Membrane combined with hydrophilic macromolecules enhances protein refolding at high concentration
文献类型：期刊论文
| 作者 | Zhao, Dawei1,2; Liu, Yongdong1; Wang, Yinjue1; Li, Xiunan1; Wang, Qingqing1,2; Su, Zhiguo1 |
| 刊名 | PROCESS BIOCHEMISTRY
 |
| 出版日期 | 2014-07-01 |
| 卷号 | 49期号:7页码:1129-1134 |
| 关键词 | Refolding Ultrafiltration rhG-CSF Inclusion bodies Polyethylene glycol Dextran |
| ISSN号 | 1359-5113 |
| 其他题名 | Process Biochem. |
| 中文摘要 | 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. |
| 英文摘要 | 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. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine ; Technology |
| 类目[WOS] | Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Engineering, Chemical |
| 研究领域[WOS] | Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Engineering |
| 关键词[WOS] | HYDROPHOBIC INTERACTION CHROMATOGRAPHY ; ESCHERICHIA-COLI ; INCLUSION-BODIES ; HIGH-PRESSURE ; THERAPEUTIC PROTEINS ; RECOMBINANT PROTEINS ; IN-VITRO ; LYSOZYME ; AGGREGATION ; CELLS |
| 收录类别 | SCI |
| 原文出处 | |
| 语种 | 英语 |
| WOS记录号 | WOS:000338605500010 |
| 公开日期 | 2014-08-28 |
| 版本 | 出版稿 |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/11048]  |
| 专题 | 过程工程研究所_研究所（批量导入） |
| 作者单位 | 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 |
| 推荐引用方式 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. |
入库方式： OAI收割
来源：过程工程研究所
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