nanobodies®) have several advantages for biotechnological applications. They are well expressed in microorganisms and have a high stability and solubility. Furthermore, they are well suited for construction of larger molecules and selection systems such as phage, yeast, or ribosome display. This minireview offers an overview of (1) their properties as compared to conventional antibodies, (2) their production in microorganisms, with a focus on yeasts, and (3) their therapeutic applications.
Impact of bulking agents on the stability of a lyophilized monoclonal antibody
2009-01-01
The impact of the bulking agents on the stability of lyophilized protein pharmaceuticals is not typically considered, as they usually crystallize, preventing them from stabilizing the protein. In this study, combinations of sucrose with mannitol or glycine were evaluated for their effects on antibody stability (deamidation and aggregation) and solid-state properties (water content, residual antibody structure, and Tg values). While sucrose remains the primary stabilizing agent for the antibody in the solid state, inclusion of some amorphous glycine leads to a significant increase in stability. Mannitol displays little, if any, stabilizing ability in this system. In formulations where infrared spectroscopy could be applied, maintenance of secondary structure is an important factor in predic...
Specific immune responses but not basal functions of B and T cells are impaired in aged mice
2009-01-01
Senescence is characterized by several alterations in the immune system. Such modifications can be found in lymphoid organs as well as in the cellular components of the immune system. Several reports have suggested that immune dysfunction can affect both T and B cells, but T cells have been shown to be more susceptible to the effects of aging. B cell function may also be altered with reduction in germinal center formation, antibody response, and affinity maturation of antibodies. Herein we showed that although antigen-specific antibody response to a soluble antigen declines in 18-month old mice, total levels of serum antibodies as well as frequencies of spleen and bone marrow antibody-producing cells are increased in aged mice. In addition, proliferative response of non-stimulated spleen T...
Polyreactive autoantibodies in systemic lupus erythematosus have pathogenic potential
2009-01-01
The present study was undertaken to determine whether germline encoded and polyreactive antibodies might be pathogenic and whether the breach of early tolerance checkpoints in systemic lupus erythematosus (SLE) might lead to a population of B cells expressing germline encoded antibodies that become pathogenic merely by class switching to IgG in a pro-inflammatory milieu. We demonstrate here that IgM, DNA-reactive antibodies obtained from lupus patients that are unmutated and display polyreactivity can bind to isolated glomeruli and exhibit neurotoxic potential. Thus, the IgM polyreactive repertoire in SLE includes antibodies that may acquire pathogenic function merely by undergoing class-switch recombination to become IgG antibodies.
2005-01-01
Summary Background Monoclonal antibodies are a valuable tool in the study of allergens, but the technology used in their generation can be slow and labour-intensive. Therefore, we have examined recombinant antibody development by phage-display against single allergens and protein mixtures. Objective We used the avian immunoglobulin system (generated from single VH and VL genes) to provide a rapid method for generating highly specific recombinant antibody fragments from a minimal number of animals. Methods A single-chain antibody fragment (scFv) library was generated from a single chicken immunized with model allergens. ScFvs were isolated by phage-display and their properties investigated by ELISA and Western blot. Results Mono-specific scFvs were generated against recombinant Fel d 1 and ...
Autoantibodies as predictive tools in systemic sclerosis
2010-01-01
The pathogenetic role of autoantibodies in systemic sclerosis (SSc) remains unclear, but these autoantibodies have been established as strong predictors of disease outcome and the pattern of organ complications in patients with this condition. The three most frequently observed types of SSc-specific autoantibody—anti-centromere antibodies, anti-topoisomerase antibodies and anti-RNA polymerase III antibodies—are found in over 50% of patients; the presence of each is generally exclusive of the others. Although a lot less frequently observed, antibodies directed against U3RNP and Th/To are also specific for scleroderma, whereas anti-Pm/Scl, anti-Ku and anti-U1RNP antibodies are seen mainly in patients with overlap syndromes. Up to 11% of patients with SSc can test negative for ant...
Antibody engineering-a valuable asset in preventing closed environment epidemics
2005-01-01
Investigations of Mir, Space Shuttle, Skylab and Apollo missions report extensive colonisation of the spacecraft by bacteria and fungi, which can lead to degradative effects on spacecraft equipment and devastating effects on space-grown crops. More than 80% of terrestrial greenhouse epidemics are due to the fungal genera Phytophthora, Pythium and Fusarium, which have been found in life support system test-beds. The advent of recombinant antibody technologies, including ribosome display and phage display, has made it possible to develop antibodies against virtually any toxin or organism and allows for maturation of antibodies by in vitro molecular evolution. These antibodies may play an important role in an integrated pest management regime for life support systems. Efficacy of existing fun...
2009-01-01
Abstract Aim: To measure the level of anti-nucleosome antibodies in systemic lupus erythematosus (SLE) patients, to determine the sensitivity and the specificity of these antibodies in the diagnosis of the disease and to evaluate the relationship between the levels of anti-nucleosome antibodies, anti-dsDNA (double-stranded DNA) and SLE disease activity. Methods: A cross-sectional study was conducted. All patients attended either a medical specialist clinic or were admitted to the medical wards of Hospital Universiti Sains Malaysia with the diagnosis of SLE (n = 90), other connective tissue diseases (n = 45) or were normal controls (n = 90) within the period from July 2004 until September 2005. They were tested for anti-nucleosome antibodies by enzyme-linked immunosorbent assay and anti-DNA...
