Expression, Purification of Human Endostatin in Pichia pastoris and the Detection of Its Anti-angiogenic Activity
- VernacularTitle:人血管内皮抑制素在毕赤酵母中的表达纯化及活性测定
- Author:
Xue-Jing YAO
;
Zhuang-Lin LI
;
Guo-Dong CHANG
;
Gui-Yong YUAN
;
Xue- YU
;
- Publication Type:Journal Article
- Keywords:
Endostatin Angiogenesis Pichia pastoris Purification
- From:
China Biotechnology
2006;0(04):-
- CountryChina
- Language:Chinese
-
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.
