title: Prokaryotic expression, refolding, and purification of functional human vascular endothelial growth factor isoform 165: purification procedures and refolding conditions revisited.
abstract: 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.
authors.affiliation: Department of Biological Science and Technology, National Chiao Tung University, Hsinchu 300, Taiwan.
authors.forename: I-Liang
authors.initials: IL
authors.lastname: Lee
authors.forename: Pei-Shan
authors.initials: PS
authors.lastname: Li
authors.forename: Wei-Lin
authors.initials: WL
authors.lastname: Yu
authors.forename: Hsin-Hsin
authors.initials: HH
authors.lastname: Shen
mesh_terms: D004926:Escherichia coli; D006801:Humans; D002479:Inclusion Bodies / Q000737:chemistry; D058849:Protein Refolding; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D011994:Recombinant Proteins; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A