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J. Microbiol. Biotechnol. 2008; 18(1): 183-188

Published online January 28, 2008

Copyright © The Korean Society for Microbiology and Biotechnology.

Overproduction of Recombinant Human VEGF (Vascular Endothelial Growth Factor) in Chinese Hamster Ovary Cells

Lee, Seong-Baek 2, 3, Jeong Soo Park 2, Seunghee Lee 2, Junho Park 2, Sungryul Yu 1, Heechan Kim 1, Dongjun Kim 2, Tae Ho Byun 2, Kwanghee Baek 1, Young-Joon Ahn 3 and Jaeseung Yoon 2*

1Graduate School of Biotechnology, Kyung Hee University, Yongin 449-701, Korea, 2PanGen Biotech Inc., 5F NKBio Bldg., Seongnam 462-807, Korea, 3School of Agricultural Biotechnology, Seoul National University, Seoul 151-921, Korea

Abstract

Vascular endothelial growth factors (VEGFs) are a family of proteins that mediate angiogenesis. $VEGF_{165}$ is a VEGF-A isoform and has been extensively studied owing to its potential use in therapeutic angiogenesis. This study established Chinese hamster ovary (CHO) cells overexpressing recombinant human $VEGF_{165}$ $(rhVEGF_{165})$ protein. The production rate of the established CHO cells was over 80mg/l of $rhVEGF_{165}$ protein from a 7-day batch culture process using a 7.5-l bioreactor with a 5-l working volume and serum-free medium. The $rhVEGF_{165}$ protein was purified to homogeneity from the culture supernatant using a two-step chromatographic procedure that resulted in a 48% recovery rate. The purified $rhVEGF_{165}$ protein was a glycosylated homodimeric protein with a higher molecular weight (MW) than the protein expressed from insect cells, suggesting that the glycosylation of the $rhVEGF_{165}$ protein in CHO cells differed from that in insect cells. The purified $rhVEGF_{165}$ protein in this study was functionally active with a half-maximal effective concentration of 3.8ng/ml and specific activity of $2.5{\times}10^5U/mg$.

Keywords: recombinant protein expression, stable cell line, bioreactor process, VEGF, Chinese hamster ovary (CHO) cell