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| The Antithrombin Activity Detection of EH in Vitro |
| QIN Xiao-yong1, YU Ai-ping2, WANG Wen-wen3, BI Jian-jin2, GUO Xiao-jun1, YUAN Hong-shui1, WU Zu-ze2, JIN Ji-de2, ZHU Bao-cheng1 |
1. College of Life Science, Agricultural University of Hebei, Baoding 071001, China;
2. Department of Experimental Hematology, Beijing Institute of Radiation Medicine, Beijing 100850, China;
3. School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China |
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Abstract Recombinant protein EH originates from hirudin by linking three amino acids to its N-terminal. The three amino acids are recognized and cut by coagulate factor Xa (FXa) and XIa (FXIa), that will release the antithrombin activity of hirudin.The cleavage efficiency of FXa to EH was compared under different conditions, incuding emzymolytic time and temperature, the molar ratio of FXa and EH, and reaction solution. Results showed that the antithrombin activity of EH increased proportionally, under certain conditions, as the reaction time and temperature increased, Moreover, the FXa cleavage activity is enhanced when the molar ratio of FXa and EH was increased. It seems that saline is more suitable reaction solutions for FXa to cut EH. So the optimal cleavage conditions for EH with FXa in vitro is that EH was reacted with FXa at 37℃ for 6 hours in saline solution with the molar ratio (1 ∶180) of FXa and EH. A practical and reproducible method for the activity determination and quality control of EH in vitro was developed.
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Received: 20 January 2011
Published: 27 May 2011
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Cite this article:
QIN Xiao-yong, YU Ai-ping, WANG Wen-wen, BI Jian-jin, GUO Xiao-jun, YUAN Hong-shui, WU Zu-ze, JIN Ji-de, ZHU Bao-cheng. The Antithrombin Activity Detection of EH in Vitro. China Biotechnology, 2011, 31(5): 108-112.
URL:
https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2011/V31/I5/108
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