Chinese Journal of Chromatography ›› 2013, Vol. 31 ›› Issue (8): 813-816.DOI: 10.3724/SP.J.1123.2013.01024

• Technical Notes • Previous Articles    

Preparation of thrombin affinity chromatography media

WANG Xiongfei, ZHANG Yujie, YE Jing, LUO Zhiqiang, ZHAN Xueyan, YUAN Ruijuan   

  1. School of Chinese Pharmacy, Beijing University of Chinese Medicine, Beijing 100102, China
  • Received:2013-01-11 Revised:2013-03-29 Online:2013-08-28 Published:2013-09-29
  • Contact: O658

Abstract:

The preparation of thrombin affinity chromatography media is described in this paper. First, the thrombin affinity chromatography media was prepared through thrombin coupled with cyanogen bromide activated agarose. And then the enzyme activity of thrombin on the coupled media was measured by chromogenic substrate method. The conditions were confirmed based on the enzyme activity of thrombin on the coupled media. The optimum conditions were as follows: the use of Na2CO3-NaHCO3 (containing 0.5 mol/L NaCl) solution of pH 8.3 as the reaction solution, 200 U thrombin per 1 g agarose, and reaction for 10 h at room temperature. The prepared affinity chromatography media had good stability, on which 70.6% of the enzyme activity of thrombin existed after 40 days’ storage. The thrombin affinity chromatography media can be widely applied in the screening and purification of thrombin inhibitors in natural products.

Key words: affinity chromatography, chromogenic substrate method, thrombin

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