Refolding and purification of recombinant human VEGF-121 expressed as inclusion bodies in Escherichia coli

Abstract

Abstract: Vascular endothelial growth factor 121 (VEGF121) was expressed as inclusion bodies by recombinant Escherichia coli. High concentrations of both biomass (46 g dry cell/L) and VEGF121 inclusion bodies (4.5 g/L) were obtained by applying a high-cell-density culture. After the inclusion bodies were washed and dissolved, VEGF121 was refolded at 0.2 mg/ml by ultrafiltration in refolding buffer with a yield of 81%. Renatured VEGF121 was purified by anion chromatography and Sephacry S-100 chromatography with purity higher than 95% and final purification yield of 31%. The purified VEGF121 could stimulate the proliferation of human umbilical vein endothelial cells as demonstrated by a biological activity assay.

CLC Number:

R392

HU Zhi-ming1, MA Li1, ZHOU Ming-qian1, GAO Ji-min1, WANG Xiao-ning1, 2 1Institute of Molecular Immunology, Southern Medical University, Guangzhou 510515, China; 2School of Biosciences & Bioengineering, South China University of Technology, Guangzhou 510641, China. Refolding and purification of recombinant human VEGF-121 expressed as inclusion bodies in Escherichia coli[J]. Journal of Southern Medical University, 2006, 26(08): 1083-1086.

References

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