DEVELOPMENT OF MONOCLONAL ANTIBODIES AGAINST RECOMBINANT BOVINE INTERFERON-GAMMA

Seven week-old BALB/c mice were immunized with purified fusion protein rHis-BoIFN-γ that was obtained from a prokaryotic expression system.Splenocytes from immunized mice were fused with Sp2/0-Ag-14 myeloma cells.The supernatants of hybridoma clones were screened by indirect ELISA using purified rGST-BoIFN-γ.Thirteen hybridoma cell lines secreting MAbs against BoIFN-γ were obtained and assigned as 1C12,1F7,1G5,3E6,4D5,5E11,5G4,6F8,6G6,7E9,8D3,8F8 and 9G11.The immunoglobulin subclass of MAb 5G4 was IgG2b and all others were IgG1.The ascitic titers of these MAbs were 640 000,160 000,640 000,320 000,160 000,320 000,2 560 000,2 560 000,320 000,640 000,80 000,2 560 000 and 10 000,respectively.In Dot-ELISA,all MAbs specifically reacted with the immunogen and the screening antigen.Western blot analysis confirmed that all MAbs reacted with the corresponding recombinant proteins.The MAbs also reacted with the standard recombinant bovine IFN-γ with biological activity.The availability of these MAbs provides a basis for further research and clinical applications of BoIFN-γ in pathogenesis and diagnosis of bovine diseases.