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  

Kinetic Analysis of TNF Immune Complex by Using Surface Plasmon Resonance

作　　者：杨帆[1] 崔小强[1] 杨秀荣[1] 陈婷[2] 陆军[2] 

机构地区：[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

出　　处：《高等学校化学学报》2002年第3期382-384,共3页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金 (批准号 :2 0 0 75 0 2 7)资助

摘　　要：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. TNF at 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 15 min . The interaction results was evaluated using 1∶1 Langmuir binding model. The kinetic rate constants were calculated to be: k a=1.68×10 3 L·mol -1 ·s -1 , k d=1.73×10 -4 s -1 , and the affinity constants K A=9.7×10 6 L·mol -1 , K D=1.03×10 -7 mol·L -1 . The χ \+2 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, TNF reacts with each site in an independent and noncooperative manner.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. TNF at 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 15 min . The interaction results was evaluated using 1∶1 Langmuir binding model. The kinetic rate constants were calculated to be: k a=1.68×10 3 L·mol -1 ·s -1 , k d=1.73×10 -4 s -1 , and the affinity constants K A=9.7×10 6 L·mol -1 , K D=1.03×10 -7 mol·L -1 . The χ \+2 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, TNF reacts with each site in an independent and noncooperative manner.

关 键 词：表面等离子体共振 肿瘤坏死因子 动力学分析 TNF免疫复合物 亲合力 分子间相互作用 抗肿瘤药物 生物传感器 

分 类 号：R979.1[医药卫生—药品]