高等学校化学学报 ›› 2002, Vol. 23 ›› Issue (3): 382.

• 研究简报 • 上一篇    下一篇

利用表面等离子体共振技术进行TNF免疫复合物的动力学分析

杨帆1, 崔小强1, 杨秀荣1, 陈婷2, 陆军2   

  1. 1. 中国科学院长春应用化学研究所电分析化学国家重点实验室, 国家电化学和光谱研究分析中心, 长春 130022;
    2. 东北师范大学遗传与细胞研究所, 长春 130024
  • 收稿日期:2000-12-20 出版日期:2002-03-24 发布日期:2002-03-24
  • 通讯作者: 杨秀荣(1946年生),女,博士,研究员,博士生导师,从事生物分析研究.
  • 基金资助:

    国家自然科学基金(批准号:20075027)资助

Kinetic Analysis of TNF Immune Complex by Using Surface Plasmon Resonance

YANG Fan1, CUI Xiao-Qiang1, YANG Xiu-Rong1, CHEN Ting2, LU Jun2   

  1. 1. State Key Lab of Electroanalytical Chemistry, National Analytical Research Center of Electrochemistryand Spectroscopy, Changchun Institute of App lied Chemistry, Chinese Academy of Sciences, Changchun 130022, China;
    2. Institute of Genetics and Cytology, Northeast Normal University, Changchun 130024, China
  • Received:2000-12-20 Online:2002-03-24 Published:2002-03-24

关键词: 表面等离子体共振, 肿瘤坏死因子, 动力学分析

Abstract: The kinetic analysis of the interaction between tumor necrosis factor(TNF) and its monoclonal antibody was performed by surface plasmon resonance(SPR) technique. The monoclonal antibody was immobilized to the surface of CM5 sensor chip by amine coupling. TNFat different concentrations was injected across the mAb immobilized surface. The interaction was recorded in real time and could be seen on the sensorgram. One cycle, including association, dissociation and regeneration, lasted no more than 15min . The interaction results was evaluated using 1:1 Langmuir binding model. The kinetic rate constants were calculated to be: ka=1.68×103 L·mol-1 ·s-1 , kd=1.73×10-4 s-1 , and the affinity constants KA=9.7×106 L·mol-1 , KD=1.03×10-7 mol·L-1 . The χ2 was 3.47, which showed that the interaction is consistent with the 1:1 model. We can see from the results that although there are two binding sites in one mAb molecule, TNFreacts with each site in an independent and noncooperative manner.

Key words: Surface plasmon resonance, Tumor necrosis factor, Kinetic analysis

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