Preparation and Characterization of Monoclonal Antibody Against Bovine Interferon-γ

Recombinant fusion bovine IFN-γ expressed on IPTG-induced E.coli was purified by Glutathione Sepharose 4 Fast Flow affinity column.Balb/c mice were immunized 5 times with purified reccombinant BovIFN-γ protein.Murine myeloma cells were fused with splenocytes of the immunized mice.Positive clones were screened by indirect ELISA and limited dilutions.The main characteristics of screened McAb against BovIFN-γ were further identified by ELISA.Finally,a hybridoma stably secreting specific monoclonal antibody against BovIFN-γ was named A12E9.And the titers of cell culture supernatant and ascitic fluid were 1∶3 200,1∶160 000,respectively.