亲和型生物传感器对TNF-α/sTNFR-I相互作用的研究  

Studied on interaction of an affinity biosensor against TNF-α/sTNFR-I

作　　者：李静[1] 吴宝艳[1] 黄加栋[1] 史海滨[1] 陈强[1] 

机构地区：[1]南开大学生命科学学院,天津300071

出　　处：《传感器与微系统》2006年第3期25-27,31,共4页Transducer and Microsystem Technologies

基　　金：国家自然科学基金资助项目(90209054)

摘　　要：应用亲和型生物传感器—Asys生物传感器实时监测不同浓度的肿瘤坏死因子-α(TNF-α)与可溶性肿瘤坏死因子受体-I(sTNFR-I)的亲和反应过程,并应用FASTfit软件对数据进行分析。结果表明:sTNFR-I以2.25 ng/mm2的密度固定于样品池敏感基片表面,TNF-α可与其发生特异性、浓度依赖性亲和反应,应用FASTfit软件进行数据拟合,并计算其反应的动力学常数,其反应符合二级动力学方程,结合速率常数kass=4.21×104L.mol-1.s-1,解离速率常数kd iss=2.12×10-2s-1,则解离平衡常数kD=5.05×10-7mol.L-1,结合平衡常数kA=1.99×106L.mol-1。应用IAsys传感器可实时监测结合和解离反应过程,操作简便、快速,是研究TNF-α和sTNFR相互作用动力学的有效方法。The interaction between different concentrations of tumor necrosis factor-α （TNF-α） and soluble tumors necrosis factor receptor-I （sTNFR-I） is studied by using an affinity biosensor-IAsys biosensor, and the data is analyzed by using the FASTfit program, sTNFR-I is immobilized on the cuvette surface with the density of 2.25 ng/mm^2 ,TNF-α then bound to immobilized sTNFR-I specifically in a dose-dependent manner. By FASTfit fitting,the responses fitted the biphasic association quite well, and the error i,s small and random. Kinetic constants for TNF-α binding to sTNFR-I are determined from the plots of kon versus＂ the concentration of TNF-α ： kmax =. 4.21 × 10^4L · mol^-1 · s^-1 ,kdisa =2.12 × 10^-2s^-1 ,kD =5.05 × 10^-7mol · L^-1 ,kA = 1.99 × 10^6L · mol^-1. iAsys biosensor is able to monitor the binding and dissociating courses in real time with simple and quick operation, which provided an effective assay for kinetic analysis of TNF-α/sTNFR-I interaction.

关 键 词：肿瘤坏死因子-Α 可溶性肿瘤坏死因子受体-I IAsys生物传感器 动力学分析 

分 类 号：TP212[自动化与计算机技术—检测技术与自动化装置]