Preparation and characterization of a monoclonal antibody against porcine interferon gamma

BALB/c mice were immunized three times with purified recombinant porcine interferon-gamma expressed in E.coli. Hybridomas were produced by fusing spleen cells from the immunized BALB/c mouse with SP2/0 myeloma cells according to standard procedure. Positive clones were screened by indirect ELISA and limited dilutions. Finally, a hybridoma which stably secrets specific monoclonal antibody against porcine IFN-|? was established. The monoclonal antibody, named B11 strain, was shown to strongly react with commercial porcine IFN-|? standard in Western blot and indirect ELISA. The B11 strain also showed strong reactivity in immunofluorescent antibody test on the porcine PBMCs stimulated with PMA. Further characterization revealed that B11 strain belongs to IgG1 isotype, and the light chains are |e chain.