Immobilised metal-ion affinity chromatography purification of histidine-tagged recombinant proteins: a wash step with a low concentration of EDTA

被引：25

作者：

Westra, DF

Welling, GW

Koedijk, DGAM

Scheffer, AJ

The, TH

Welling-Wester, S

机构：

[1] Univ Groningen, Dept Med Microbiol, NL-9713 GZ Groningen, Netherlands

[2] Univ Groningen, Dept Clin Immunol, NL-9713 GZ Groningen, Netherlands

来源：

JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2001年 / 760卷 / 01期

关键词：

recombinant proteins; EDTA; histidine;

D O I：

10.1016/S0378-4347(01)00261-4

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Immobilised metal-ion affinity chromatography (IMAC) is widely used for the purification of recombinant proteins in which a poly-histidine tag is introduced. However, other proteins may also bind to IMAC columns. We describe the use of a washing buffer with a low concentration of EDTA (0.5 mM) for the removal of proteins without histidine tag from IMAC columns. Four histidine-tagged recombinant proteins/protein complexes were purified to homogeneity from cell culture medium of insect cells by using an EDTA washing buffer. The presence of a low concentration of EDTA in washing buffers during IMAC may have a general application in the purification of histidine-tagged proteins. (C) 2001 Elsevier Science B.V. All rights reserved.

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页码：129 / 136

页数：8

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