Large-scale, one-step purification of murine interferon-beta using a monoclonal antibody.
A monoclonal antibody against murine interferon-beta (MuIFN-beta) was prepared using standard methods. Antibodies were immobilized by coupling to Sepharose and used for large-scale purification of poly(I) . poly(C)-induced mouse L cell IFN. Antibodies isolated from the serum of one nude mouse which was transplanted with the anti-MuIFN-beta antibodies producing hybridoma were able to bind at least 7 X 10(7)U MuIFN-beta. In one single antibody affinity chromatography step MuIFN-alpha was separated from MuIFN-beta and a 1000-fold purification of MuIFN-beta was obtained. The purified material had a specific activity of 5 X 10(8)U/mg protein. The recovery from the antibody column was 100%. SDS-PAGE analysis of the purified material revealed the presence of one single protein band with a molecular weight of 33 kD, representing MuIFN-beta.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D066298:In Vitro Techniques; D007370:Interferon Type I / Q000276:immunology / Q000302:isolation & purification*; D007739:L Cells / Q000276:immunology; D051379:Mice
D000911:Antibodies, Monoclonal; D007370:Interferon Type I