On-column refolding of corticotropin-releasing factor receptor 1 extracellular domain by size exclusion chromatography

被引：5

作者：

Ersoy, Maria ^{[1
]}

Matic, Sandra ^{[1
]}

Schmidt, Stefan R. ^{[1
]}

机构：

[1] AstraZeneca R&D Sodertalje, Global Prot Sci & Supply, SE-15185 Sodertalje, Sweden

来源：

BIOMEDICAL CHROMATOGRAPHY | 2009年 / 23卷 / 10期

关键词：

refolding; chromatography; on-column; corticotropin-releasing factor receptor 1; INCLUSION-BODY PROTEINS; ESCHERICHIA-COLI; CRF RECEPTOR; PURIFICATION; EXPRESSION;

D O I：

10.1002/bmc.1233

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Expressing the extracellular domain of corticotropin releasing factor receptor 1 in Escherichia coli usually results in the formation of inclusion bodies. Here we describe the optimization of refolding by applying size exclusion chromatography with a denaturing guanidine hydrochloride gradient and a refolding buffer containing glycerol. Several chromatographic parameters like gradient length, flow rate, sample concentration and chromatography resin characteristics were evaluated. Recovery yields of refolded protein above 50% using a Superdex 200 column demonstrate the usefulness of this method. Copyright (C) 2009 John Wiley & Sons, Ltd.

引用

页码：1121 / 1128

页数：8

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