Development of an ELISA for paraquat; improvement of antibody characteristics by reversed affinity chromatography.
نویسندگان
چکیده
An enzyme-linked immunosorbent assay (ELISA) for paraquat is described. The microtitration plate-based assay has a limit of detection of 10 pg per well, and was specific for paraquat and monoquat. Reversed affinity chromatography was used to refine the polyclonal antibody preparation and eliminate interfering antibodies. There were consequent and significant improvements in assay sensitivity and performance. The potential for application of the assay to a variety of matrices is discussed.
منابع مشابه
Measurement of Affinity Constant of Anti-human IgG Monoclonal Antibodies by an ELISA-based Method
Background: The affinity of an antibody to its antigen is a crucial parameter in its biological activity and performance of an immunoassay such as ELISA. Affinity of most IgG specific MAbs are often determined by methods which require labeling of either antigen or antibody, and are sometimes difficult to control, do not always lead to the expected signal and often result in immunological modifi...
متن کاملPurification of human anti-erythropoietin polyclonal antibodies by precipitation and chromatography as an optimized method with potential application in vaccine studies
Introduction: Polyclonal antibodies are required to be affinity purified. Improved purification methods of polyclonal antibody provide an opportunity to pick the most purified immunoglobulins as a primary or secondary antibody in immunoassays that are included in many vaccine studies. Two common techniques for purifying proteins is salt precipitation and chromatography purification. Our work fo...
متن کاملتولید انبوه آنتیبادی مونوکلونال علیه پپتید خارج سلولی CD20 و ارزیابی اتصال اختصاصی آن به سلولهای بیانکننده CD20
Background & Aims: Nowadays, by advent of hybridoma technology in monoclonal antibodies production various cell markers could be evaluated in malignant and non-malignant cells. CD20, non-glycosylated phosphoprotein is as an ideal marker in leukemia and B-cell lymphoma diagnosis which is expressed on more than 95% of normal and neoplastic B-cells except for early B-cells and mature plasma. Th...
متن کاملIsolation and purification of HLA-DR antigen from Daudi cell line by immunoaffinity chromatography
Introduction: The major histocompatibility complex (MHC) is a group of cell surface proteins that are essential for recognizing foreign molecules in human and other mammals. The physiologic function of MHC molecules is the presentation of peptides to T cells. In this study, we evaluated the purification of a class II MHC molecule (HLA-DR) from a human Burkitt′s lymphoma cell line; Daudi...
متن کاملPolyclonal Antibody Production against Mouse Purified IgG2a towards Use in Basic Research
Background: The ability of polyclonal antibodies to react with many epitopes of an antigen makes them valuable reagents in research and diagnosis. The aim of this study was purification of mouse IgG2a and production of polyclonal antibody against purified mouse IgG2a subclass. Materials and Methods: Mouse IgG2a was purified by ProA affinity. Verification method of the purified antibody was S...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- The Analyst
دوره 124 6 شماره
صفحات -
تاریخ انتشار 1999