Preparation of the specific monoclonal antibody against bovine γ-interferon and its properties

Two hybridoma cells secreting monoclonal antibodies against bovine γ-interferon were obtained by fusion between SP2/0 cells and spleen cells from mice immunized with GST-BovIFN-γ fusion protein expressed on IPTG-induced E.coli.This fusion protein was recovered from gel of SDS-PAGE,and used as immunogen to immunize BALB/c mice.The monoclonal antibodies thus obtained were designated as BovIFN-γ4A3 and BovIFN-γ4G5 respectively.As demonstrated by ELISA and IFA,these monoclonal antibodies reacted only with BovIFN-γ,but not with GST or chicken IFN-γ.In addition,they could neutralize the cytopathic effect on bovine kedney cells induced by vesicular stomatitis virus(VSV) and showed high specificity and stability as demonstrated in indirect ELISA,Western blotting and IFA assays.It is evident that these monoclonal antibodies can be used for the surveillance and control of bovine tuberculosis in milk cows.