| 疏水作用层析在重组大肠杆菌表达的rProEMAPⅡ/P43蛋白纯化中的作用 |
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| 引用本文： | 魏健,刘大涛,谭小钉,储炬,庄英萍,张嗣良. 疏水作用层析在重组大肠杆菌表达的rProEMAPⅡ/P43蛋白纯化中的作用[J]. 中国生物制品学杂志, 2006, 19(1): 73-76 |
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| 作者姓名： | 魏健 刘大涛 谭小钉 储炬 庄英萍 张嗣良 |
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| 作者单位： | 华东理工大学生物反应器工程国家重点实验室 上海200237(魏健，储炬，庄英萍，张嗣良)，上海医药集团信谊药业药物研究所 上海201206(刘大涛，谭小钉) |
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| 基金项目： | 国家科技攻关项目;国家科技专项基金;上海市青年科技启明星计划 |
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| 摘 要： | 目的建立rProEMAPⅡ/P43蛋白的纯化工艺,明确疏水作用层析(HIC)在纯化中的作用。方法采用由疏水作用层析、离子交换层析、亲和肝素层析组成的工艺对rProEMAPⅡ/P43蛋白进行纯化,并与由离子交换层析和亲和肝素层析组成的工艺相比较,分别进行总蛋白含量测定、SDS-PAGE分析目的蛋白含量及纯度检测。结果在pH为7·4~7·5时,采用HIC方法纯化的蛋白较不采用HIC法回收率提高了28·4%,纯化后rProEMAPⅡ/P43蛋白纯度为92%。结论在rProEMAPⅡ/P43蛋白纯化过程中增加疏水作用层析,可以提高rProEMAPⅡ/P43蛋白收率。
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| 关 键 词： | rProEMAPⅡ/P43 HIC 回收率 纯度 |
| 收稿时间： | 2005-11-04 |
| 修稿时间： | 2005-11-04 |
Role of Hydrophobic Interaction Chromatography in Purification of rProEMAP Ⅱ/P43 Protein Expressed in E. coli |
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| WEI Jian , LIU Da-tao, TAN Xiao-ding, et al. Role of Hydrophobic Interaction Chromatography in Purification of rProEMAP Ⅱ/P43 Protein Expressed in E. coli[J]. Chinese Journal of Bilogicals, 2006, 19(1): 73-76 |
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| Authors: | WEI Jian LIU Da-tao TAN Xiao-ding et al |
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| Affiliation: | State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China |
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| Abstract: | Objective To develop a purification procedure of rProEMAPⅡ/P43.Methods The rProEMAPⅡ/P43 protein expressed in E.coli was purified by procedure 1,consisting of hydrophobic interaction chromatography,ion exchange chromatography and heparin affinity chromatography,and procedure 2,consisting of ion exchange chromatography and heparin affinity chromatography,respectively.The purified rProEMAPⅡ/P43 was determined for total protein content and analyzed for the content and purity of target protein by SDS-PAGE.Results Compared with that purified by procedure 2,the recovery rate of rProEMAPⅡ/P43 protein purified by procedure 1 at pH 7.4~7.5 increased by 28.4%,and its purity reached 92%.Conclusion The introduction of hydrophobic interaction chromatography into purification procedure increased the recovery rate of rProEMAPⅡ/P43 protein expressed in E.coli significantly. |
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| Keywords: | HIC |
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