Purification and on-column refolding of EGFP overexpressed as inclusion bodies in Escherichia coli with expanded bed anion exchange chromatography

被引：29

作者：

Cabanne, C ^{[1
]}

Noubhani, AM ^{[1
]}

Hocquellet, A ^{[1
]}

Dole, R ^{[1
]}

Dieryck, W ^{[1
]}

Santarelli, X ^{[1
]}

机构：

[1] Univ Bordeaux 2, Ecole Super Technol Biomol Bordeauc, F-33076 Bordeaux, France

来源：

JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2005年 / 818卷 / 01期

关键词：

expanded bed adsorption; EGFP; purification; chromatography; refolding; inclusion bodies;

D O I：

10.1016/j.jchromb.2004.10.023

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The enhanced green fluorescent protein (EGFP) was over-expressed in Escherichia coli as inclusion bodies to increase its quantity and to facilitate its purification. Insoluble EGFP has been purified on Q Hyper Z matrix by expanded bed adsorption after solubilization in 8 M urea. The adsorption was made in expanded bed mode to avoid centrifugation. EBA-column refolding was done by elimination of urea and elution with NaCl. The EGFP was obtained as a highly purified soluble form with similar behavior in fluorescence and electrophoresis as native EGFP. (C) 2004 Elsevier B.V. All rights reserved.

引用

页码：23 / 27

页数：5

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