Purification and further characterization of an anti-murine interferon-gamma monoclonal neutralizing antibody.
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.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification*; D001202:Ascitic Fluid / Q000032:analysis; D002848:Chromatography, DEAE-Cellulose; D002851:Chromatography, High Pressure Liquid; D006825:Hybridomas / Q000378:metabolism; D007371:Interferon-gamma / Q000276:immunology*; D051379:Mice; D009500:Neutralization Tests; D051381:Rats; D014779:Virus Replication / Q000187:drug effects
D000911:Antibodies, Monoclonal; D007371:Interferon-gamma