Identification of a Strain of Hybridoma Cell Secreting the Monoclonal Antibody against Porcine IFN-γ

[Objective] The study aimed to obtain the strain of hybridoma cell that was stable to secrete the monoclonal antibody(mAb) against porcine IFN-γ.[Method] BALB/c mice were first immunized for 4 times with prokaryotic expression protein His-PoIFN-γ and then SP2/0 myeloma cells were fused with spleen cells of the immunized mice according to standard procedure.And then,the purified GST-PoIFN-γ was used as the detecting antigen and the positive hybridoma clones were screened by indirect ELISA and limited dilutions.Finally,a hybridoma which stably secreted the specific mAb against porcine IFN-γ was obtained and its specificity was characterized by indirect ELISA and Western blot.[Result] A hybridoma which stably secreted the specific mAb against porcine IFN-γ was obtained in this experiment and named as B3 strain.The detection by indirect ELISA and Western blot showed that the mAb secreted by B3 strain could only react with the fused proteins of His-PoIFN-γ and GST-PoIFN-γand it could not react with ChIFN-γ,aHis and GST expressed in E.coli.The ELISA titers of the mAb mice ascites secreted by B3 strain were 1∶160 000,mAb subtype belonged to IgG1 type and the light chain was κ chain.[Conclusion] This study provided the technique foundation for establishing the detection method of porcine IFN-γ and studying the immunization statue and mechanism of the organism.