特异性血管生成抑制素的制备及其抑制血管生成作用

目的：从血浆中纯化制备有特异抑制血管生成活性的天然ａｎｇｉｏｓｔａｔｉｎ蛋白，为研究和应用要下基础，方法：制备Ｌ－ｌｙｓｉｎｅ偶联的的Ｓｅｐｈａｒｏｓｅ４Ｂ亲和层析柱，从血浆中纯化纤溶酶原用弹性蛋白酶在体外进行有限消化，再经Ｌ－ｌｙｓｉｎｅＳｅｐｈａｒｏｓｅ４Ｂ亲和层析纯化获得天然ａｎｇｉｏｓｔａｔｉｎ片段，通过体外血管内皮细胞生长抑制分析和体内鸡胚尿囊膜血管生成抑制实验测定纯化蛋白活性。结果：以

Preparation of native angiostatin from human plasma and its antiangiogenesis effects

AIM: To develop a method to prepare native angiogenesis inhibitor angiostatin from human plasma for research and practical use. METHODS: L lysine linked Sepharose 4B was prepared by ligation of L lysine to CNBractivated Sepharose 4B. Affinity chromatograph with L lysine Sepharose 4B was utilized to purify plasminogen from human plasma and limited proteolysis by Elastase was applied to purified plasminogen. Angiostatin fragments were purified by using L lysine Sepharose 4B. BAE cell growth inhibition assay and CAM angiogenesis assay were carried out to check the activity of the purified angiostatin. RESULTS: Plasminogen was purified though affinity chromatograph with L lysine Sepharose 4B. Angiostatin was generated though limited proteolysis by Elastase and purified by affinity chromatograph with L lysine Sepharose 4B. Angiostatin had potent inhibitory effects on bovine aortic endothelial cell growth and CAM angiogenesis. CONCLUSION: Active native angiostatin is purified by our method.