Yatohgo T, Izumi M, Kashiwagi H, Hayashi M
Institute of Clinical Medicine, University of Tsukuba, Ibaraki, Japan.
Cell Struct Funct. 1988 Aug;13(4):281-92. doi: 10.1247/csf.13.281.
The glycoprotein vitronectin (also called S-protein, serum spreading factor, or epibolin) promotes spreading of a variety of cultured cells, inhibits the cytotoxicity of membrane attack complex C5b-9, and modulates thrombin-antithrombin III activity. We developed a strikingly simple method to purify vitronectin from human plasma by heparin affinity chromatography. Serum was obtained from plasma by adding calcium and then centrifuging. The heparin-binding activity of vitronectin in human serum was activated with 8 M urea. The activated vitronectin specifically bound to heparin-Sepharose in 8 M urea and was eluted with 0.5 M NaCl containing 8 M urea. This procedure resulted in an approximately 250-fold purification of vitronectin with a 15-30% recovery; 3-6 mg of pure vitronectin were obtained from 100 ml human plasma within 2 days. The purified vitronectin preparations promoted spreading of BHK fibroblastic cells on substrates with a half-maximal activity at only 0.1 microgram/ml. This new method is very simple, rapid, inexpensive, and flexible. It could probably be readily scaled up for commercial applications.
糖蛋白玻连蛋白（也称为S蛋白、血清扩散因子或表皮调节素）可促进多种培养细胞的铺展，抑制膜攻击复合物C5b-9的细胞毒性，并调节凝血酶-抗凝血酶III活性。我们开发了一种极为简单的方法，通过肝素亲和色谱从人血浆中纯化玻连蛋白。通过添加钙然后离心从血浆中获得血清。人血清中玻连蛋白的肝素结合活性用8 M尿素激活。激活的玻连蛋白在8 M尿素中特异性结合到肝素-琼脂糖上，并用含8 M尿素的0.5 M NaCl洗脱。此过程使玻连蛋白得到了约250倍的纯化，回收率为15%-30%；在2天内从100 ml人血浆中获得了3-6 mg纯玻连蛋白。纯化的玻连蛋白制剂可促进BHK成纤维细胞在底物上的铺展，半最大活性仅为0.1微克/毫升。这种新方法非常简单、快速、廉价且灵活。它很可能易于扩大规模用于商业应用。
