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.