Design and Functional Investigation of a Novel Anti-coagulative Fusion Protein by Hirudin With a Recognizing Sequence of FXa
- VernacularTitle:新型水蛭素嵌合抗栓剂的构建表达与功能研究
- Author:
Jin-Yang NIU
;
Chun-Na DONG
;
Ji-De JIN
;
Bing-Xing SHI
;
Zu-Ze WU
;
- Publication Type:Journal Article
- Keywords:
Hirudin Bleeding side effect Thrombin Factor Xa
- From:
China Biotechnology
2006;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Hirudin (HV) is known as the most potent and specific inhibitor of thrombin. Although hirudin has many advantages , it has the bleeding side effect and this is the great shortage of hiudin for clinical application. In order to alleviate bleeding side effect of hirudin, fusion protein, named as FHV (fusion hirudin linked with FXa recognition peptide) was designed. The fusion protein gene ( fhv) was cloned into plasmid pPIC9K. FHV engineered Pichia pastoris containing high copies was chosen for fermentation and purification at 30 L fermentor scale, finally, FHV with purity of above 97% was obtained. To investigate the function of FHV in vivo, mouse tail thrombosis model was used. In the mice thrombus tail model induced by carrageenan, FHV decreased the length of tail thrombus significantly, similar to that of HV control, and had no obvious effects on the TT, PT and APTT. In conclusion, FHV is constructed and expressed in yeast. FHV fusion proteins is obtained by fermentation and purification. FHV has antithrombotic effects not influencing IT, PT and APTT after administration immediately in animal models. Therefore, FHV is a promising anticoagulant and antithrombotic drug.
