Citation: WANG Xiongfei, ZHANG Yujie, YE Jing, LUO Zhiqiang, ZHAN Xueyan, YUAN Ruijuan. Preparation of thrombin affinity chromatography media[J]. Chinese Journal of Chromatography, ;2013, 31(8): 813-816. doi: 10.3724/SP.J.1123.2013.01024 shu

Preparation of thrombin affinity chromatography media

1.
School of Chinese Pharmacy, Beijing University of Chinese Medicine, Beijing 100102, China

Corresponding author: YUAN Ruijuan,
Received Date: 11 January 2013
Available Online: 29 March 2013
Fund Project: 北京市自然科学基金项目(7133247) (7133247)北京中医药大学自主选题项目(2011-JSBZZ-JS047) (2011-JSBZZ-JS047)国家级大学生创新创业训练项目(201210026034). (201210026034)

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.
- thrombin,
- affinity chromatography,
- chromogenic substrate method
1. [1]
  [1] Campbell D H, Luescher E, Lerman L S. Proc Natl Acad Sci USA, 1951, 37(9): 575
2. [2]
  [2] Prasanna R R, Vijayalakshmi M A. Pure Appl Chem, 2010, 82(1): 39
3. [3]
  [3] Cheung R C F, Wong J H, Ng T B. Appl Microbiol Biot, 2012, 96(6): 1411
4. [4]
  [4] Leitner A. TrAC-Trend Anal Chem, 2010, 29(2): 177
5. [5]
  [5] Marchetti N, Caciolli L, Lagana A, et al. Anal Chem, 2012, 84(16): 7138
6. [6]
  [6] Yang H, Gurgel P V, Carbonell R G. J Chromatogr A, 2009, 1216(6): 910
7. [7]
  [7] Hage D S, Anguizola J A, Bi C, et al. J Pharmaceut Biomed, 2012, 69: 93
8. [8]
  [8] Chen Y H, Huang H M, Li R Q. Chinese Journal of Chromatography (陈义烘, 黄慧敏, 李任强. 色谱), 2007, 25(3): 332
9. [9]
  [9] Mao S J T, Yates M T, Owen T J, et al. Biochemistry, 1988, 27: 8170
10. [10]
  [10] Electricwala A, Sawyer R T, Jones C P, et al. Blood Coagul Fibrin, 1991, 2(1): 83
11. [11]
  [11] Tong X R, Wang D B, Ma S B. Journal of Kunming Teachers College (仝向荣, 王德斌, 马绍宾. 昆明师范高等专科学校学报), 2004, 26(4): 70
12. [12]
  [12] Strube K H, Kroger B, Bialojan S, et al. J Biol Chem, 1993, 268(12): 8590
13. [13]
  [13] Rogero C, Chaffey B T, Mateo-Marti E, et al. Phys Chem C, 2008, 112(25): 9308
14. [14]
  [14] Hage D S. Encyclopedia of Analytical Chemistry: Affinity Chromatography. New York: John Wiley & Sons, 2006
15. [15]
  [15] Deng Z Q, Qu X J, Ouyang Y et al. Acta Academiae Medicinae Hubei (邓兆群, 屈雪菊, 欧阳怡, 等. 湖北医科大学学报), 1996, 17(2): 195
1. [1]
  Qiuting Zhang , Fan Wu , Jin Liu , Zian Lin . Chromatographic Stationary Phase and Chiral Separation Using Frame Materials. University Chemistry, 2025, 40(4): 291-298. doi: 10.12461/PKU.DXHX202405174
2. [2]
  Yangrui Xu , Yewei Ren , Xinlin Liu , Hongping Li , Ziyang Lu . NH₂-UIO-66 Based Hydrophobic Porous Liquid with High Mass Transfer and Affinity Surface for Enhancing CO₂ Photoreduction. Acta Physico-Chimica Sinica, 2024, 40(11): 2403032-0. doi: 10.3866/PKU.WHXB202403032
3. [3]
  Houjin Li , Lin Wu , Xingwen Sun , Yuan Zheng , Zhanxiang Liu , Shuanglian Cai , Ying Xiong , Guangao Yu , Qingwen Liu , Jie Han , Xin Du , Chengshan Yuan , Qihan Zhang , Jianrong Zhang , Shuyong Zhang . Basic Operations and Specification Suggestions for Organic Chemical Chromatography Experiments. University Chemistry, 2025, 40(5): 93-105. doi: 10.12461/PKU.DXHX202408100
4. [4]
  Runjie Li , Hang Liu , Xisheng Wang , Wanqun Zhang , Wanqun Hu , Kaiping Yang , Qiang Zhou , Si Liu , Pingping Zhu , Wei Shao . 氨基酸的衍生及手性气相色谱分离创新实验. University Chemistry, 2025, 40(6): 286-295. doi: 10.12461/PKU.DXHX202407059
5. [5]
  .
  CCS Chemistry | 超分子活化底物为自由基促进高效选择性光催化氧化
  . CCS Chemistry, 2025, 7(10.31635/ccschem.025.202405229): -.
6. [6]
  Jingming Li , Bowen Ding , Nan Li , Nurgul . Application of Comparative Teaching Method in Experimental Project Design of Instrumental Analysis Course: A Case Study in Chromatography Experiment Teaching. University Chemistry, 2024, 39(8): 263-269. doi: 10.3866/PKU.DXHX202312078
7. [7]
