Refolding and purification of recombinant human VEGF-121 expressed as inclusion bodies in Escherichia coli.
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.
Hu
Zhi-ming
ZM
Institute of Molecular Immunology, Southern Medical University, Guangzhou 510515, China. hzm@fimmu.com
Ma
Li
L
Zhou
Ming-qian
MQ
Gao
Ji-min
JM
Wang
Xiao-ning
XN
D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D002479:Inclusion Bodies / Q000378:metabolism*; D017510:Protein Folding; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology
D011994:Recombinant Proteins; D042461:Vascular Endothelial Growth Factor A