Anti-cyclic citrullinated peptide antibodies in juvenile idiopathic arthritis
2010-01-01
Objective Prevalence and clinical significance of anti-cyclic citrullinated peptide (CCP) antibodies in Indian patients with juvenile idiopathic arthritis (JIA). Methods Anti-CCP antibodies were determined by enzyme-linked immunosorbent assay (ELISA) in 78 patients with JIA which included all 3 major subtypes of the disease: pauciarticular, polyarticular afld systemic onset. Values above 5 relative units were taken as positive. Associations between antiCCP antibodies and clinical and laboratory and radiological parameters were determined. Results Anti-CCP antibodies were positive in only 2 of 34 (5.9%) patients with pauciarticular JIA and 3 of 17 (17.6%) of systemic,.pnset JIA, whereas it was positive in 13 of 27 (48.1%) of polyarticular JIA patients (p
Anti-C1q antibodies: association with nephritis and disease activity in systemic lupus erythematosus
2009-01-01
Background: Anti-C1q antibodies have been described in systemic lupus erythematosus (SLE) as well as in other connective tissue diseases. They have been considered as a marker for disease activity and presence of nephritis. Objective: The aim of this study was to determine the prevalence of anti-C1q antibodies in Brazilian lupus patients as well as analyze their association with different clinical and serologic parameters. Methods: Sera from 81 SLE patients, based on the American College of Rheumatology (ACR) criteria, were collected from a lupus referral outpatient clinic in Salvador, Brazil. Antibodies to C1q were detected by an enzyme-linked immunoassay (ELISA) kit and antibodies to other cellular antigens identified by indirect immunofluorescence on HEp-2 cell substrate (ANA), or Crith...
1976-01-01
32P-cyclophosphamide was found to combine with gamma-globulin fractions of immune sera. Immune sera incubated with 32P-cyclophosphamide retained ability to react specifically with homologouus antigen in vitro in the system: MN antigens of human erythrocytes + rabbit anti-MN antibody, and probably reacted selectively with target antigens in vivo in the system: antigens of guinea pig kidney tissue + rabbit antibodies against these antigens. Hemagglutination, passive hemagglutination and precipitation in agar gel tests were used in the experiments. Ability to combine of the immune antibody + 32P-cyclophosphamide complex with homologous antigens was evaluated by measurements of radioactivity of studied materials (erythrocyte agglutinates and organ homogenates). The results indicate feasibility of using ... >>
Cysticercosis of the nervous system. Treatment by means of specific internal radiation
Five hundred patients with cysticercosis of the nervous system were evaluated by scanning that used anti-Cysticercus antibodies labeled with indium 113. The same antibodies, labeled with iodine 131, were used for radioimmunotreatment. Ninety-six percent of the patients had good or excellent results, whereas only 4% had poor results. None of the patients showed intolerance or radiotoxicity during three months of clinical and laboratory follow-up. The diagnosis, treatment, and prognosis of cysticercosis of the nervous system are dramatically changing, due to the development of anti-Cysticercus antibodies labeled with radionuclides.
Cysticercosis of the nervous system. Treatment by means of specific internal radiation
Five hundred patients with cysticercosis of the nervous system were evaluated by scanning that used anti-Cysticercus antibodies labeled with indium 113. The same antibodies, labeled with iodine 131, were used for radioimmunotreatment. Ninety-six percent of the patients had good or excellent results, whereas only 4% had poor results. None of the patients showed intolerance or radiotoxicity during three months of clinical and laboratory follow-up. The diagnosis, treatment, and prognosis of cysticercosis of the nervous system are dramatically changing, due to the development of anti-Cysticercus antibodies labeled with radionuclides.
Cysticercosis of the nervous system. Treatment by means of specific internal radiation
1981-05-01
Five hundred patients with cysticercosis of the nervous system were evaluated by scanning that used anti-Cysticercus antibodies labeled with indium 113. The same antibodies, labeled with iodine 131, were used for radioimmunotreatment. Ninety-six percent of the patients had good or excellent results, whereas only 4% had poor results. None of the patients showed intolerance or radiotoxicity during three months of clinical and laboratory follow-up. The diagnosis, treatment, and prognosis of cysticercosis of the nervous system are dramatically changing, due to the development of anti-Cysticercus antibodies labeled with radionuclides.
Cysticercosis of the nervous system. Treatment by means of specific internal radiation
1981-05-01
Five hundred patients with cysticercosis of the nervous system were evaluated by scanning that used anti-Cysticercus antibodies labeled with indium 113. The same antibodies, labeled with iodine 131, were used for radioimmunotreatment. Ninety-six percent of the patients had good or excellent results, whereas only 4% had poor results. None of the patients showed intolerance or radiotoxicity during three months of clinical and laboratory follow-up. The diagnosis, treatment, and prognosis of cysticercosis of the nervous system are dramatically changing, due to the development of anti-Cysticercus antibodies labeled with radionuclides.
