Refolding and purification of recombinant human VEGF-121 expressed as inclusion bodies in Escherichia coli.
- Author:
Zhi-ming HU
1
;
Li MA
;
Ming-qian ZHOU
;
Ji-min GAO
;
Xiao-ning WANG
Author Information
1. Institute of Molecular Immunology, Southern Medical University, Guangzhou 510515, China. hzm@fimmu.com
- Publication Type:Journal Article
- MeSH:
Cell Line;
Cell Proliferation;
drug effects;
Dose-Response Relationship, Drug;
Endothelial Cells;
cytology;
drug effects;
Escherichia coli;
genetics;
metabolism;
Humans;
Inclusion Bodies;
metabolism;
Protein Folding;
Recombinant Proteins;
biosynthesis;
chemistry;
isolation & purification;
Vascular Endothelial Growth Factor A;
biosynthesis;
genetics;
pharmacology
- From:
Journal of Southern Medical University
2006;26(8):1083-1086
- CountryChina
- Language:English
-
Abstract:
Vascular endothelial growth factor 121 (VEGF(121)) was expressed as inclusion bodies by recombinant Escherichia coli. High concentrations of both biomass (46 g dry cell/L) and VEGF(121) inclusion bodies (4.5 g/L) were obtained by applying a high-cell-density culture. After the inclusion bodies were washed and dissolved, VEGF(121) was refolded at 0.2 mg/ml by ultrafiltration in refolding buffer with a yield of 81%. Renatured VEGF(121) was purified by anion chromatography and Sephacry S-100 chromatography with purity higher than 95% and final purification yield of 31%. The purified VEGF(121) could stimulate the proliferation of human umbilical vein endothelial cells as demonstrated by a biological activity assay.
