多壁碳纳米管修饰铂基片石英晶体微天平研究白细胞介素-6与其受体的相互作用

摘要

An affinity biosensor using multi-walled carbon nanotubes to activate Pt quartz crystal microbalance (QCM) electrode and immobilizing interleukin-6 (IL-6) on 1-ethyl-3-(3-dimethylamin~ opropyl) carbodimide (EDC)/N-hydroxysuccinimide (NHS)-activated QCM electrode was employed to real time detect the interaction and kinetic analysis of IL-6 and soluble interleukin-6 receptor (sIL-6R), and the association data between IL-6 and sIL-6R was fitted by Origin software. The results showed that the combined masses of sIL-6R on IL-6-EDC/NHS-MWCNTs activated Pt QCM increased linearly with the increase of concentration of sIL-6R, and the linear correlation coefficient was 0. 997. For the interaction, KA was 3. 37 × 106 L/mol, and KD was 2. 97 × 10-~7 mol/L. The experiment proves the feasibility of using multi-walled carbon nanotubes activated Pt quartz crystal microbalance to monitor the interaction and kinetic analysis of IL-6 and sIL-6R, it can help to clarify the important biological function of IL-6 and sIL-6R. All experiments were performed in PBS (0. 01 mol/L, pH 7. 4) at room temperature.%在pH 7.4、室温条件下,用多壁碳纳米管(MWCNTs)修饰石英晶体微天平(QCM) Pt基片,经过EDC/NHS活化后固定白细胞介素-6(IL-6),检测不同浓度可溶性白细胞介素-6受体(sIL-6R)与IL-6的结合所引起响应信号的变化,并用Origin软件对数据进行拟合分析,构建了一种高性能实时监测IL-6与sIL-6R之间相互作用的亲和型生物传感器.实验表明:随着sIL-6R浓度的增加,其结合量呈线性增长,相关系数(R2)为0.997.IL-6与sIL-6R之间相互作用的结合平衡常数(KA)和解离平衡常数(KD)分别为3.37×106 L/mol和2.97×10-7 mol/L.验证了通过MWCNTs修饰的Pt QCM能够高效监测IL-6与sIL-6R间相互作用及动力学过程,有助于阐明二者的生物学功能.