1988-10-01
Rats were injected both intradermally and intravenously with an IgG/sub 2/b mouse monoclonal antibody (791T/36) and subsequently the biodistribution of intravenously injected /sup 111/In labelled antibody was examined by gamma camera imaging in these and control rats. The majority of pretreated rats showed a marked perturbation of the biodistribution of the radiolabelled antibody with a marked increase of tracer in the liver. There was similar perturbation of the biodistribution of subsequently administered /sup 111/In labelled Fab fragment of the 791T/36 antibody and of another IgG/sub 2/b monoclonal antibody. In contrast, there was little influence on the biodistribution of IgG/sub 2/a or IgG/sub 1/ monoclonal antibodies, indicating that the response in the rats was predominantly against the IgG/sub 2/b isotype. This rat model system is amenable to examination of a number of aspects of the biological consequences of immune responses to foreign immunoglobulins relevant to their use for clinical imaging.
Rapid antibody selection by mRNA display on a microfluidic chip
2009-05-01
Full Text Available.In vitro antibody-display technologies are powerful approaches for isolating monoclonal antibodies from recombinant antibody libraries. However, these display techniques require several rounds of affinity selection which is time-consuming. Here, we combined mRNA display with a microfluidic system for in vitro selection and evolution of antibodies and achieved ultrahigh enrichment efficiency of 106- to 108-fold per round. After only one or two rounds of selection, antibodies with high affinity and specificity were obtained from naïve and randomized single-chain Fv libraries of ∼1012 molecules. Furthermore, we confirmed that not only protein–protein (antigen–antibody) interactions, but also protein–DNA and protein–drug interactions were selected with ultrahigh efficiencies. This method will facilitate high-throughput preparation of antibodies and identification of protein interactions in proteomic and therapeutic fields.
1982-02-01
The radioimmunoassay for gastrin is based on competition by unlabeled antigen, i.e., gastrin in the form of synthetic human gastrin I heptadecapeptide (SHG) as standard and (or) gastrin unknown with radiolabeled gastrin (/sub 125/I-SHG), for specific binding sites on antibodies to gastrin, produced in experimental animals. The concentrations of antibody and radiolabeled antigen are kept constant, and standard calibration curves are established by including various concentrations of unlabeled gastrin in the incubation tubes. The antigen, gastrin, is added to concentrations in excess of antibody, to ensure competition between /sub 125/I-SHG and gastrin for the available antibody-binding sites. In this system of competitive inhibition, the proportion of antibody-bound /sub 125/I-SHG is inversely related to the quantity of unlabeled gastrin included in the incubation mixture. Unlabeled antigen (gastrin) available for binding by antibody is either the gastrin present in the standards included in the construction of calibration curves or gastrin present in the unknown samples to be measured. As with other radioimmunoassay techniques, radioimmunoassay of gastrin requires: (a) production of antibodies specific for gastrin, (b) availability of a pure form of the peptide for radiolabeling, (c) preparation of a radiolabeled form of gastrin, and (d) an effective method of separating antibody-bound peptide from free.
1986-01-01
An approach to suppressing secondary disease with antibodies was studied that differed from conventional antibody treatment of donor marrow in vitro. It consisted of the selection of anti-Thy-1 antibodies with high affinity for Clq, the first subunit of the complement cascade, and a single injection of such antibodies into prospective irradiated marrow recipients. Monoclonal mouse IgM and rat IgG 2c antibodies of high titers in complement-dependent test systems but with low affinity for Clq caused little immunosuppression. Monoclonal rat IgG2b or mouse IgG2a anti-Thy-1 antibodies with high affinity for Clq prevented acute and chronic mortality of graft-v-host disease (GVHD), however, when injected in irradiated CBA or AKR mice prior to C57BL/6 spleen and/or bone marrow cell transfusion. This treatment simultaneously suppressed residual host-v-graft reactivity of ... >>
Antibody Maturation in Trypanosoma cruzi-Infected Rats
2001-07-01
Full Text Available.The study of antibody avidity changes during infection has improved the understanding of the pathologic processes involved in several infectious diseases. In some infections, like toxoplasmosis, this information is being used for diagnostic purposes. Results of the evolution of antibody avidity for different specific antigens in Trypanosome cruzi-infected rats are presented. A Western blotting technique, combined with avidity analysis to identify antigens that elicit high-avidity antibodies, is suggested. In this system, antibodies showed high avidity values only during the chronic phase of infection and only in relation to antibodies against 21-, 33-, 41-, 42-, 56-, 58-, 66-, and 72-kDa antigens. Finally, a 97-kDa T. cruzi antigen, which was recognized by high-avidity antibodies and occurred in noninfected rats, was identified. These results allow us to evaluate the different antigens in chagasic infection. Our results show that with the correct choice of antigen it is possible to detect differences in maturation of antibodies and to discriminate, in an experimental model, between recent (acute) and chronic infections.
