1.Stable expression of rhVEGF165 in Chinese hamster ovary cells.
Ganliang ZHANG ; Haifeng ZHANG ; Weihong NIAN ; Jiuru SUN
Chinese Journal of Biotechnology 2011;27(6):935-942
We established a stable Chinese hamster ovary (CHO-S) cell line for recombinant human VEGF165-expressing. We co-transfected GS-expression vector and rhVEGF165 expression plasmid into CHO-S cells, and selected the highest VEGF165-expressing clone as the working cell line to express VEGF165 protein. After 7-day fed-batch culture in a 5 L bioreactor and 3 steps chromatographic purification, we got the rhVEGF165 protein for series of binding and biological activity examination. The production was over 50 mg/L. The purified rhVEGF165 protein was functionally active with a half-maximal Human Umbilical Vein Endothelial Cells (HUVEC) growth-enhancing effect concentration of 1.94 ng/mL. It was slightly better than commercially available Escherichia coli expressing rhVEGF165. So we expressed successfully rhVEGF165 protein in high-level and obtained the fully active rhVEGF165 protein in large quantity.
Animals
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Bioreactors
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CHO Cells
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Cell Proliferation
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drug effects
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Cricetinae
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Cricetulus
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Human Umbilical Vein Endothelial Cells
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cytology
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Humans
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Recombinant Proteins
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biosynthesis
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genetics
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isolation & purification
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Transfection
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Vascular Endothelial Growth Factor A
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biosynthesis
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genetics