2017•Int J BiologicalsRequires access
Isolation and purification of high purity thrombin
Hongbao Huang, Dengzhong Zheng, Xiulan Chen, Jing Zhao, Lili Zhang
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Abstract
Objective
To prepare high purity thrombin from porcine plasma.
Methods
Prothrombin was separated by DEAE-Sepharose FF anion-exchange chromatography from porcine plasma, and then was activated by CaCl2 to form thrombin. High purity thrombin was obtained by two-step SP-Sepharose FF cation-exchange chromatography from crude thrombin.
Results
The specific activity of thrombin after two-step SP-Sepharose FF cation-exchange chromatography was 2 546 IU/mg, 8 times of the crude thrombin activity. The recovery of thrombin was 54%. The result of fast protein liquid chromatography showed that the purity of thrombin after two-step SP-Sepharose FF cation-exchange chromatography was 89%.
Conclusion
High purity thrombin can be obtained by DEAE-Sepharose FF and SP-Sepharose FF ion-exchange chromatography from porcine plasma.
Key words:
Thrombin; Chromatography, ion exchange; Technology, pharmaceutical