Overproduction of recombinant human VEGF (vascular endothelial growth factor) in Chinese hamster ovary cells.
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 x 105 U/mg.
Lee
Seong-Baek
SB
PanGen Biotech Inc., 5F NKBio Bldg., Seongnam 462-807, Korea.
Park
Jeong Soo
JS
Lee
Seunghee
S
Park
Junho
J
Yu
Sungryul
S
Kim
Heechan
H
Kim
Dongjun
D
Byun
Tae Ho
TH
Baek
Kwanghee
K
Ahn
Young-Joon
YJ
Yoon
Jaeseung
J
D000818:Animals; D019149:Bioreactors; D001709:Biotechnology / Q000379:methods*; D016466:CHO Cells / Q000378:metabolism*; D002478:Cells, Cultured; D006224:Cricetinae; D003412:Cricetulus; D003470:Culture Media; D042783:Endothelial Cells; D004730:Endothelium, Vascular / Q000166:cytology; D006801:Humans; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D014471:Umbilical Veins; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology
D003470:Culture Media; D011994:Recombinant Proteins; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A