Isolation of anti-idiotypic antibodies to T cells using an anti-framework determinant antibody

J Immunol Methods. 1987 Apr 16;98(2):219-26. doi: 10.1016/0022-1759(87)90008-1.

Abstract

A panel of anti-idiotypic antibodies to the T cell line HPB-ALL was produced by screening with a novel enzyme-linked immunoadsorption assay (ELISA). Using the beta framework I (beta F1) monoclonal antibody directed at a common determinant on the human T cell receptor beta subunit, we were able to specifically capture the receptor molecule from a cell lysate preparation and use this as the basis of an ELISA assay. Hybridoma supernatants were tested for their ability to bind to the receptor thus captured. A total of four antibodies were isolated by this method, and they were shown to immunoprecipitate a disulfide-linked heterodimer composed of alpha (49 kDa) and beta (40 kDa) subunits from HPB-ALL cells, similar to the subunits recognized by the beta F1 antibody. Furthermore, all four antibodies blocked the binding of T40/25, an anti-idiotype to HPB-ALL. Three of these antibodies blocked the binding of anti-Leu 4 to a similar degree as did T40/25, while one did not. This suggests that these new anti-idiotypic antibodies recognize distinct but associated idiotypic determinants. The isolation of such antibodies for any particular T cell line or tumor promises to be useful for biological studies of T cell malignancy in humans.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antibodies, Anti-Idiotypic / biosynthesis
  • Antibodies, Anti-Idiotypic / immunology
  • Antibodies, Anti-Idiotypic / isolation & purification*
  • Antibodies, Monoclonal
  • Binding, Competitive
  • Cell Line
  • Enzyme-Linked Immunosorbent Assay
  • Epitopes / analysis
  • Epitopes / immunology
  • Hybridomas / metabolism
  • Immunoglobulin Idiotypes / biosynthesis
  • Immunoglobulin Idiotypes / immunology
  • Immunoglobulin Idiotypes / isolation & purification*
  • Mice
  • Mice, Inbred BALB C
  • Precipitin Tests
  • Receptors, Immunologic / physiology
  • T-Lymphocytes / immunology*
  • T-Lymphocytes / ultrastructure

Substances

  • Antibodies, Anti-Idiotypic
  • Antibodies, Monoclonal
  • Epitopes
  • Immunoglobulin Idiotypes
  • Receptors, Immunologic