Expression, Purification of Human Endostatin in Pichia pastoris and the Detection of Its Anti-angiogenic Activity / 中国生物工程杂志

ABSTRACT

Endostatin is a potent, endogenous inhibitor of angiogenesis, which corresponds to the C-terminal fragment of collagen ⅩⅧ. The human endostatin gene was amplified from a human fetal liver cDNA library by means of PCR and was cloned into pPIC9K vector. Soluble endostatin was superexpressed in Pichia pastoris (50.5mg/L) . The recombinant protein was purified by cation exchange chromatography with the final yield of more than 95%. Western blotting analysis showed positive immunoreactivity to the expressed product. Recombinant endostatin was able to suppress the angiogenesis on the CAMs. Also, the endostatin inhibited specifically of the migration of HMECs stimulated by bFGF, with EC50 being about 0.4 ?g/ml.