一种用于高效亲和色谱的新型基质及其在抗凝血酶III纯化中的应用。

A novel matrix for high performance affinity chromatography and its application in the purification of antithrombin III.

作者信息

Zhao Rui, Luo Jia, Shangguan Dihua, Liu Guoquan

机构信息

Center for Molecular Science, Institute of Chemistry, Chinese Academy of Sciences, Zhongguancun, 100080 Beijing, P.R China.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2005 Feb 25;816(1-2):175-81. doi: 10.1016/j.jchromb.2004.11.030.

DOI:10.1016/j.jchromb.2004.11.030

PMID:15664348

Abstract

Viscose fiber, a regenerated cellulose, was evaluated for using as a novel matrix for high performance affinity chromatography. With a one-step activation with epichlorohydrin, heparin can be readily covalently attached to the matrix. This heparin-viscose fiber material was used for purifying antithrombin III (AT III) from human plasma. The purity of the AT III from this one-step purification is 93% as measured by SDS-PAGE and the protein recovery yield is about 90%. This column is highly specific as described by the dissociation constant of the complex of immobilized heparin and AT III, which was 2.83 x 10(-5)mol/L. And more important, this viscose fiber material demonstrated its excellent mechanical property that allows the flow rate to reach up to 900 cm/h or more.

摘要

粘胶纤维，一种再生纤维素，被评估用作高性能亲和色谱的新型基质。通过用环氧氯丙烷一步活化，肝素可以很容易地共价连接到基质上。这种肝素-粘胶纤维材料用于从人血浆中纯化抗凝血酶III（AT III）。通过SDS-PAGE测定，该一步纯化得到的AT III纯度为93%，蛋白质回收率约为90%。如固定化肝素与AT III复合物的解离常数所示，该柱具有高度特异性，为2.83×10(-5)mol/L。更重要的是，这种粘胶纤维材料表现出优异的机械性能，允许流速达到900 cm/h或更高。