人,牛内皮细胞组织型纤溶酶原激活物活性的比较

体外培养人脐静脉及胎牛主动脉内皮细胞,分别从其条件培养液中提取t-PA。实验主要分三步:1)胎牛内皮细胞t-PA的纯化;2)兔抗牛内皮细胞t-PA抗血清的制备及纯化其1gG组分;3)免疫亲和层析、纯化人脐静脉及胎牛主动脉内皮细胞t-PA。结果表明,用微量法免疫动物制备的兔抗牛内皮细胞t-PA抗血清具有较高的滴度与特异性。将该抗血清1gG与人、牛内皮细胞CM交联免疫亲和层析,可获得电泳纯的t-PA,其分子量为65000及67000道尔顿。两者活性比较,胎牛主动脉内皮细胞t-PA活性比人脐静脉内皮细胞t-PA明显增高。

Comparison of t-PA activity of endothelial cells from human and bovine

Endothelial cells from human umbilical vein and fetal bovine aorta werecultured in vitro. Their tissue plasminogen activator (t-PA) were purified from the condi-tioned media (CM) collected separately. There were three steps in the experiment. 1.Purification of t-PA from fetal bovine aorta endothelicel cells (FBAEC). 2.Preparation of rabbit anti-FBAEC t-PA antiserum and purification of its IgGfraction. 3.Further purification of t-PA secreted by human umbilical vein endothelia cells(HUVEC) and FBAEC by immunoaffinity chromatography. The results showed that rab-bit anti-bovine ECs t-PA antiserum prepared by microinjection method possess bothhigher titre and specificity. By using the IgG fraction crosss linked on sepharose 4B anelectrophoretic purified purity t-PA can be obtained from human or bovine ECs CM. Themoleculor weight of t-PA from HUVEC and FBAEC was 65KD or 67KD respectively.The activity of t-PA from FBAEC was much higher than that from HUVEC.