| 摘要 |
PURPOSE: Preparation method of mass production of biologically active EPO from recombinant plasmid in CHO(chinese hamster ovary) cell is provided. CONSTITUTION: The invention is disclosed a mass production method from recombinant plasmid pDEP-173 consisted of zeocin resistance gene, EPO gene, isolated from human fetal liver cDNA library, under control of SRα promoter, AmvRNA4 enhancer, DHFR gene, and BGH polyadenylation signal. The plasmid is transfected into CHO cell and cultured with roller bottle in serum free medium. EPO is induced with n-butyric acid or methotrexate. Overexpressed EPO is purified with hydrophobic chromatography and anion exchange chromatography. The purified EPO only having N-acetyl neuraminic acid as N-terminal end of sialic acid has molecular weight of 34,000-47,000 and shows 7-8 isomer bands in isoelectric focusing around pH 3.7-5.2.
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