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[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.

DOI:10.1016/s0338-4535(83)80087-7
PMID:6348922
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。

相似文献

1
[Purification of human fibronectin by precipitation and immunoadsorption].[通过沉淀和免疫吸附纯化人纤连蛋白]
Rev Fr Transfus Immunohematol. 1983 Apr;26(2):123-34. doi: 10.1016/s0338-4535(83)80087-7.
2
The preparation of plasma fibronectin antigen and antiserum.血浆纤连蛋白抗原及抗血清的制备。
J Immunol Methods. 1981;43(1):29-33. doi: 10.1016/0022-1759(81)90033-8.
3
[Human plasma fibronectin. Comparison of methods for preparation of a concentrate for therapeutic use from different sources].[人血浆纤连蛋白。从不同来源制备治疗用浓缩物的方法比较]
Rev Fr Transfus Hemobiol. 1990 Dec;33(6):391-405. doi: 10.1016/s1140-4639(05)80056-5.
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[The use of immobilized antibodies against fibronectin for isolation of macromolecular complexes of fibronectin with other secreted proteins from human fibroblasts].[使用抗纤连蛋白的固定化抗体从人成纤维细胞中分离纤连蛋白与其他分泌蛋白的大分子复合物]
Vopr Med Khim. 1988 Jul-Aug;34(4):101-4.
5
A simplified procedure for the preparation of antibodies to serum fibronectin.一种制备抗血清纤连蛋白抗体的简化程序。
J Immunol Methods. 1980;34(2):155-65. doi: 10.1016/0022-1759(80)90169-6.
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Affinity purification of human plasma fibronectin on immobilized gelatin.在固定化明胶上亲和纯化人血浆纤连蛋白。
J Chromatogr. 1988 Nov 18;432:93-102. doi: 10.1016/s0378-4347(00)80636-2.
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Phagocytosis-promoting activity of avian plasma and fibroblastic cell surface fibronectins.禽血浆和成纤维细胞表面纤连蛋白的促吞噬活性。
Mol Cell Biochem. 1981 May 26;36(3):147-55. doi: 10.1007/BF02357031.
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Purification of Human Plasma/Cellular Fibronectin and Fibronectin Fragments.人血浆/细胞纤连蛋白及纤连蛋白片段的纯化
Methods Mol Biol. 2017;1627:309-324. doi: 10.1007/978-1-4939-7113-8_20.
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Plasma fibronectin: three steps to purification and stability.血浆纤连蛋白:纯化与稳定的三个步骤。
Protein Expr Purif. 1999 Oct;17(1):146-52. doi: 10.1006/prep.1999.1103.
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Immunochemical analysis of fibronectin using monoclonal antibodies.使用单克隆抗体对纤连蛋白进行免疫化学分析。
Biochim Biophys Acta. 1982 Aug 10;705(3):330-40. doi: 10.1016/0167-4838(82)90255-2.