| 重组人VEGF165的表达纯化及单抗的初步筛选 |
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| 引用本文： | 包欣晨,李孜,高向东,陆小冬,徐晨.重组人VEGF165的表达纯化及单抗的初步筛选[J].中国生物工程杂志,2011,31(9):43-47. |
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| 作者姓名： | 包欣晨 李孜 高向东 陆小冬 徐晨 |
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| 作者单位： | 1. 中国药科大学生命科学与技术学院 南京 210009;
2. 北京三元基因工程有限公司 北京 102600 |
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| 摘 要： | 血管内皮细胞生长因子(vascular endothelial growth factor, VEGF)是抑制肿瘤生长和转移的重要靶点。为获得抗VEGF单抗细胞株,构建了rhVEGF165工程菌,并利用复合自动诱导获得高效表达。经纯化获得高纯度rhVEGF165蛋白,经检测具有促人脐静脉内皮细胞(human umbilical vein endothelial cells, HUVECs)增殖活性,其EC50为2.4ng/ml。免疫小鼠,获得了3株能稳定分泌抗VEGF单抗的杂交瘤细胞株,为开发VEGF治疗性单抗提供了重要基础。
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| 关 键 词： | 血管内皮细胞生长因子 原核表达 增殖活性 单克隆抗体 |
| 收稿时间： | 2011-04-25 |
| 修稿时间： | 2011-06-08 |
Prokaryotic Expression of VEGF165 and Preliminary Screening of Anti-VEGF Hybridoma Cell Lines |
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| BAO Xin-chen,LI Zi,GAO Xiang-dong,LU Xiao-dong,XU Chen.Prokaryotic Expression of VEGF165 and Preliminary Screening of Anti-VEGF Hybridoma Cell Lines[J].China Biotechnology,2011,31(9):43-47. |
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| Authors: | BAO Xin-chen LI Zi GAO Xiang-dong LU Xiao-dong XU Chen |
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| Institution: | 1 College of Life Science and Technology,China Pharmaceutical University,Nanjing 210009,China)(2 Beijing Tri-Prime Genetic Engineering Incorporated,Beijing 102600,China) |
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| Abstract: | Vascular endothelial growth factor (VEGF) plays a key role in physiological and pathological angiogenesis and is a potent and critical target as blocking tumor growth and metastasis. The process described in this report involves a complex auto-induced expression in Escherichia coli and a downstream purification process consisting of protein refolding and three chromatography steps in order to obtain the functional rhVEGF165. Biological activity of the purified 38kDa homodimer was verified by the induction of the proliferation of human umbilical vein endothelial cells (HUVECs). The EC50 for this effect was 2.4ng/ml. Finally, three Anti-VEGF hybridoma cell lines were obtained after immunization, fusion and preliminary screening. |
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| Keywords: | VEGF Prokaryotic expression Induction of the proliferation Monoclonal antibody |
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