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[冻干明胶-琼脂糖组合瓣膜在从人血浆中选择性提取纤连蛋白中的应用]

[Valve of a freeze-dried gelatin-agarose combination in selective extraction of fibronectin from human plasma].

作者信息

Burnouf T, Allary M, Guilbaud J, Boschetti E, Saint-Blancard J

出版信息

Rev Fr Transfus Immunohematol. 1981 Dec;24(6):559-70. doi: 10.1016/s0338-4535(81)80052-9.

Abstract

We report here a simple procedure for the isolation of human plasma fibronectin by affinity adsorption on a new lyophilized adsorbent which is obtained by mixing and polymerizing agarose with gelatin. This method permits to obtain fibronectin and a fibronectin-depleted plasma without dilution. The yield of fibronectin is about 20 to 25 per cent. Fibronectin was identified by cross-reactions against specific antibodies. Its purity (greater than or equal to 99 per cent) was controlled by immunoelectrophoresis and SDS-PAGE. Furthermore, its biological activity was demonstrated by a spreading test performed on BHK cells. Such a test can be used to determine the specific activity of extracted fibronectin considering that the number of spread cells placed in presence of exogenous fibronectin (5 microgram/ml of culture medium) was 4.5 to 5 fold-superior to the control, after 30 minutes of incubation at 37 degrees C.

摘要

我们在此报告一种通过亲和吸附于一种新型冻干吸附剂来分离人血浆纤连蛋白的简单方法,该吸附剂是通过将琼脂糖与明胶混合并聚合而获得的。此方法能够在不稀释的情况下获得纤连蛋白和去除纤连蛋白的血浆。纤连蛋白的产率约为20%至25%。通过与特异性抗体的交叉反应鉴定了纤连蛋白。其纯度(大于或等于99%)通过免疫电泳和SDS-PAGE进行控制。此外,通过在BHK细胞上进行的铺展试验证明了其生物活性。考虑到在37℃孵育30分钟后,在外源纤连蛋白(5微克/毫升培养基)存在下铺展的细胞数量比对照高4.5至5倍,这样的试验可用于确定提取的纤连蛋白的比活性。

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