  Quanliang Chen , Zhaohui Zhou . Research on the Active Site of Nitrogenase over Fifty Years. University Chemistry, 2024, 39(7): 287-293. doi: 10.3866/PKU.DXHX202310133
8. [8]
  Zunxiang Zeng , Yuling Hu , Yufei Hu , Hua Xiao . Analysis of Plant Essential Oils by Supercritical CO₂Extraction with Gas Chromatography-Mass Spectrometry: An Instrumental Analysis Comprehensive Experiment Teaching Reform. University Chemistry, 2024, 39(3): 274-282. doi: 10.3866/PKU.DXHX202309069
9. [9]
  Yanhui Zhong , Ran Wang , Zian Lin . Analysis of Halogenated Quinone Compounds in Environmental Water by Dispersive Solid-Phase Extraction with Liquid Chromatography-Triple Quadrupole Mass Spectrometry. University Chemistry, 2024, 39(11): 296-303. doi: 10.12461/PKU.DXHX202402017
10. [10]
  Fan Wu , Wenchang Tian , Jin Liu , Qiuting Zhang , YanHui Zhong , Zian Lin . Core-Shell Structured Covalent Organic Framework-Coated Silica Microspheres as Mixed-Mode Stationary Phase for High Performance Liquid Chromatography. University Chemistry, 2024, 39(11): 319-326. doi: 10.12461/PKU.DXHX202403031
11. [11]
  Shunü Peng , Huamin Li , Zhaobin Chen , Yiru Wang . Simultaneous Application of Multiple Quantitative Analysis Methods in Gas Chromatography for the Determination of Active Ingredients in Traditional Chinese Medicine Preparations. University Chemistry, 2025, 40(10): 243-249. doi: 10.12461/PKU.DXHX202412043
12. [12]
  Yifan Xie , Liyun Yao , Ruolin Yang , Yuxing Cai , Yujie Jin , Ning Li . Exploration and Practice of Online and Offline Hybrid Teaching Mode in High-Performance Liquid Chromatography Experiment. University Chemistry, 2025, 40(11): 100-107. doi: 10.12461/PKU.DXHX202412133
13. [13]
  Zhou Fang , Zhihao Zhang , Weihan Jiang , Kin Shing Chan . Warfarin: From Poison to Cure, the Remarkable Journey of a Molecule. University Chemistry, 2025, 40(4): 326-330. doi: 10.12461/PKU.DXHX202406038
14. [14]
  Heng Zhang . Determination of All Rate Constants in the Enzyme Catalyzed Reactions Based on Michaelis-Menten Mechanism. University Chemistry, 2024, 39(4): 395-400. doi: 10.3866/PKU.DXHX202310047
15. [15]
  Yerong Chen , Bingbin Yang , Xinglei He , Yuqi Lin , Keyin Ye . Enzyme-Directed Evolution Enables Bioconversion of Organosilicon Compounds. University Chemistry, 2025, 40(10): 121-129. doi: 10.12461/PKU.DXHX202411054
16. [16]
  Chunmei GUO , Weihan YIN , Jingyi SHI , Jianhang ZHAO , Ying CHEN , Quli FAN . Facile construction and peroxidase-like activity of single-atom platinum nanozyme. Chinese Journal of Inorganic Chemistry, 2024, 40(9): 1633-1639. doi: 10.11862/CJIC.20240162
17. [17]
  Yu Dai , Xueting Sun , Haoyu Wu , Naizhu Li , Guoe Cheng , Xiaojin Zhang , Fan Xia . Determination of the Michaelis Constant for Gold Nanozyme-Catalyzed Decomposition of Hydrogen Peroxide. University Chemistry, 2025, 40(5): 351-356. doi: 10.12461/PKU.DXHX202407052
18. [18]
  Pengli GUAN , Renhu BAI , Xiuling SUN , Bin LIU . Trianiline-derived aggregation-induced emission luminogen probe for lipase detection and cell imaging. Chinese Journal of Inorganic Chemistry, 2025, 41(9): 1817-1826. doi: 10.11862/CJIC.20250058
19. [19]
  Hui Shi , Shuangyan Huan , Yuzhi Wang . Ideological and Political Design of Potassium Permanganate Oxidation-Reduction Titration Experiment. University Chemistry, 2024, 39(2): 175-180. doi: 10.3866/PKU.DXHX202308042
20. [20]
  Zhenlin Zhou , Siyuan Chen , Yi Liu , Chengguo Hu , Faqiong Zhao . A New Program of Voltammetry Experiment Teaching Based on Laser-Scribed Graphene Electrode. University Chemistry, 2024, 39(2): 358-370. doi: 10.3866/PKU.DXHX202308049

Get Citation

PDF

XML

Metrics

PDF Downloads(0)
Abstract views(371)
HTML views(28)

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142