Purification and further characterization of an anti-murine interferon-gamma monoclonal neutralizing antibody

J Interferon Res. 1986 Oct;6(5):489-97. doi: 10.1089/jir.1986.6.489.

Abstract

The R4-6A2 rat-mouse hybridoma, which secretes a rat IgG1 monoclonal antibody (MAb) capable of neutralizing murine interferon-gamma (MuIFN-gamma), was grown as ascites in athymic nude (nu/nu) mice. The neutralizing titers of ascitic fluids were 400 times higher than those of supernatants from R4-6A2 cultures. Moreover, the specific activities (MAb) neutralizing units/mg protein) of the ascitic fluids were much greater than those of R4-6A2 culture supernatants. Ascitic fluid MAb was purified by anion-exchange chromatography, with excellent recovery of applied neutralizing activity. The ability of the R4-6A2 anti-MuIFN-gamma MAb to neutralize several different activities of MuIFN-gamma was studied. The MAb neutralized the ability of MuIFN-gamma to inhibit the growth of cells (antiproliferative activity). The ability of MuIFN-gamma to render cells of a heterologous species (rat) resistant to viral replication was also neutralized by this MAb. The MAb-neutralizing titers for MuIFN-gamma antiviral activities, on both homologous (murine L929) and heterologous (rat fibroblast) cells, were inversely proportional to the antiviral activity titers on each cell type.

Publication types

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

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology
  • Antibodies, Monoclonal / isolation & purification*
  • Ascitic Fluid / analysis
  • Chromatography, DEAE-Cellulose
  • Chromatography, High Pressure Liquid
  • Hybridomas / metabolism
  • Interferon-gamma / immunology*
  • Mice
  • Neutralization Tests
  • Rats
  • Virus Replication / drug effects

Substances

  • Antibodies, Monoclonal
  • Interferon-gamma