Development Of Monoclonal Antibody Specific To Bovine Interferon-Gamma

Two monoclonal antibodies (McAbs) against BovIFN-gamma were developed by fusions between hybridoma SP2/0 cells and spleen cells from the mice immunized with GST-BovIFN-gamma fusion protein, which was expressed in recombinant E.coli induced by IPTG. In this study . interest protein was cut from the gel of SDS-PAGE, and the protein was used for immunizing the Balb/c mice. Specific McAbs were screened by indirect ELISA Two of McAbs were named as BovIFM -y-4A3 and BovIFN-y-4G5 respectively. The subtype of immunoglobulin is IgG2b for BovIFN -y-4A3 and IgM for BovlFN-y-4G5 in capture ELISA.. The ascites titer of the McAb BovIFN-y-4A3 was 1:1.3 107 and the McAb BovIFN-y-4G5 was 1:25600 in indirect ELISA. Western-blot analysis results showed that two McAbs could react with the GST-BovIFN-y fusion protein which was expressed in recombinant E.coli induced by IPTG, and not react with GST; Immunofluorescence Assay (IFA) results showed that the McAb BovlFN-y-4A3 could react with Sf9 cells expressing rBov-IFN-y. Neutralization test results indicated that the McAb had neutralizing-activities, and the neutralizing titer was above 1:6000.A sandwich enzyme-linked immuosorbant assay for rBovIFN-gamma was developed. BovIFN-y-4A3 was labeled with HRP by NaIO4 oxidation. BovIFN-y-4G5 was used as capture antibody. This method could check the rBovINF-y out and it will bevery useful for some diseases diagnosis.