Effect of different hapten-carrier conjugation ratios and molecular orientations on antibody affinity against a peptide antigen
2006-01-01
Hapten-carrier conjugate immunization is an important tool in the generation of hapten-specific antibodies for analytical purposes and in the uncovering of basic vaccine-immunological mechanisms. The affinity of antibodies is known to play an important role for the resulting sensitivity of antibody-based assay systems and in deciding whether a vaccine-induced antibody response will be protective. With ovalbumin as a carrier protein and a peptide (7.2NY) representing a 19 ammo acid sequence from the E. coli-derived Verotoxin 2e as a model hapten we investigated whether it was possible to influence the affinity and titre of antibodies raised against the hapten using different conjugation ratios and orientations. The peptide was coupled to ovalbumin in four Conjugation ratios and two molecular orientations - terminal and central - and the Conjugates were verified by mass spectrometry. Mice were immunised ten dines at two-weeks intervals with low doses of the eight conjugates, Blood samples collected between each immunisation were analysed by ELISA for specific antibody titres and relative affinities. With both types of conjugations, the anti-peptide antibody titres increased in response to increasing conjugation ratios, but central conjugation resulted in markedly higher titres than terminal Conjugation. The overall anti-peptide antibody affinities reached approximately similar levels with both orientations. whereas a reversed proportionality was observed between conjugation ratio and antibody affinity for terminal conjugation. Thus, it appears that the molar ratio of a peptide and its carrier may affect the resulting antibody affinities, and that a conjugation ratio between a terminally Conjugated peptide and its carrier approaching one will result in relatively high antibody affinities. Furthermore, the molecular orientation of the Coupled peptide has a major effect on the anti-peptide antibody titres induced.
Effect of different hapten-carrier conjugation ratios and molecular orientations on antibody affinity against a peptide antigen
2006-01-01
Hapten-carrier conjugate immunization is an important tool in the generation of hapten-specific antibodies for analytical purposes and in the uncovering of basic vaccine-immunological mechanisms. The affinity of antibodies is known to play an important role for the resulting sensitivity of antibody-based assay systems and in deciding whether a vaccine-induced antibody response will be protective. With ovalbumin as a carrier protein and a peptide (7.2NY) representing a 19 ammo acid sequence from the E. coli-derived Verotoxin 2e as a model hapten we investigated whether it was possible to influence the affinity and titre of antibodies raised against the hapten using different conjugation ratios and orientations. The peptide was coupled to ovalbumin in four Conjugation ratios and two molecular orientations - terminal and central - and the Conjugates were verified by mass spectrometry. Mice were immunised ten dines at two-weeks intervals with low doses of the eight conjugates, Blood samples collected between each immunisation were analysed by ELISA for specific antibody titres and relative affinities. With both types of conjugations, the anti-peptide antibody titres increased in response to increasing conjugation ratios, but central conjugation resulted in markedly higher titres than terminal Conjugation. The overall anti-peptide antibody affinities reached approximately similar levels with both orientations. whereas a reversed proportionality was observed between conjugation ratio and antibody affinity for terminal conjugation. Thus, it appears that the molar ratio of a peptide and its carrier may affect the resulting antibody affinities, and that a conjugation ratio between a terminally Conjugated peptide and its carrier approaching one will result in relatively high antibody affinities. Furthermore, the molecular orientation of the Coupled peptide has a major effect on the anti-peptide antibody titres induced.
Effect of different hapten-carrier conjugation ratios and molecular orientations on antibody affinity against a peptide antigen
2006-01-01
Hapten-carrier conjugate immunization is an important tool in the generation of hapten-specific antibodies for analytical purposes and in the uncovering of basic vaccine-immunological mechanisms. The affinity of antibodies is known to play an important role for the resulting sensitivity of antibody-based assay systems and in deciding whether a vaccine-induced antibody response will be protective. With ovalbumin as a carrier protein and a peptide (7.2NY) representing a 19 ammo acid sequence from the E. coli-derived Verotoxin 2e as a model hapten we investigated whether it was possible to influence the affinity and titre of antibodies raised against the hapten using different conjugation ratios and orientations. The peptide was coupled to ovalbumin in four Conjugation ratios and two molecular orientations - terminal and central - and the Conjugates were verified by mass spectrometry. Mice were immunised ten dines at two-weeks intervals with low doses of the eight conjugates, Blood samples collected between each immunisation were analysed by ELISA for specific antibody titres and relative affinities. With both types of conjugations, the anti-peptide antibody titres increased in response to increasing conjugation ratios, but central conjugation resulted in markedly higher titres than terminal Conjugation. The overall anti-peptide antibody affinities reached approximately similar levels with both orientations. whereas a reversed proportionality was observed between conjugation ratio and antibody affinity for terminal conjugation. Thus, it appears that the molar ratio of a peptide and its carrier may affect the resulting antibody affinities, and that a conjugation ratio between a terminally Conjugated peptide and its carrier approaching one will result in relatively high antibody affinities. Furthermore, the molecular orientation of the Coupled peptide has a major effect on the anti-peptide antibody titres induced.
Effect of different hapten-carrier conjugation ratios and molecular orientations on antibody affinity against a peptide antigen
2006-01-01
Hapten-carrier conjugate immunization is an important tool in the generation of hapten-specific antibodies for analytical purposes and in the uncovering of basic vaccine-immunological mechanisms. The affinity of antibodies is known to play an important role for the resulting sensitivity of antibody-based assay systems and in deciding whether a vaccine-induced antibody response will be protective. With ovalbumin as a carrier protein and a peptide (7.2NY) representing a 19 ammo acid sequence from the E. coli-derived Verotoxin 2e as a model hapten we investigated whether it was possible to influence the affinity and titre of antibodies raised against the hapten using different conjugation ratios and orientations. The peptide was coupled to ovalbumin in four Conjugation ratios and two molecular orientations - terminal and central - and the Conjugates were verified by mass spectrometry. Mice were immunised ten dines at two-weeks intervals with low doses of the eight conjugates, Blood samples collected between each immunisation were analysed by ELISA for specific antibody titres and relative affinities. With both types of conjugations, the anti-peptide antibody titres increased in response to increasing conjugation ratios, but central conjugation resulted in markedly higher titres than terminal Conjugation. The overall anti-peptide antibody affinities reached approximately similar levels with both orientations. whereas a reversed proportionality was observed between conjugation ratio and antibody affinity for terminal conjugation. Thus, it appears that the molar ratio of a peptide and its carrier may affect the resulting antibody affinities, and that a conjugation ratio between a terminally Conjugated peptide and its carrier approaching one will result in relatively high antibody affinities. Furthermore, the molecular orientation of the Coupled peptide has a major effect on the anti-peptide antibody titres induced.
