A mild hydrophobic interaction chromatography involving polyethylene glycol immobilized to agarose media refolding recombinant Staphylococcus aureus elongation factor G

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	A mild hydrophobic interaction chromatography involving polyethylene glycol immobilized to agarose media refolding recombinant Staphylococcus aureus elongation factor G
	Li, JJ ; Venkataramana, M ; Wang, AQ ; Sanyal, S ; Janson, JC ; Su, ZG
刊名	PROTEIN EXPRESSION AND PURIFICATION
	2005-04-01
卷号	40 期号:2 页码:327-335
关键词	elongation factor G refolding immobilized polyethylene glycol hydrophobic interaction chromatography pulse elution urea
ISSN号	1046-5928
英文摘要	Recombinant Staphylococcus aureus elongation factor G (EF-G) is difficult to refold by dilution due to the formation of large amounts of misfolded structures. However, refolding of EF-G by adsorption to a chromatographic column packed with immobilized polyethylene glycol 20,000 (PEG 20 K) followed by pulse elution with 8 M urea resulted in 88% mass recovery and 80% of correctly refolded structure. The PEG 20 K was coupled to brominated allyl group derivatized Sepharose High Performance to construct a mild hydrophobic adsorbent. Various other hydrophobic interaction adsorbents were also attempted to retold EF-G. However, ligands with high hydrophobicity tended to misfold EF-G, resulting in irreversible adsorption. Various solvents, detergents, and low temperature as well as 8 M urea were tried to release bound EF-G. Only pulse elution with 8 M urea was efficient. Urea concentrations favorable for efficiently refolding EF-G were investigated. Low urea concentration produced more misfolded structures. (c) 2005 Elsevier Inc. All rights reserved.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
研究领域[WOS]	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
关键词[WOS]	INCLUSION-BODIES ; FUSIDIC ACID ; LYSOZYME ; AGGREGATION ; PROTEINS
收录类别	SCI
语种	英语
WOS记录号	WOS:000227889600013
公开日期	2013-09-20
内容类型	期刊论文
版本	出版稿
源URL	[http://ir.ipe.ac.cn/handle/122111/1582]
专题	过程工程研究所_生化工程国家重点实验室
作者单位	1.Chinese Acad Sci, Natl Key Lab Biochem Engn, Inst Proc Engn, Beijing 100080, Peoples R China 2.Uppsala Univ, Dept Cell & Mol Biol, BMC, SE-75124 Uppsala, Sweden 3.Uppsala Univ, Dept Surface Biotechnol, BMC, SE-75123 Uppsala, Sweden
推荐引用方式 GB/T 7714	Li, JJ,Venkataramana, M,Wang, AQ,et al. A mild hydrophobic interaction chromatography involving polyethylene glycol immobilized to agarose media refolding recombinant Staphylococcus aureus elongation factor G[J]. PROTEIN EXPRESSION AND PURIFICATION,2005,40(2):327-335.
APA	Li, JJ,Venkataramana, M,Wang, AQ,Sanyal, S,Janson, JC,&Su, ZG.(2005).A mild hydrophobic interaction chromatography involving polyethylene glycol immobilized to agarose media refolding recombinant Staphylococcus aureus elongation factor G.PROTEIN EXPRESSION AND PURIFICATION,40(2),327-335.
MLA	Li, JJ,et al."A mild hydrophobic interaction chromatography involving polyethylene glycol immobilized to agarose media refolding recombinant Staphylococcus aureus elongation factor G".PROTEIN EXPRESSION AND PURIFICATION 40.2(2005):327-335.