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KMID : 0545120080180010183
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Journal of Microbiology and Biotechnology
2008 Volume.18 No. 1 p.183 ~ p.187
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Overproduction of Recombinant Human VEGF (Vascular Endothelial Growth Factor) in Chinese Hamster Ovary Cells
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Lee Seong-Baek
Park Jeong-Soo
Lee Seung-Hee
Park Jun-Ho
Yu Sung-Ryul
Kim Hee-Chan
Kim Dong-Jun
Byun Tae-Ho
Baek Kwang-Hee
Ahn Young-Joon
Yoon Jae-Seung
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Abstract
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Vascular endothelial growth factors (VEGFs) are a family of proteins that mediate angiogenesis. VEGF165 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 VEGF165 (rhVEGF165) protein. The production rate of the established CHO cells was over 80 mg/l of rhVEGF165 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 rhVEGF165 protein was purified to homogeneity from the culture supernatant using a two-step chromatographic procedure that resulted in a 48% recovery rate. The purified rhVEGF165 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 rhVEGF165 protein in CHO cells differed from that in insect cells. The purified rhVEGF165 protein in this study was functionally active with a half-maximal effective concentration of 3.8 ng/ ml and specific activity of 2.5×105 U/mg.
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KEYWORD
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VEGF, Chinese hamster ovary (CHO) cell, recombinant protein expression, stable cell line, bioreactor process
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