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| 重组质粒pUDK-HGF 的中试纯化工艺 | |
| 引用本文： | 胡春生,王延亮,卢育新,程晓晨,刘琳,张通,张庆林.重组质粒pUDK-HGF 的中试纯化工艺[J].生物工程学报,2011,27(2):247-252. |
| 作者姓名： | 胡春生 王延亮 卢育新 程晓晨 刘琳 张通 张庆林 |
| 作者单位： | 1. 军事医学科学院放射与辐射医学研究所,北京,100850;内蒙古工业大学化工学院,呼和浩特,010051 2. 军事医学科学院放射与辐射医学研究所,北京,100850 3. 内蒙古工业大学化工学院,呼和浩特,010051 |
| 基金项目： | 国家高技术研究发展计划 (863计划) (No. 2001AA217061) 资助。 |
| 摘 要： | pUDK-HGF是携带人肝细胞生长因子的裸质粒，目前已进入I期临床试验，因此需要大量符合药学规格的质粒DNA。文中建立了pUDK-HGF中试规模纯化制备的新工艺。流程包括：发酵、离心收获菌体、碱裂解、超滤浓缩碱裂解液、Sephacryl S-1000层析除去RNA并更换缓冲液、plasmidselect捕获超螺旋质粒DNA、琼脂糖凝胶6BFF除盐。新工艺可获得浓度为2.0 mg/mL、纯度在1.70以上的裸质粒原液，符合相关质量标准，并避免使用动物源性的酶及有毒试剂。 |
| 关 键 词： | 重组质粒，纯化，质量检测，肝细胞生长因子 |
| 收稿时间： | 6/7/2010 12:00:00 AM |
| 修稿时间： | 2010/8/25 0:00:00 |
Pilot-scale production of recombinant plasmid pUDK-HGF |
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| Chunsheng Hu,Yanliang Wang,Yuxin Lu,Xiaochen Cheng,Lin Liu,Tong Zhang and Qinglin Zhang.Pilot-scale production of recombinant plasmid pUDK-HGF[J].Chinese Journal of Biotechnology,2011,27(2):247-252. | |
| Authors: | Chunsheng Hu Yanliang Wang Yuxin Lu Xiaochen Cheng Lin Liu Tong Zhang and Qinglin Zhang |
| Institution: | Beijing Institute of Radiation Medicine, Beijing 100850, China; College of Chemical Engineering, Inner Mongolia University of Technology, Huhhot 010051, China;Beijing Institute of Radiation Medicine, Beijing 100850, China;Beijing Institute of Radiation Medicine, Beijing 100850, China;Beijing Institute of Radiation Medicine, Beijing 100850, China;Beijing Institute of Radiation Medicine, Beijing 100850, China;College of Chemical Engineering, Inner Mongolia University of Technology, Huhhot 010051, China;Beijing Institute of Radiation Medicine, Beijing 100850, China |
| Abstract: | pUDK-HGF, the recombinant plasmid DNA encoding human hepatocyte growth factor (HGF), can treat ischaemic disease. A great quantity of pharmaceutical pUDK-HGF is needed. A pilot-scale production process of pUDK-HGF was established based on a new chromatographic media (plasmidselect), including fermentation, cell harvesting, alkaline lysis, ultrafiltration, RNA removing and buffer exchanging on Sephacryl S-1000, capturing supercoiled plasmid DNA with plasmidselect, and removing the salt with Sepharose 6BFF. The process does not use RNase enzyme and toxic solvents. |
| Keywords: | recombinant plasmid purification hepatocyte growth factor |
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