Preparation Of The Monoclonal Antibodies Against Human Early Pregnancy Factor

Object:The recombinant protein of GST-EPF11 and BSA-MC29 immunized Balb/c mice for preparation and identification of the monoclonal antibodies.The monoclonal antibodies were obtained. Methods:The early pregnancy factor lacking the immunodepression site was amplified using the pMD18T-HSPE1 plasmid as template by PCR. The product digested by BamHI and Xhol was ligased with a prokaryotic expression vector pGEX-6P-1 to construct the recombinant expression plasmid pGEX6P-EPF11. Escherichia coli BL21 transformed with pGEX6P-EPF11 was induced by 1 mmol/L IPTG to express the intersted protein.At the same time we designed a polypeptide, which is coupled with carrier protein bovine serum abumin (BSA) to prepare immunizing antigen BSA-MC29. Balb/c mice was immunized by the purified recombinant protein and BSA-MC29 for the preparation of the monoclonal antibodies, then the monoclonal antibodies was identified by using ELISA. Results:The recombinant expression plasmid pGEX6P-EPF11 was successfully constructed, and the recombinant protein GST-EPF11 was expressed. The results of SDS-PAGE and Western-blotting demonstrated that the recombinant protein of GST-EPF11 with a molecular weight of 35 kDa was expressed successfully, which existed partially as soluble protein.The immunized Balb/c mice was fused. We obtained the monoclonal antibodies of the recombinant protein and BSA-MC29 from immunized Balb/c mice. Conclusion:The recombinant protein GST-EPF11 and MC29-BSA was obtained, the immunized Balb/c mice and antibody effectiveness was analyzed. The monoclonal antibodies of BSA-MC29 and the recombinant protein that expressed by Early Pregnancy Factor were obtained.