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

被引：27
|
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
收藏
页码：23 / 27
页数：5
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