Reproducible preparation and effective separation of PEGylated recombinant human granulocyte colony-stimulating factor with novel "PEG-pellet" PEGylation mode and ion-exchange chromatography
文献类型：期刊论文
| 作者 | Yun, Q; Yang, RE; Chen, T; Bi, JX; Ma, GH; Su, ZG |
| 刊名 | JOURNAL OF BIOTECHNOLOGY
 |
| 出版日期 | 2005-07-21 |
| 卷号 | 118期号:1页码:67-74 |
| ISSN号 | 0168-1656 |
| 关键词 | chromatography PEGylation poly(ethylene glycol) protein rhG-CSF |
| 其他题名 | J. Biotechnol. |
| 中文摘要 | A novel preparation for polyethylene glycol (PEG) derivatives and chromatographic separation procedure of the PEGylated recombinant human granulocyte colony-stimulating factor (rhG-CSF) were designed to evaluate the reproducibility and scalability at large laboratory-scale level. The new "PEG-pellet" PEGylation mode was successfully applied to control the pH fluctuation during the conjugation reaction, a general problem in traditional liquid-phase conjugation mode. Moreover, two consecutive ion-exchange chromatography steps were successfully used to separate and purify the PEGylated rhG-CSF. Cation-exchange chromatography was firstly applied to separate PEGylated rhG-CSF from intact rhG-CSF, followed by anion-exchange chromatography to obtain individual PEG-rhG-CSF species (mono-, di- and tri-PEGylated rhG-CSF) and remove the excess free PEG. Furthermore, the molecular weight of individual PEGylated rhG-CSF was identified by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry and SDS-PAGE, and cell proliferation activity in vitro was detected by MTT assay using NFS-60 cell. (c) 2005 Elsevier B.V. All rights reserved. |
| 英文摘要 | A novel preparation for polyethylene glycol (PEG) derivatives and chromatographic separation procedure of the PEGylated recombinant human granulocyte colony-stimulating factor (rhG-CSF) were designed to evaluate the reproducibility and scalability at large laboratory-scale level. The new "PEG-pellet" PEGylation mode was successfully applied to control the pH fluctuation during the conjugation reaction, a general problem in traditional liquid-phase conjugation mode. Moreover, two consecutive ion-exchange chromatography steps were successfully used to separate and purify the PEGylated rhG-CSF. Cation-exchange chromatography was firstly applied to separate PEGylated rhG-CSF from intact rhG-CSF, followed by anion-exchange chromatography to obtain individual PEG-rhG-CSF species (mono-, di- and tri-PEGylated rhG-CSF) and remove the excess free PEG. Furthermore, the molecular weight of individual PEGylated rhG-CSF was identified by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry and SDS-PAGE, and cell proliferation activity in vitro was detected by MTT assay using NFS-60 cell. (c) 2005 Elsevier B.V. All rights reserved. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 类目[WOS] | Biotechnology & Applied Microbiology |
| 研究领域[WOS] | Biotechnology & Applied Microbiology |
| 关键词[WOS] | POLYETHYLENE-GLYCOL ; PROTEINS ; CONJUGATION ; ASSAY ; MASS |
| 收录类别 | SCI |
| 原文出处 | |
| 语种 | 英语 |
| WOS记录号 | WOS:000230328500007 |
| 公开日期 | 2013-10-25 |
| 版本 | 出版稿 |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/4246]  |
| 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 100080, Peoples R China 2.Beijing Univ Chem Technol, Sch Chem Engn, Beijing 100029, Peoples R China 3.Univ Sci & Technol Beijing, Sch Civil & Environm Engn, Beijing 100083, Peoples R China |
| 推荐引用方式 GB/T 7714 | Yun, Q,Yang, RE,Chen, T,et al. Reproducible preparation and effective separation of PEGylated recombinant human granulocyte colony-stimulating factor with novel "PEG-pellet" PEGylation mode and ion-exchange chromatography[J]. JOURNAL OF BIOTECHNOLOGY,2005,118(1):67-74. |
| APA | Yun, Q,Yang, RE,Chen, T,Bi, JX,Ma, GH,&Su, ZG.(2005).Reproducible preparation and effective separation of PEGylated recombinant human granulocyte colony-stimulating factor with novel "PEG-pellet" PEGylation mode and ion-exchange chromatography.JOURNAL OF BIOTECHNOLOGY,118(1),67-74. |
| MLA | Yun, Q,et al."Reproducible preparation and effective separation of PEGylated recombinant human granulocyte colony-stimulating factor with novel "PEG-pellet" PEGylation mode and ion-exchange chromatography".JOURNAL OF BIOTECHNOLOGY 118.1(2005):67-74. |
入库方式： OAI收割
来源：过程工程研究所
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