Purification of Recombinant Fusion Protein Staphylokinase-Hirudin Expressed by Escherichia coli and Analysis of its Dimer
- VernacularTitle:大肠杆菌表达的重组葡激酶-水蛭素融合蛋白的分离纯化及其二聚体分析
- Author:
Gen-Shen ZHONG
;
Ai-Ping YU
;
Ji-De JIN
;
Zhong-Hua JIANG
;
Zu-Ze WU
;
- Publication Type:Journal Article
- Keywords:
Staphylokinase Hirudin rSFH Purification Dimerization
- From:
China Biotechnology
2006;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
The recombinant fusion protein staphylokinase-hirudin(rSFH) was purified from the high density-fermented engineered E.coli by means of ion-exchange chromatography (IEC) and gel filtration (GF). The purity of rSFH reached to more than 98% determined by RP-HPLC and SDS-PAGE, and the yield was up to 0.7g per liter of fermentation broth. The analysis of homologous dimmer of rSFH appeared during the purification and calculation of the surface hydrophobic area had been carried out by means of hydrophobic chromatography and MALD-TOF. The influence of sodium chloride and temperature on the behavior of rSFH reversible dimerization was analyzed by high performance sized- exclusive chromatography(HPSEC). It is concluded that the hydrophobic interaction played an important role in the reversible dimerization of rSFH.
