[通过沉淀和免疫吸附纯化人纤连蛋白]

[Purification of human fibronectin by precipitation and immunoadsorption].

作者信息

Allain J P, Lejars A, Pham C T, Gaillandre A, Lee H H

出版信息

Rev Fr Transfus Immunohematol. 1983 Apr;26(2):123-34. doi: 10.1016/s0338-4535(83)80087-7.

Abstract

Two types of human plasma fibronectin concentrate were obtained by simple, large scale, precipitation methods. Concentrate 1 contains 90% pure fibronectin in a mainly dimeric form. This easy, high yield method provides a functional fibronectin suitable for intravenous therapeutic purposes in deficient patients. Concentrate 2 contains 95-98% pure fibronectin with minimal amounts of fibronectin monomer and degradation products. It seems well adapted to serum free cultures of human cells. A third preparation of an apparently pure fibronectin has also been obtained by immuno-adsorption with a Sepharose 4B column to which has been coupled an anti-fibronectin monoclonal antibody. The capture of dimer, monomers and degradation products by this antibody and their release at pH 2.4 has been demonstrated by gel transfer on nitrocellulose sheet revealed by another monoclonal anti-fibronectin antibody labelled with peroxidase.

摘要

通过简单的大规模沉淀法获得了两种类型的人血浆纤连蛋白浓缩物。浓缩物1含有90%纯度的纤连蛋白，主要为二聚体形式。这种简便、高产率的方法提供了一种适用于缺乏患者静脉治疗用途的功能性纤连蛋白。浓缩物2含有95 - 98%纯度的纤连蛋白，纤连蛋白单体和降解产物含量极少。它似乎非常适合人细胞的无血清培养。通过用偶联了抗纤连蛋白单克隆抗体的琼脂糖4B柱进行免疫吸附，还获得了第三种明显纯的纤连蛋白制剂。用另一种用过氧化物酶标记的抗纤连蛋白单克隆抗体在硝酸纤维素膜上进行凝胶转移，已证明该抗体对二聚体、单体和降解产物的捕获及其在pH 2.4时的释放。