Refolding of recombinant human interferon alpha-2a from Escherichia coli by urea gradient size exclusion chromatography

doi:10.1134/S0003683813010055

	Refolding of recombinant human interferon alpha-2a from Escherichia coli by urea gradient size exclusion chromatography
	Gao, F.1; Shi, L.1,2; Xu, L. -X.1
刊名	APPLIED BIOCHEMISTRY AND MICROBIOLOGY
	2013-01
卷号	49 期号:1 页码:11-17
ISSN号	0003-6838
DOI	10.1134/S0003683813010055
文献子类	Article
英文摘要	Protein refolding is still a puzzle in the production of recombinant proteins expressed as inclusion bodies (IBs) in Escherichia coli. Gradient size exclusion chromatography (SEC) is a recently developed method for refolding of recombinant proteins in IBs. In this study, we used a decreasing urea gradient SEC for the refolding of recombinant human interferon alpha-2a (rhIFN alpha-2a) which was overexpressed as IBs in E. coli. In chromatographic process, the denatured rhIFN alpha-2a would pass along the 8.0-3.0 M urea gradient and refold gradually. Several operating conditions, such as final concentration of urea along the column, gradient length, the ratio of reduced to oxidized glutathione and flow rate were investigated, respectively. Under the optimum conditions, 1.2 x 10(8) IU/mg of specific activity and 82% mass recovery were obtained from the loaded 10 ml of 1.75 mg/ml denatured protein, and rhIFN alpha-2a was also purified during this process with the purity of higher than 92%. Compared with dilution method, urea gradient SEC was more efficient for the rhIFN alpha-2a refolding in terms of specific activity and mass recovery.
资助项目	National Natural Science Foundation of China[30970640] ; S&T Plan Projects of Shandong Provincial Education Department[J06I04] ; Research Fund of Qufu Normal University[XJ200822/XJ201208] ; Knowledge Innovation Program of the Chinese Academy of Sciences[SIMM0912QN-10]
WOS关键词	MOVING-BED CHROMATOGRAPHY ; GEL-FILTRATION ; PROTEIN ; LYSOZYME ; PURIFICATION ; RENATURATION ; EXPRESSION ; SYSTEM ; YIELD
WOS研究方向	Biotechnology & Applied Microbiology ; Microbiology
语种	英语
出版者	PLEIADES PUBLISHING INC
WOS记录号	WOS:000312492300002
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/277830]
专题	中国科学院上海药物研究所
通讯作者	Gao, F.
作者单位	1.Qufu Normal Univ, Coll Life Sci, Qufu 273165, Peoples R China; 2.Chinese Acad Sci, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China
推荐引用方式 GB/T 7714	Gao, F.,Shi, L.,Xu, L. -X.. Refolding of recombinant human interferon alpha-2a from Escherichia coli by urea gradient size exclusion chromatography[J]. APPLIED BIOCHEMISTRY AND MICROBIOLOGY,2013,49(1):11-17.
APA	Gao, F.,Shi, L.,&Xu, L. -X..(2013).Refolding of recombinant human interferon alpha-2a from Escherichia coli by urea gradient size exclusion chromatography.APPLIED BIOCHEMISTRY AND MICROBIOLOGY,49(1),11-17.
MLA	Gao, F.,et al."Refolding of recombinant human interferon alpha-2a from Escherichia coli by urea gradient size exclusion chromatography".APPLIED BIOCHEMISTRY AND MICROBIOLOGY 49.1(2013):11-17.