【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。更多还原

【Abstract】 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/mm²）,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 is small and random.Kinetic constants for TNF-α binding to sTNFR-I are determined from the plots of k_on versus the concentration of TNF-α:k_ass=（4.21×10⁴L）·mol^-1·s^-1,k_diss=2.12×10^-2s^-1,k_D=5.05×10^-7mol·L^-1,k_A=1.99×10⁶L·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.更多还原