Effect of different hapten-carrier conjugation ratios and molecular orientations on antibody affinity against a peptide antigen
2006-01-01
Hapten-carrier conjugate immunization is an important tool in the generation of hapten-specific antibodies for analytical purposes and in the uncovering of basic vaccine-immunological mechanisms. The affinity of antibodies is known to play an important role for the resulting sensitivity of antibody-based assay systems and in deciding whether a vaccine-induced antibody response will be protective. With ovalbumin as a carrier protein and a peptide (7.2NY) representing a 19 ammo acid sequence from the E. coli-derived Verotoxin 2e as a model hapten we investigated whether it was possible to influence the affinity and titre of antibodies raised against the hapten using different conjugation ratios and orientations. The peptide was coupled to ovalbumin in four Conjugation ratios and two molecular orientations - terminal and central - and the Conjugates were verified by mass spectrometry. Mice were immunised ten dines at two-weeks intervals with low doses of the eight conjugates, Blood samples collected between each immunisation were analysed by ELISA for specific antibody titres and relative affinities. With both types of conjugations, the anti-peptide antibody titres increased in response to increasing conjugation ratios, but central conjugation resulted in markedly higher titres than terminal Conjugation. The overall anti-peptide antibody affinities reached approximately similar levels with both orientations. whereas a reversed proportionality was observed between conjugation ratio and antibody affinity for terminal conjugation. Thus, it appears that the molar ratio of a peptide and its carrier may affect the resulting antibody affinities, and that a conjugation ratio between a terminally Conjugated peptide and its carrier approaching one will result in relatively high antibody affinities. Furthermore, the molecular orientation of the Coupled peptide has a major effect on the anti-peptide antibody titres induced.
Effect of different hapten-carrier conjugation ratios and molecular orientations on antibody affinity against a peptide antigen
2006-01-01
Hapten-carrier conjugate immunization is an important tool in the generation of hapten-specific antibodies for analytical purposes and in the uncovering of basic vaccine-immunological mechanisms. The affinity of antibodies is known to play an important role for the resulting sensitivity of antibody-based assay systems and in deciding whether a vaccine-induced antibody response will be protective. With ovalbumin as a carrier protein and a peptide (7.2NY) representing a 19 ammo acid sequence from the E. coli-derived Verotoxin 2e as a model hapten we investigated whether it was possible to influence the affinity and titre of antibodies raised against the hapten using different conjugation ratios and orientations. The peptide was coupled to ovalbumin in four Conjugation ratios and two molecular orientations - terminal and central - and the Conjugates were verified by mass spectrometry. Mice were immunised ten dines at two-weeks intervals with low doses of the eight conjugates, Blood samples collected between each immunisation were analysed by ELISA for specific antibody titres and relative affinities. With both types of conjugations, the anti-peptide antibody titres increased in response to increasing conjugation ratios, but central conjugation resulted in markedly higher titres than terminal Conjugation. The overall anti-peptide antibody affinities reached approximately similar levels with both orientations. whereas a reversed proportionality was observed between conjugation ratio and antibody affinity for terminal conjugation. Thus, it appears that the molar ratio of a peptide and its carrier may affect the resulting antibody affinities, and that a conjugation ratio between a terminally Conjugated peptide and its carrier approaching one will result in relatively high antibody affinities. Furthermore, the molecular orientation of the Coupled peptide has a major effect on the anti-peptide antibody titres induced.
Effect of different hapten-carrier conjugation ratios and molecular orientations on antibody affinity against a peptide antigen
2006-01-01
Hapten-carrier conjugate immunization is an important tool in the generation of hapten-specific antibodies for analytical purposes and in the uncovering of basic vaccine-immunological mechanisms. The affinity of antibodies is known to play an important role for the resulting sensitivity of antibody-based assay systems and in deciding whether a vaccine-induced antibody response will be protective. With ovalbumin as a carrier protein and a peptide (7.2NY) representing a 19 ammo acid sequence from the E. coli-derived Verotoxin 2e as a model hapten we investigated whether it was possible to influence the affinity and titre of antibodies raised against the hapten using different conjugation ratios and orientations. The peptide was coupled to ovalbumin in four Conjugation ratios and two molecular orientations - terminal and central - and the Conjugates were verified by mass spectrometry. Mice were immunised ten dines at two-weeks intervals with low doses of the eight conjugates, Blood samples collected between each immunisation were analysed by ELISA for specific antibody titres and relative affinities. With both types of conjugations, the anti-peptide antibody titres increased in response to increasing conjugation ratios, but central conjugation resulted in markedly higher titres than terminal Conjugation. The overall anti-peptide antibody affinities reached approximately similar levels with both orientations. whereas a reversed proportionality was observed between conjugation ratio and antibody affinity for terminal conjugation. Thus, it appears that the molar ratio of a peptide and its carrier may affect the resulting antibody affinities, and that a conjugation ratio between a terminally Conjugated peptide and its carrier approaching one will result in relatively high antibody affinities. Furthermore, the molecular orientation of the Coupled peptide has a major effect on the anti-peptide antibody titres induced.
