CAS OpenIR  > 研究所(批量导入)
Membrane combined with hydrophilic macromolecules enhances protein refolding at high concentration
Alternative TitleProcess Biochem.
Zhao, Dawei1,2; Liu, Yongdong1; Wang, Yinjue1; Li, Xiunan1; Wang, Qingqing1,2; Su, Zhiguo1
2014-07-01
Source PublicationPROCESS BIOCHEMISTRY
ISSN1359-5113
Volume49Issue:7Pages:1129-1134
AbstractMembrane technology is important to the development of modern biotechnology. It has the potential to efficiently refold protein at high concentration that is still a challenge for pharmaceutical protein produced from inclusion bodies. This paper dealt with the application of a polysulfone hollow fiber membrane to protein refolding using recombinant human granulocyte colony-stimulating factor (rhG-CSF) as a model protein. Compared with dilution refolding at protein concentration of 1.0 mg/mL, the crossflow membrane system led to a 16% increase in soluble protein recovery, and a 3.3-fold increase in specific bioactivity. Addition of PEG 6 K at 2 g/L could further improve the soluble protein recovery up to 57%, the specific bioactivity up to 2.2 x 10(8) IU/mL. Addition of dextran at 5 g/L could increase the soluble protein recovery up to 63.6%, the specific bioactivity up to 2.30 x 10(8) IU/mL. By gently and gradually removing denaturant, ultrafiltration membrane system was demonstrated to be very helpful for protein refolding at high concentration. Combining with hydrophilic macromolecular of PEG or dextran could further increase its efficiency. PEG was able to promote the refolding intermediate of rhG-CSF to transfer into the native structure; whereas dextran could enhance protein refolding mainly by weakening shear stress-induced protein aggregation. (C) 2014 Elsevier Ltd. All rights reserved.; Membrane technology is important to the development of modern biotechnology. It has the potential to efficiently refold protein at high concentration that is still a challenge for pharmaceutical protein produced from inclusion bodies. This paper dealt with the application of a polysulfone hollow fiber membrane to protein refolding using recombinant human granulocyte colony-stimulating factor (rhG-CSF) as a model protein. Compared with dilution refolding at protein concentration of 1.0 mg/mL, the crossflow membrane system led to a 16% increase in soluble protein recovery, and a 3.3-fold increase in specific bioactivity. Addition of PEG 6 K at 2 g/L could further improve the soluble protein recovery up to 57%, the specific bioactivity up to 2.2 x 10(8) IU/mL. Addition of dextran at 5 g/L could increase the soluble protein recovery up to 63.6%, the specific bioactivity up to 2.30 x 10(8) IU/mL. By gently and gradually removing denaturant, ultrafiltration membrane system was demonstrated to be very helpful for protein refolding at high concentration. Combining with hydrophilic macromolecular of PEG or dextran could further increase its efficiency. PEG was able to promote the refolding intermediate of rhG-CSF to transfer into the native structure; whereas dextran could enhance protein refolding mainly by weakening shear stress-induced protein aggregation. (C) 2014 Elsevier Ltd. All rights reserved.
KeywordRefolding Ultrafiltration Rhg-csf Inclusion Bodies Polyethylene Glycol Dextran
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine ; Technology
DOI10.1016/j.procbio.2014.03.010
URL查看原文
Indexed BySCI
Language英语
WOS KeywordHYDROPHOBIC INTERACTION CHROMATOGRAPHY ; ESCHERICHIA-COLI ; INCLUSION-BODIES ; HIGH-PRESSURE ; THERAPEUTIC PROTEINS ; RECOMBINANT PROTEINS ; IN-VITRO ; LYSOZYME ; AGGREGATION ; CELLS
WOS Research AreaBiochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Engineering
WOS SubjectBiochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Engineering, Chemical
WOS IDWOS:000338605500010
Citation statistics
Cited Times:2[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Version出版稿
Identifierhttp://ir.ipe.ac.cn/handle/122111/11048
Collection研究所(批量导入)
Affiliation1.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China
2.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China
Recommended Citation
GB/T 7714
Zhao, Dawei,Liu, Yongdong,Wang, Yinjue,et al. Membrane combined with hydrophilic macromolecules enhances protein refolding at high concentration[J]. PROCESS BIOCHEMISTRY,2014,49(7):1129-1134.
APA Zhao, Dawei,Liu, Yongdong,Wang, Yinjue,Li, Xiunan,Wang, Qingqing,&Su, Zhiguo.(2014).Membrane combined with hydrophilic macromolecules enhances protein refolding at high concentration.PROCESS BIOCHEMISTRY,49(7),1129-1134.
MLA Zhao, Dawei,et al."Membrane combined with hydrophilic macromolecules enhances protein refolding at high concentration".PROCESS BIOCHEMISTRY 49.7(2014):1129-1134.
Files in This Item:
File Name/Size DocType Version Access License
Membrane combined wi(1589KB) 限制开放CC BY-NC-SAApplication Full Text
Related Services
Recommend this item
Bookmark
Usage statistics
Export to Endnote
Google Scholar
Similar articles in Google Scholar
[Zhao, Dawei]'s Articles
[Liu, Yongdong]'s Articles
[Wang, Yinjue]'s Articles
Baidu academic
Similar articles in Baidu academic
[Zhao, Dawei]'s Articles
[Liu, Yongdong]'s Articles
[Wang, Yinjue]'s Articles
Bing Scholar
Similar articles in Bing Scholar
[Zhao, Dawei]'s Articles
[Liu, Yongdong]'s Articles
[Wang, Yinjue]'s Articles
Terms of Use
No data!
Social Bookmark/Share
All comments (0)
No comment.
 

Items in the repository are protected by copyright, with all rights reserved, unless otherwise indicated.