Polyclonal antibodies antigen

The definition of an association constant for an antibody—antigen reaction can become more complex if the antibody—antigen reaction iavolves a multivalent antigen, as is the case when a polyclonal antisemm is used for detection of an antigen. This type of multivalent binding is termed avidity and is defined by the equation ... 

When an antigen is injected into an animal, the resulting antibodies are polyclonal, being synthesized by a mixture of B cells. Polyclonal antibodies are directed against a number of different sites (epitopes or determinants) on the antigen and thus are not monospecific. However, by means of a method developed by Kohler and Milstein, large amounts of a single monoclonal antibody specific for one epitope can be obtained. 

Shi S-R, Cote RJ, Liu C, et al. A modified reduced temperature antigen retrieval protocol effective for use with a polyclonal antibody to cyclooxygenase-2 (PG 27). Appl. Immunohistochem. Mol. Morphol. 2002 10 368-373. 

Mighell AJ, Robinson PA, Hume WJ. Patterns of immunoreactivity to an anti-fibronectin polyclonal antibody in formalin-fixed, paraffin-embedded oral tissues are dependent on methods of antigen retrieval. J. Histochem. Cytochem. 1995 43 1107-1114. 

Many variations on the assay exist, but the ELISA, shown schematically below, is currently highly favored because of its simplicity once established in a laboratory sensitivity, detecting about one adduct per 107 bases and ability to screen many samples because of easy automations. The current prerequisite for the assay is that DNA can be modified to sufficiently high levels with the ultimate carcinogen to make it suitably antigenic. These types of antigens have been used to raise polyclonal antibodies in rabbits (41) and monoclonal antibodies from mice (42). 

Immunoassays employ monoclonal or polyclonal antibody preparations (Chapter 13) to detect and quantify the product (Box 7.1). The specificity of antibody-antigen interaction ensures good assay precision. The use of conjugated radiolabels (RIA) or enzymes (EIA) to allow detection of antigen-antibody binding renders such assays very sensitive. Furthermore, when compared with... 

Immunoaffinity chromatography (IAC), 6 400—402 12 137, 145 Immunoanalyzers, automated, 14 150 Immunoassay(s), 14 135-159. See also Immunoassay- DNA probe hybrid assays Immunoassay methods Immuno(bio)sensors antibody-antigen reaction, 14 136-138 basic technology in, 14 138-140 chemiluminescent, 14 150-151 classification of, 14 140-153 design of, 14 139-140 enzyme, 14 143-148 fluorescence, 14 148-150 highly specific, 14 153 historical perspective on, 14 136 microarrays and, 14 156—157 microfluidics in, 26 968—969 monoclonal versus polyclonal antibodies in, 14 152-153... 

Antiserum (plural antisera) Blood serum containing one or more polyclonal antibodies that are specific for one or more antigens. The antibodies in an antiserum result from previous immunization or exposure to an agent of disease. 

Polyclonal antibodies are multiple antibodies produced by different types of immune cells (different clones) that recognize the same antigen, whereas monoclonal antibodies are derived from a single cell line. 

The binding of the antigen and antibody can be affected by several factors, including the conjugated label chosen for detection and the method used to conjugate the label, as well as the assay format itself. The selectivity of the ELISA can be affected by the assay format. In an ELISA with a two-site sandwich format, independent epitopes are bound by different antibodies.26 The specificity comes from multiple site recognition. Polyclonal antibodies can react with many epitopes on a complex antigen surface.24... 

Although polyclonal antibodies have been used for passive immunization and therapeutic treatments, there are cases when hypersensitivities are induced. The reason is that polyclonal antibodies contain not only the specific antibody that binds to the desired antigen but other antibodies, which our immune system will treat as foreign substances and act against. 

Polyclonal antibodies are produced by injecting an antigen into an animal in the presence of an adjuvant containing bacterial lipopolysaccharides that stimulate the immune system. Serum prepared from the blood contains several different classes of antibodies that interact with different domains in the antigen molecule, each of... 

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