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Immobilized Triton X-100-assisted refolding of Green Fluorescent Protein-Tobacco Etch Virus protease fusion protein using beta-cyclodextrin as the eluent
Li, Jing Jing1; Wang, Ai-Qing1; Janson, Jan-Christer2; Ballagi, Andras2; Chen, Jing1; Liu, Yong-Dong1; Ma, Guang-Hui1; Su, Zhi-Guo1; Su, ZG
2009-03-01
Source PublicationPROCESS BIOCHEMISTRY
ISSN1359-5113
Volume44Issue:3Pages:277-282
AbstractA new protein refolding technique based on the use of the non-charged detergent Triton X-100 immobilized to the cross-linked agarose gel Sepharose High Performance has been developed. The new solid phase was used in combination with soluble beta-cyclodextrin (beta-CD) to refold recombinant Green Fluorescent Protein fused to Tobacco Etch Virus protease (GFPTEVP) expressed as inclusion bodies in E. coli. Previous attempts to refold recombinant GFPTEVP by dilution had failed. In the new procedure a column packed with Triton X-100-coupled Sepharose High Performance was used to capture unfolded GFPTEVP followed by elution using an increasing beta-CD concentration gradient. The yield of properly refolded GFPTEVP was 46% at a protein concentration of 380 mu g/ml. In contrast, dilution refolding of GFPTEVP at 200 mu g/ml refolding buffer resulted in only 4.7% of native protein. (C) 2008 Elsevier Ltd. All rights reserved.
KeywordImmobilized Triton X-100 Sepharose High Performance Refolding Green Fluorescent Protein Hydrophobic Interaction Chromatography Artificial Chaperone Beta-cyclodextrin
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine ; Technology
DOI10.1016/j.procbio.2008.10.021
Indexed BySCI
Language英语
WOS KeywordHYDROPHOBIC INTERACTION CHROMATOGRAPHY ; ARTIFICIAL CHAPERONE ; ESCHERICHIA-COLI ; INCLUSION-BODIES ; RENATURATION ; DETERGENT
WOS Research AreaBiochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Engineering
WOS SubjectBiochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Engineering, Chemical
WOS IDWOS:000264148400004
Citation statistics
Cited Times:11[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Version出版稿
Identifierhttp://ir.ipe.ac.cn/handle/122111/6715
Collection生化工程国家重点实验室
Corresponding AuthorSu, ZG
Affiliation1.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100080, Peoples R China
2.Uppsala Univ, Biomed Ctr, Dept Surface Biotechnol, SE-75123 Uppsala, Sweden
Recommended Citation
GB/T 7714
Li, Jing Jing,Wang, Ai-Qing,Janson, Jan-Christer,et al. Immobilized Triton X-100-assisted refolding of Green Fluorescent Protein-Tobacco Etch Virus protease fusion protein using beta-cyclodextrin as the eluent[J]. PROCESS BIOCHEMISTRY,2009,44(3):277-282.
APA Li, Jing Jing.,Wang, Ai-Qing.,Janson, Jan-Christer.,Ballagi, Andras.,Chen, Jing.,...&Su, ZG.(2009).Immobilized Triton X-100-assisted refolding of Green Fluorescent Protein-Tobacco Etch Virus protease fusion protein using beta-cyclodextrin as the eluent.PROCESS BIOCHEMISTRY,44(3),277-282.
MLA Li, Jing Jing,et al."Immobilized Triton X-100-assisted refolding of Green Fluorescent Protein-Tobacco Etch Virus protease fusion protein using beta-cyclodextrin as the eluent".PROCESS BIOCHEMISTRY 44.3(2009):277-282.
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