[Preparation of 125I-labelled monoclonal antibodies of the insulin receptor]

C R Seances Soc Biol Fil. 1988;182(2):167-76.
[Article in French]

Abstract

Three monoclonal anti-insulin receptor antibodies have been labelled with 125I according to various methods (Cloramine T, Lactoperoxidase and IODO-GEN). The effect of labelling on antibody structure and function has been characterized using the following parameters: a) specific activity obtained in four different labelling procedures, at least; b) TCA labelled antibody precipitable 90 days after labelling; c) interaction between labelled antibodies and the insulin receptor; d) ability of antibodies to inhibit insulin-stimulated receptor auto-phosphorylation. Cloramine T method produced labelled antibody with constant specific activity; however, some preparations were unstable and showed reduced capacity to recognize the insulin receptor. Lactoperoxidase method produced stable antibodies; however, specific activity was highly variable and antibodies had low capacity to interact with the insulin receptor. The IODO-GEN method produced antibodies with constant specific activity, stable, high capacity to interact with the insulin receptor, and, moreover, maintaining in full the capacity to inhibit the insulin-stimulated auto-phosphorylation of the insulin receptor, since it does not induce antibody alterations which in turn affect antibody-receptor interaction biological action.

Publication types

  • English Abstract

MeSH terms

  • Antibodies, Monoclonal / immunology*
  • Antibodies, Monoclonal / metabolism
  • Insulin Antibodies / immunology*
  • Iodine Radioisotopes* / metabolism
  • Isotope Labeling / methods*
  • Phosphorylation
  • Protein Binding
  • Receptor, Insulin / immunology*
  • Receptor, Insulin / metabolism

Substances

  • Antibodies, Monoclonal
  • Insulin Antibodies
  • Iodine Radioisotopes
  • Receptor, Insulin