On-colunm refolding of consensus interferon at high concentration with guanidine-hydrochloride and polyethylene glycol gradients
文献类型：期刊论文
| 作者 | Wang, FW; Liu, YD; Li, JJ; Ma, GH; Su, ZG |
| 刊名 | JOURNAL OF CHROMATOGRAPHY A
 |
| 出版日期 | 2006-05-19 |
| 卷号 | 1115期号:1-2页码:72-80 |
| 关键词 | refolding protein hydrophobic interaction chromatography guanidine-HCl polyethylene glycol |
| ISSN号 | 0021-9673 |
| 其他题名 | J. Chromatogr. A |
| 中文摘要 | Dilution refolding of consensus interferon (C-IFN) had a limit on final concentration not exceeding 0.1 mg ml(-1) in order to achieve specific activity of 2.2 x 10(8) U mg(-1). Addition of polyethylence glycol (PEG) only gave a marginal improvement on the specific activity. Hydrophobic interaction chromatography (HIC) was tried but a simple step-wise elution could not refold the protein. Successful refolding was achieved by gradient elution with the decreasing of guanidine-hydrochloride (guanidine-HCl) concentration. The column was packed with a commercially available HIC medium that was designed for protein separation. Polyethylene glycol was found to possess better effect on the column than in the dilution for promotion of correct refolding, especially in gradient mode. A novel dual-gradient strategy, consisting of decreasing guanidine-HCl concentration and increasing PEG concentration, was developed to enhance the refolding yield. Denatured C-IFN was allowed to adsorb and elute from the HIC column through a gradually changed solution environment. Compared with dilution refolding, the gradient HIC process, in the presence of PEG, gave about 2.6-folds of increase in specific activity, 30% increase in soluble protein recovery. Partial purification was also achieved simultaneously. (c) 2006 Elsevier B.V. All rights reserved. |
| 英文摘要 | Dilution refolding of consensus interferon (C-IFN) had a limit on final concentration not exceeding 0.1 mg ml(-1) in order to achieve specific activity of 2.2 x 10(8) U mg(-1). Addition of polyethylence glycol (PEG) only gave a marginal improvement on the specific activity. Hydrophobic interaction chromatography (HIC) was tried but a simple step-wise elution could not refold the protein. Successful refolding was achieved by gradient elution with the decreasing of guanidine-hydrochloride (guanidine-HCl) concentration. The column was packed with a commercially available HIC medium that was designed for protein separation. Polyethylene glycol was found to possess better effect on the column than in the dilution for promotion of correct refolding, especially in gradient mode. A novel dual-gradient strategy, consisting of decreasing guanidine-HCl concentration and increasing PEG concentration, was developed to enhance the refolding yield. Denatured C-IFN was allowed to adsorb and elute from the HIC column through a gradually changed solution environment. Compared with dilution refolding, the gradient HIC process, in the presence of PEG, gave about 2.6-folds of increase in specific activity, 30% increase in soluble protein recovery. Partial purification was also achieved simultaneously. (c) 2006 Elsevier B.V. All rights reserved. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine ; Physical Sciences |
| 类目[WOS] | Biochemical Research Methods ; Chemistry, Analytical |
| 研究领域[WOS] | Biochemistry & Molecular Biology ; Chemistry |
| 关键词[WOS] | HYDROPHOBIC INTERACTION CHROMATOGRAPHY ; ESCHERICHIA-COLI ; STRUCTURAL CHARACTERIZATION ; DENATURED/REDUCED LYSOZYME ; INCLUSION-BODIES ; PROTEINS ; ALPHA ; RENATURATION ; MODEL |
| 收录类别 | SCI |
| 原文出处 | |
| 语种 | 英语 |
| WOS记录号 | WOS:000237481400007 |
| 公开日期 | 2013-10-24 |
| 版本 | 出版稿 |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/4032]  |
| 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, State Key Lab Biochem Engn, Beijing 100080, Peoples R China 2.Dalian Univ Technol, Dept Biosci & Biotechnol, Dalian 116024, Peoples R China 3.Dalian Med Univ, Clin Coll 2, Dalian 116027, Peoples R China |
| 推荐引用方式 GB/T 7714 | Wang, FW,Liu, YD,Li, JJ,et al. On-colunm refolding of consensus interferon at high concentration with guanidine-hydrochloride and polyethylene glycol gradients[J]. JOURNAL OF CHROMATOGRAPHY A,2006,1115(1-2):72-80. |
| APA | Wang, FW,Liu, YD,Li, JJ,Ma, GH,&Su, ZG.(2006).On-colunm refolding of consensus interferon at high concentration with guanidine-hydrochloride and polyethylene glycol gradients.JOURNAL OF CHROMATOGRAPHY A,1115(1-2),72-80. |
| MLA | Wang, FW,et al."On-colunm refolding of consensus interferon at high concentration with guanidine-hydrochloride and polyethylene glycol gradients".JOURNAL OF CHROMATOGRAPHY A 1115.1-2(2006):72-80. |
入库方式： OAI收割
来源：过程工程研究所
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