Prokaryotic Expression,Preparation and Characterization of Monoclonal Antibodies against Porcine IFN-α

The poIFN-α gene was cloned into a prokaryotic expression vector pET-28a,and a fusion expressed protein poIFN-α of 27 ku was obtained in E.coli.BALB/c mice were immunized intraperitoneally with purified poIFN-α protein.Murine myeloma cells SP2/0 were fused with the splenocytes of the immunized mice after the third immunization.An indirect ELISA was used to screen hybridomas for production of specific antibody in hybridoma cell.One hybridomas clone,named 2H12,which produced McAb steadily was obtained.The McAb showed strong reactivity with poIFN-α protein expressed by Western-blotting.The McAb belongs to IgG1 subtype,and its light chain was κ chain,which was subtyped by ELISA using acommercial kit.The McAb against by poIFN-α protein could be used for further analysis on the structure and function of poIFN-α.