Antibodies heterogeneity

The problem of antibody heterogeneity may well be complicating the precise delineation of antigenic determinants of proteins. Thus far all efforts to locate the antigenic determinants have been made with heterogeneous antibody populations. The subject must be reexplored with antiprotein antibodies obtained from hybridomas, which would provide monoclonal antibody populations. In this way one cannot have uniform reactivity with only the antigen itself, but must have it also with the vari-... 

Individual animals immunized with the same substance can produce antibodies that may differ in affinities, titer, and specificities. " Such differences are appeu-ent with antibodies studied by the more classical physical chemical procedures. However, even among anti-enzyme sera harvested from individual rabbits that had been immunized with -lactamase, some were found to neutralize the activity of the enzyme, others to stimulate its activity, and still others were stimulatory and then inhibitory at higher concentrations." In radioimmunoassay, each antiserum from an individual animal must be characterized separately to select those that have the proper affinities and specificities. The production of monoclonal antibodies by hybridoma technology can yield molecules with defined specificities and affinities. As this technology becomes affordable for the average research laboratory, problems associated with antibody heterogeneity are due to diminish. 

Polyclonal antibodies Heterogeneous antibody populations that exhibit different properties and binding characteristics Precision Extent to which certain measurements agree to each other, usually stated as coefficients of variation, standard deviation or confidence limits... 

Dionex Corporation (2009) Dionex Application Note No. 127 Monitoring Monoclonal Antibody Heterogeneity by Cation Exchnge Chromatography. Dionex Corporation (now part of Thermo Fisher Scientific), Sunnyvale, CA, USA. 

Definition of the association (or avidity) constant for such multivalent antibody—antigen reactions must consider not only the heterogeneity of the antibodies and the antigen determinant site(s), but also an apparent additive effect of binding two antigen molecules to a single antibody. Such effects lead... 

Most immunoassay kits and many commercial immunoassay analyzers are based on heterogenous EIA or FIA. These include an immunoassay system that uses FIA linked to radial partition chromatography of the antibody—antigen complex (39) a system that uses antibody-coated tubes for enzyme immunoassay of a variety of hormones and dmgs (40) and a system that uses either a sandwich or competitive FIA format to measure a variety of analytes (41). 

Nisonoff, A., and Pressman, D. Heterogeneity and average combining constants of antibodies from individual rabbits. J. Immunol 80 417-428, 1958. 

The DELFIA assay, the first effective lanthanide-based immunoassay, was developed and commercialized by the early 1980s.108-112 DELFIA (Dissociation Enhanced Lanthanide Fluoro-ImmunoAssay) is a heterogeneous assay which uses a lanthanide complex based on aminocarboxylate ligands such as EDTA, EGTA, or DTPA, linked to the antibody by reaction of appended isothiocyanate groups (e.g., complex (45)) with nucleophilic residues, particularly amines, on the protein surface (Figure 11). 

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