孕牛血清中早孕因子的分离纯化

PURIFICATION OF EARLY PREGNANCY FACTOR FROM BOVINE SERA

采用DEAE Sepharose(DEAE 琼脂糖凝胶 )、CM Sepharose(CM 琼脂糖凝胶 )等离子交换层析、抗孕牛全血清IgG Sepharose 4B亲和层析等技术 ,分离纯化妊娠 4～ 5个月健康孕牛全血清中的早孕因子 (EPF) ,用玫瑰花环抑制实验对各阶段产物EPF活性进行检测。结果显示 ,经层析过柱后的CM ⅡA提取物有EPF活性。经SDS PAGE银染显色证实它含有分子量为 2 1、6 7、80KD的蛋白 ,实验结果支持 2 1KD的多肽为主要EPF活性物成分。同时实验表明了亲和层析对进一步纯化EPF是可行的。

To purify early pregnancy factor(EPF) from the pregnant bovine serum of 45 months,we used a series of techniques,such as DEAESepharose,CMSepharose ionexchange chromatography,etc.EPF activity of each fraction was determined by the rosette inhibition test(RIT).The result showed that the fraction CMIIA has EPF activity.When the pattern of this SDSPAGE was compared with the bands obtained from nonpregnancy serum,these bands were not found,we thought that the bovine EPF active material of M.W.21KD might be the major active form of the bovine EPF in this gestation period.In adddition,the affinity gel (Sepharose4B) which was coupled with rabbit anti pregnancy bovine whole serum IgG was proved effective for the purification of EPF.