PLATE BINDING ASSAY FOR MONOCLONAL ANTI–RECEPTOR ANTIBODIES

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Abstract
A new procedure has been developed for rapid identification of monoclonal antibodies to the insulin receptor. The technique uses the ability of polyvinyl chloride plates coated with anti-mouse antibodies to specifically adsorb antibodies in hybridoma supernatants. Detection of anti-receptor antibodies is accomplished by allowing the adsorbed anti-receptor antibodies to bind radiolabeled hormone-receptor complexes. The method does not require pure receptor and can be adapted for studies of any receptor or binding protein where a labeled ligand is available. With small amounts of the insulin receptor (20-50 ng), the method could detect antibody concentrations as low as 300 pM. In summary, the assay is sensitive, rapid, and requires small amounts of impure receptor.

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