Production of antibodies that inhibit the binding of insulin to its receptor |
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Authors: | Richard A Roth KY Wong Betty A Maddux ID Goldfine |
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Institution: | 1. Cell Biology Research Laboratory Nount Zion Hospital and Medical Center San Francisco, CA 94120 USA;2. Department of Medicine, Mount Zion Hospital and Medical Center San Francisco, CA 94120 USA;3. Department of Medicine University of California, San Francisco, USA;4. Department of Physiology University of California, San Francisco, USA |
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Abstract: | In this study, we report a procedure for producing antisera that block the binding of 125I-insulin to its receptor. After 2 injections with intact IM-9 cultured human lymphocytes, the antisera from 8 of 17 mice inhibited the binding of 125I-insulin to its receptor on IM-9 cells by 50% or greater. One antiserum at dilutions of 1:200 and 1:50 inhibited the binding of 125I-insulin by 50% and 80%, respectively. Four lines of evidence indicated that the inhibition of 125I-insulin binding by this antiserum was due to a specific immunoglobulin directed against the insulin receptor. First, removal of the immunoglobulin fraction of the antiserum resulted in a complete loss of its inhibitory activity. Second, the antiserum inhibited the binding of 125I-insulin to its receptor on both human cultured lymphocytes and human placenta particles. Third, the antisera bound solubilized insulin-receptor complexes. Finally, the antiserum did not inhibit the binding of 125I-human growth hormone to its receptor on IM-9 lymphocytes. These studies demonstrate therefore, a simple method for producing antibodies that block the binding of 125I-insulin to the human insulin receptor. |
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