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| Prokaryotic Expression of VEGF165 and Preliminary Screening of Anti-VEGF Hybridoma Cell Lines |
| BAO Xin-chen1, LI Zi1, GAO Xiang-dong1, LU Xiao-dong2, XU Chen2 |
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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Received: 25 April 2011
Published: 25 September 2011
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