Monoclonal Antibody to Human β Interferon: Characterization and Application

Abstract

Seven stable mouse hybridomas secreting monoclonal antibodies to human fibroblast β interferon (IFN-β) were isolated, all seven of which belonged to IgG1 subclass and κ type. While neutralizing the antiviral activity of human fibroblast IFN-β, they failed to neutralize both that of human IFN-α and human IFN-γ. These monoclonal antibodies neutralized the antiviral activity of human fibroblast IFN-β but not that of human IFN-α and IFN-γ. Two of the seven monoclonal antibodies, YSB-1 and YSB-2, showed particularly high neutralization titers in the ascitic fluid. Monoclonal antibodies were purified from cultures of hybridoma grown in a serum-free medium. The purified monoclonal antibodies, YSB-1(5μg/ml) and YSB-2(1μg/ml), neutralized IFN-β from 10EU/ml to 1EU/ml. Human fibroblast IFN-β was purified to 3.5xl0⁷IU/mg protein (1.Ox10⁸IU/mg protein in the peak fraction) by the monoclonal antibody (YSB-2) affinity column chromatography, whereas human recombinant IFN-β obtained from E.coli was also purified to 6.5x10⁷IU/mg protein (1.1x10⁸IU/mg protein in the peak fraction) by the monoclonal antibody (YSB-1) affinity column chromatography.