Knowledge Management System Of Institute of process engineering,CAS
| Membrane combined with hydrophilic macromolecules enhances protein refolding at high concentration | |
| Alternative Title | Process Biochem. |
| Zhao, Dawei1,2; Liu, Yongdong1; Wang, Yinjue1; Li, Xiunan1; Wang, Qingqing1,2; Su, Zhiguo1 | |
| 2014-07-01 | |
| Source Publication | PROCESS BIOCHEMISTRY
 |
| ISSN | 1359-5113 |
| Volume | 49Issue:7Pages:1129-1134 |
| Abstract | 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.; 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. |
| Keyword | Refolding Ultrafiltration Rhg-csf Inclusion Bodies Polyethylene Glycol Dextran |
| Subtype | Article |
| WOS Headings | Science & Technology ; Life Sciences & Biomedicine ; Technology |
| DOI | 10.1016/j.procbio.2014.03.010 |
| URL | 查看原文 |
| Indexed By | SCI |
| Language | 英语 |
| WOS Keyword | HYDROPHOBIC INTERACTION CHROMATOGRAPHY ; ESCHERICHIA-COLI ; INCLUSION-BODIES ; HIGH-PRESSURE ; THERAPEUTIC PROTEINS ; RECOMBINANT PROTEINS ; IN-VITRO ; LYSOZYME ; AGGREGATION ; CELLS |
| WOS Research Area | Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Engineering |
| WOS Subject | Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Engineering, Chemical |
| WOS ID | WOS:000338605500010 |
| Citation statistics | |
| Document Type | 期刊论文 |
| Version | 出版稿 |
| Identifier | http://ir.ipe.ac.cn/handle/122111/11048 |
| Collection | 研究所（批量导入） |
| Affiliation | 1.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. |
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| Membrane combined wi（1589KB） | 限制开放 | CC BY-NC-SA | Application Full Text | |||
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