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]
  Yangrui Xu , Yewei Ren , Xinlin Liu , Hongping Li , Ziyang Lu . 具有高传质和亲和表面的NH₂-UIO-66基疏水多孔液体用于增强CO₂光还原. Acta Physico-Chimica Sinica, 2024, 40(11): 2403032-. doi: 10.3866/PKU.WHXB202403032
2. [2]
  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
3. [3]
  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
4. [4]
  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
5. [5]
  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
6. [6]
  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
7. [7]
  .
  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): -.
8. [8]
  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
9. [9]
  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
10. [10]
  Liwei Wang , Guangran Ma , Li Wang , Fugang Xu . A Comprehensive Analytical Chemistry Experiment: Colorimetric Detection of Vitamin C Using Nanozyme and Smartphone. University Chemistry, 2024, 39(8): 255-262. doi: 10.3866/PKU.DXHX202312094
11. [11]
  Zhuoya WANG , Le HE , Zhiquan LIN , Yingxi WANG , Ling LI . Multifunctional nanozyme Prussian blue modified copper peroxide: Synthesis and photothermal enhanced catalytic therapy of self-provided hydrogen peroxide. Chinese Journal of Inorganic Chemistry, 2024, 40(12): 2445-2454. doi: 10.11862/CJIC.20240194
12. [12]
  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
13. [13]
  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
14. [14]
  Feng Liang , Desheng Li , Yuting Jiang , Jiaxin Dong , Dongcheng Liu , Xingcan Shen . Method Exploration and Instrument Innovation for the Experiment of Colloid ζ Potential Measurement by Electrophoresis. University Chemistry, 2024, 39(5): 345-353. doi: 10.3866/PKU.DXHX202312009
15. [15]
  Wei Peng , Baoying Wen , Huamin Li , Yiru Wang , Jianfeng Li . Exploration and Practice on Raman Scattering Spectroscopy Experimental Teaching. University Chemistry, 2024, 39(8): 230-240. doi: 10.3866/PKU.DXHX202312062
16. [16]
  Yujia Luo , Yunpeng Qi , Huiping Xing , Yuhu Li . The Use of Viscosity Method for Predicting the Life Expectancy of Xuan Paper-based Heritage Objects. University Chemistry, 2024, 39(8): 290-294. doi: 10.3866/PKU.DXHX202401037
17. [17]
  Tiejun Su . The Construction and Application of the Calculation Formula for Endpoint Error in Precipitation Titration: A Case Study of the Mohr Method. University Chemistry, 2024, 39(11): 384-387. doi: 10.12461/PKU.DXHX202402039
18. [18]
  Jiahao Lu , Xin Ming , Yingjun Liu , Yuanyuan Hao , Peijuan Zhang , Songhan Shi , Yi Mao , Yue Yu , Shengying Cai , Zhen Xu , Chao Gao . 基于稳态电热法的石墨烯膜导热系数的精确可靠测量. Acta Physico-Chimica Sinica, 2025, 41(5): 100045-. doi: 10.1016/j.actphy.2025.100045
19. [19]
  Bingliang Li , Yuying Han , Dianyang Li , Dandan Liu , Wenbin Shang . One-Step Synthesis of Benorilate Guided by Green Chemistry Principles and in vivo Dynamic Evaluation. University Chemistry, 2024, 39(6): 342-349. doi: 10.3866/PKU.DXHX202311070
20. [20]
  Mengyao Shi , Kangle Su , Qingming Lu , Bin Zhang , Xiaowen Xu . Determination of Potassium Content in Tobacco Stem Ash by Flame Atomic Absorption Spectroscopy. University Chemistry, 2024, 39(10): 255-260. doi: 10.12461/PKU.DXHX202404105

Get Citation

PDF

XML

Metrics

PDF Downloads(0)
Abstract views(166)
HTML views(18)

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