孕血清中早孕因子的分离纯化

Isolation and purifaction of early pregnancy factor (EPF) from human pregnancy sera

目的:通过离子交换层析和亲和层析的分离方法从孕血清中得到纯化的早孕因子(EPF),为进一步制备单克隆抗体做准备。方法:采用依次经过DEAE52阴离子交换层析,SP Sepharose F.F阳离子交换层析,Con ASepharose4B亲和层析和Heparin Sepharose Cl6B亲和层析分离纯化方案,最终得到具有早孕因子活性的Hepa-rin-Ⅱ成分,早孕因子活性采用活性玫瑰花环抑制实验检测,采用SDS-聚丙烯酰胺凝胶电泳鉴定纯化物。结果:SDS-聚丙烯酰胺凝胶电泳显示两条带,相对分子量分别为46.30kDa和10.71kDa。结论:经过四步层析得到较纯的EPF样品,并认为EPF在血清中可以以单体和多聚体形式存在。

Objective： To get the pure EPF from the pregnancy sera through ion exchange chromatography and affinity chromatography preparing for gaining the monoclonal antibody of EPF. Methods： The purification scheme involved sequential DEAE 52 chromatography, SP Sepharose F. F chromatography, Con A Sepharose 4B chromatography and Heprin Sepharose C1 6B chromatography. The biological activity was detected by rosette inhibition test. It was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS-PAGE）. Results： Two bands were observed on SDS-PAGE profile and the molecular weight were 46.30 KDa and 10.71 KDa. Conclusion： Get the pure EPF sample through four steps chromatography and think the EPF exists at oligomeric form and polymeric form in the pregnancy sera.