Analysis of anti-insulin antibodies using time-resolved fluoroimmunoassay.
This paper describes the analysis of both monoclonal and polyclonal murine insulin antibodies with time-resolved fluoroimmunoassay (TRFIA). This assay is based on the binding of antiserum by coated isotype-specific capture antibodies and the detection of insulin binding using europium-labeled human insulin (HI-Eu). This reagent was shown to have a detection limit of 10(-16) mol. Among various applications this technique permitted an isotype-specific determination of the affinity distribution of polyclonal antibodies from individual mice.
Vos
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Department of Cell Biology, Immunology and Genetics, Erasmus University, Rotterdam, The Netherlands.
Benner
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R
D000818:Animals; D000906:Antibodies / Q000032:analysis*; D000911:Antibodies, Monoclonal / Q000032:analysis; D002417:Cattle; D005063:Europium; D005455:Fluorescent Antibody Technique; D006801:Humans; D007132:Immunoglobulin Isotypes / Q000032:analysis; D066298:In Vitro Techniques; D007328:Insulin / Q000276:immunology*; D013552:Swine
D000906:Antibodies; D000911:Antibodies, Monoclonal; D007132:Immunoglobulin Isotypes; D007328:Insulin; D005063:Europium