標題: | Prokaryotic expression, refolding, and purification of functional human vascular endothelial growth factor isoform 165: Purification procedures and refolding conditions revisited |
作者: | Lee, I-Liang Li, Pei-Shan Yu, Wei-Lin Shen, Hsin-Hsin 生物科技學系 Department of Biological Science and Technology |
關鍵字: | Expression;Purification;Refolding;Recombinant VEGF protein;Angiogenesis;Untagged protein |
公開日期: | 1-三月-2011 |
摘要: | Human vascular endothelial growth factor isoform 165 (VEGF165) is the first known member belonging to the VEGF protein family that plays a critical role in new blood vessel formation in vivo. This study presents a new protocol with optimized conditions for rapidly producing untagged recombinant and biological active human VEGF165 (rhVEGF165) using Escherichia coli cells. Protein was isolated from inclusion bodies, purified by gel filtration and ion exchange chromatography, and subjected to protein refolding and renaturation. The biological activity of rhVEGF165 is comparable with VEFG from eukaryotic source according to human umbilical vein endothelial cells (HUVEC) proliferation assay. Therefore, the present procedures provide a fast and easy way to produce this therapeutic protein. (C) 2010 Published by Elsevier Inc. |
URI: | http://dx.doi.org/10.1016/j.pep.2010.08.014 http://hdl.handle.net/11536/9266 |
ISSN: | 1046-5928 |
DOI: | 10.1016/j.pep.2010.08.014 |
期刊: | PROTEIN EXPRESSION AND PURIFICATION |
Volume: | 76 |
Issue: | 1 |
起始頁: | 54 |
結束頁: | 58 |
顯示於類別: | 期刊論文 |