Effect of different hapten-carrier conjugation ratios and molecular orientations on antibody affinity against a peptide antigen
2006-01-01
Hapten-carrier conjugate immunization is an important tool in the generation of hapten-specific antibodies for analytical purposes and in the uncovering of basic vaccine-immunological mechanisms. The affinity of antibodies is known to play an important role for the resulting sensitivity of antibody-based assay systems and in deciding whether a vaccine-induced antibody response will be protective. With ovalbumin as a carrier protein and a peptide (7.2NY) representing a 19 ammo acid sequence from the E. coli-derived Verotoxin 2e as a model hapten we investigated whether it was possible to influence the affinity and titre of antibodies raised against the hapten using different conjugation ratios and orientations. The peptide was coupled to ovalbumin in four Conjugation ratios and two molecular orientations - terminal and central - and the Conjugates were verified by mass spectrometry. Mice were immunised ten dines at two-weeks intervals with low doses of the eight conjugates, Blood samples collected between each immunisation were analysed by ELISA for specific antibody titres and relative affinities. With both types of conjugations, the anti-peptide antibody titres increased in response to increasing conjugation ratios, but central conjugation resulted in markedly higher titres than terminal Conjugation. The overall anti-peptide antibody affinities reached approximately similar levels with both orientations. whereas a reversed proportionality was observed between conjugation ratio and antibody affinity for terminal conjugation. Thus, it appears that the molar ratio of a peptide and its carrier may affect the resulting antibody affinities, and that a conjugation ratio between a terminally Conjugated peptide and its carrier approaching one will result in relatively high antibody affinities. Furthermore, the molecular orientation of the Coupled peptide has a major effect on the anti-peptide antibody titres induced.
Effect of different hapten-carrier conjugation ratios and molecular orientations on antibody affinity against a peptide antigen
2006-01-01
Hapten-carrier conjugate immunization is an important tool in the generation of hapten-specific antibodies for analytical purposes and in the uncovering of basic vaccine-immunological mechanisms. The affinity of antibodies is known to play an important role for the resulting sensitivity of antibody-based assay systems and in deciding whether a vaccine-induced antibody response will be protective. With ovalbumin as a carrier protein and a peptide (7.2NY) representing a 19 ammo acid sequence from the E. coli-derived Verotoxin 2e as a model hapten we investigated whether it was possible to influence the affinity and titre of antibodies raised against the hapten using different conjugation ratios and orientations. The peptide was coupled to ovalbumin in four Conjugation ratios and two molecular orientations - terminal and central - and the Conjugates were verified by mass spectrometry. Mice were immunised ten dines at two-weeks intervals with low doses of the eight conjugates, Blood samples collected between each immunisation were analysed by ELISA for specific antibody titres and relative affinities. With both types of conjugations, the anti-peptide antibody titres increased in response to increasing conjugation ratios, but central conjugation resulted in markedly higher titres than terminal Conjugation. The overall anti-peptide antibody affinities reached approximately similar levels with both orientations. whereas a reversed proportionality was observed between conjugation ratio and antibody affinity for terminal conjugation. Thus, it appears that the molar ratio of a peptide and its carrier may affect the resulting antibody affinities, and that a conjugation ratio between a terminally Conjugated peptide and its carrier approaching one will result in relatively high antibody affinities. Furthermore, the molecular orientation of the Coupled peptide has a major effect on the anti-peptide antibody titres induced.
Effect of different hapten-carrier conjugation ratios and molecular orientations on antibody affinity against a peptide antigen
2006-01-01
Hapten-carrier conjugate immunization is an important tool in the generation of hapten-specific antibodies for analytical purposes and in the uncovering of basic vaccine-immunological mechanisms. The affinity of antibodies is known to play an important role for the resulting sensitivity of antibody-based assay systems and in deciding whether a vaccine-induced antibody response will be protective. With ovalbumin as a carrier protein and a peptide (7.2NY) representing a 19 ammo acid sequence from the E. coli-derived Verotoxin 2e as a model hapten we investigated whether it was possible to influence the affinity and titre of antibodies raised against the hapten using different conjugation ratios and orientations. The peptide was coupled to ovalbumin in four Conjugation ratios and two molecular orientations - terminal and central - and the Conjugates were verified by mass spectrometry. Mice were immunised ten dines at two-weeks intervals with low doses of the eight conjugates, Blood samples collected between each immunisation were analysed by ELISA for specific antibody titres and relative affinities. With both types of conjugations, the anti-peptide antibody titres increased in response to increasing conjugation ratios, but central conjugation resulted in markedly higher titres than terminal Conjugation. The overall anti-peptide antibody affinities reached approximately similar levels with both orientations. whereas a reversed proportionality was observed between conjugation ratio and antibody affinity for terminal conjugation. Thus, it appears that the molar ratio of a peptide and its carrier may affect the resulting antibody affinities, and that a conjugation ratio between a terminally Conjugated peptide and its carrier approaching one will result in relatively high antibody affinities. Furthermore, the molecular orientation of the Coupled peptide has a major effect on the anti-peptide antibody titres induced.
Use of Genetic Systems HIV-2 EIA
... antibodies was estimated to be 99.8% in United States blood and plasma establishments ... which is widespread in the world, including a major epidemic focus in the ...
This document describes a protocol for qualitative determination of total complement activation by Western blot. Complement system represents an innate arm of immune defense and is named so because it ?complements? the antibody-mediated immune response.
Solid phase radioimmunoassay for thyroxine hormone (Preprint No. CT-64)
1988-01-01
Solid phase radioimmunoassay system for measurement of serum thyroxine was developed using polystyrene beads coated with thyroxine antibodies. (author)
2006-01-01
This article presents a real-time monitoring system for cellular analysis using micro total analysis systems technology. Time-resolved luminescence anisotropy analysis was adopted for real-time detection of small amounts of a target protein produced by a small number of cells. The system was tested by real-time monitoring of the antibody secretion by hybridomas. The cells were successfully cultivated in a micro-incubation chamber (240nl) fabricated on a microchip. The quantification of the antibody was achieved using the Ru(II) complex-labeled Staphylococcus aureus protein A probe, which can bind specifically to the Fc region of the antibody. Using this system, we detected as little as 24fmol of immunoglobulin G under physiological conditions without the bound/free separation protocol. We ...
PMA - Isolex 300 Magnetic Cell Selection System -Hematopoietic ...
... Isolex@ Stem Cell Reagent Kit A reagent kit, containing the biological reagents for ... particulates in the antibody product, and clumping and/or poor yield in the ...
Monoclonal antibodies directed against surface molecules of multicell spheroids
... a good model system to study the interactions of mammalian cells, and thereby provide a functional assay for surface adhesion molecules. This project also involves investigations of ...
Label-free Nanowire SARS Virus Detection System
Antibody mimic protein is tailored to attach to nanowire base at one end, leaving biologically active area open for detection.
Guidance for Industry: Donor Screening for Antibodies to HTLV-II
... I was identified as the etiologic agent of a number of disorders, including adult T ... may also be obtained by mail by calling the CBER Voice Information System at ...
Guidance for Industry: Availability of Licensed Donor Screening ...
... Systems Corporation's Supplement to its Product License Application for Antibody to Hepatitis B Surface Antigen (Mouse ... Contact Us. ... Food and Drug Administration. ...
Growing B Lymphocytes in a Three-Dimensional Culture System
... candidates in vitro, generation of human monoclonal antibodies, and immunotherapy. Mature lymphocytes, which are the effectors of adaptive immune responses in vertebrates, are ...
Growing B Lymphocytes in a Three-Dimensional Culture System
... candidates in vitro, generation of human monoclonal antibodies, and immunotherapy. Mature lymphocytes, which are the effectors of adaptive immune responses in vertebrates, ...
Growing B Lymphocytes in a Three-Dimensional Culture System
... candidates in vitro, generation of human monoclonal antibodies, and immunotherapy. Mature lymphocytes, which are the effectors of adaptive immune responses in vertebrates, are ...
Evidence of SV40 infections in hospitalized children
... and was significantly associated with kidney transplants (6 of 15 [40] positive, P < .001). Many of the antibody-positive patients had impaired immune systems. Molecular assays ...
Development of technics and reagents for assessment of viral ecology Final report
Feasibility of antigen/antibody system utilizing passive immune agglutination technique to determine microbial ecology of crew during extended space flight
... Ability to calculate interpreted results of cell panels and antibody identification; ... Operating System: Servers, Microsoft Windows XP, Microsoft Windows NT version ...
Antibodies linked to cardiovascular disease increase in patients with active lupus
A study by researchers in Australia and the United Kingdom suggests that autoantibodies to fat binding proteins significantly increase in systemic lupus erythematosus patients with active disease. Complete findings of ...
Anti-GD2 3F8 Antibody and Allogeneic Natural Killer Cells for High-Risk Neuroblastoma
Neuroblastoma; Bone Marrow, Sympathetic Nervous System
A Humanized Anti-Interferon-γ Monoclonal Antibody (HuZAF) for Moderate to Severe Crohn's Disease
Crohn's Disease; Colitis; Intestinal Disease; Gastrointestinal Disease; Digestive System Disease
Modulation of Antibody-Mediated Immune Response by Probiotics in Chickens
2005-12-01
Full Text Available.Probiotic bacteria, including Lactobacillus acidophilus and Bifidobacterium bifidum, have been shown to enhance antibody responses in mammals. The objective of this study was to examine the effects of a probiotic product containing the above bacteria in addition to Streptococcus faecalis on the induction of the chicken antibody response to various antigens, both systemically and in the gut. The birds received probiotics via oral gavage and subsequently were immunized with sheep red blood cells (SRBC) and bovine serum albumin (BSA) to evaluate antibody responses in serum or with tetanus toxoid (TT) to measure the mucosal antibody response in gut contents. Control groups received phosphate-buffered saline. Overall, BSA and SRBC induced a detectable antibody response as early as week 1 postimmunization (p.i.), which lasted until week 3 p.i. Probiotic-treated birds had significantly (P ≤ 0.001) more serum antibody (predominantly immunoglobulin M [IgM]) to SRBC than the birds that were not treated with probiotics. However, treatment with probiotics did not enhance the serum IgM and IgG antibody responses to BSA. Immunization with TT resulted in the presence of specific IgA and IgG antibody responses in the gut. Again, treatment with probiotics did not change the level or duration of the antibody response in the gut. In conclusion, probiotics enhance the systemic antibody response to some antigens in chickens, but it remains to be seen whether probiotics have an effect on the generation of the mucosal antibody response.
In Vitro Antibody-Enzyme Conjugates with Specific Bactericidal Activity
1973-06-01
Full Text Available.IgG with antibacterial antibody opsonic activity was isolated from rabbit antisera produced by intravenous hyperimmunization with several test strains of pneumococci, Group A β-hemolytic streptococci, Staphylococcus aureus, Proteus mirabilis, Pseudomonas aeruginosa, and Escherichia coli. Antibody-enzyme conjugates were prepared, using diethylmalonimidate to couple glucose oxidase to IgG antibacterial antibody preparations. Opsonic human IgG obtained from serum of patients with subacute bacterial endocarditis was also conjugated to glucose oxidase. Antibody-enzyme conjugates retained combining specificity for test bacteria as demonstrated by indirect immunofluorescence. In vitro test for bactericidal activity of antibody-enzyme conjugates utilized potassium iodide, lactoperoxidase, and glucose as cofactors. Under these conditions glucose oxidase conjugated to antibody generates hydrogen peroxide, and lactoperoxidase enzyme catalyzes the reduction of hydrogen peroxide with simultaneous oxidation of I- and halogenation and killing of test bacteria. Potent in vitro bactericidal activity of this system was repeatedly demonstrated for antibody-enzyme conjugates against pneumococci, streptococci, S. aureus, P. mirabilis, and E. coli. However, no bactericidal effect was demonstrable with antibody-enzyme conjugates and two test strains of P. aeruginosa. Bactericidal activity of antibody-enzyme conjugates appeared to parallel original opsonic potency of unconjugated IgG preparations. Antibody-enzyme conjugates at concentrations as low as 0.01 mg/ml were capable of intense bactericidal activity producing substantial drops in surviving bacterial counts within 30-60 min after initiation of assay. These in vitro bactericidal systems indicate that the concept of antibacterial antibody-enzyme conjugates may possibly be adaptable as a mechanism for treatment of patients with leukocyte dysfunction or fulminant bacteremia.
RedSoar--a system for red blood cell antibody identification.
1991-01-01
Full Text Available.The primary goal of our research is to build an intelligent tutoring system for red blood cell antibody identification. In this paper, we describe the basis for a tutoring system--an expert system called RedSoar. RedSoar is built from two task-specific architectures that were designed for building flexible systems (i.e. systems that can use a variety of problem-solving strategies, and to which knowledge can easily-be added). RedSoar solves the antibody identification task correctly 81% of the time and new knowledge can be added in a straightforward manner. The system is capable of exhibiting human-like behavior which we believe is a necessary condition for building a successful tutoring system.
2009-01-01
Background Anti-cyclic citrullinated peptide (anti-CCP) antibodies have proved to be a specific marker for the diagnosis of rheumatoid arthritis (RA). However, the antibodies can also be detected in other rheumatic diseases, especially systemic lupus erythematosus (SLE). Recent studies have shown anti-CCP antibodies are associated with erosive arthritis in SLE patients. Since erosive arthritis is not common in SLE and many patients with non-erosive arthritis also have anti-CCP antibodies, the clinical significance of anti-CCP antibodies in SLE needs to be further studied. Objective To investigate the prevalence and clinical significance of anti-CCP antibodies in Chinese SLE patients. Methods Serum samples from 138 SLE patients were examined for anti-CCP with the second generation anti-CCP ...
2009-01-01
The dual-vector system-II (DVS-II), which allows efficient display of Fab antibodies on phage, has been reported previously, but its practical applicability in a phage-displayed antibody library has not been verified. To resolve this issue, we created two small combinatorial human Fab antibody libraries using the DVS-II, and isolation of target-specific antibodies was attempted. Biopanning of one antibody library, termed DVFAB-1L library, which has a 1.3 Ã 107 combinatorial antibody complexity, against fluorescein-BSA resulted in successful isolation of human Fab clones specific for the antigen despite the presence of only a single light chain in the library. By using the unique feature of the DVS-II, an antibody library of a larger size, named DVFAB-131L, which has a 1.5 Ã 109 combinato...
1987-05-01
Antibodies against a cell-surface protein, cross-reactive with double-stranded DNA, were detected in the serum of 25 patients with active human systemic lupus erythematosus (SLE), defined on the basis of the revised American Rheumatism Association classification. Among these sera, two did not display anti-DNA antibodies, as shown by Farr assay, solid-phase radioimmunoassay, and Crithidia luciliae test. Five other SLE patients were consecutively studied in active and remission states. Antibodies against the protein were detected in the serum of the 5 SLE patients when they were in active phase but not in the serum of the same patients in inactive phase of the disease. The anti-protein antibodies were not found in the serum of 10 inactive SLE patients or in the sera of 10 normal human controls, 10 patients with rheumatoid arthritis, 5 patients with scleroderma, and 4 patients with primary sicca syndrome. Taken together, these results strongly suggest that antibodies against this cell-surface protein could provide a better diagnosis marker and activity index than anti-DNA antibodies in SLE.
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