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石英晶体微天平-电化学阻抗谱同步研究纳米材料修饰金电极表面抗人免疫球蛋白G的吸附及其免疫反应

Simultaneous quartz crystal microbalance-electrochemical impedance spectroscopy study on the adsorption of anti-human immunoglobulin G and its immunoreaction at nanomaterial-modified Au electrode surfaces.

作者信息

Jia Xueen, Xie Qingji, Zhang Youyu, Yao Shouzhuo

机构信息

Key Laboratory of Chemical Biology & Traditional Chinese Medicine Research (Ministry of Education), Hunan Normal University, P. R. China.

出版信息

Anal Sci. 2007 Jun;23(6):689-96. doi: 10.2116/analsci.23.689.

Abstract

The quartz crystal microbalance method (QCM), in combination with electrochemical impedance spectroscopy (EIS), has been utilized to monitor in situ anti-human IgG adsorption on several Au-based surfaces, bare Au, nanogold/4-aminothiophenol (4AT)/Au, and multi-walled carbon nanotubes (MWCNT)/Au, and succeeding human IgG reactions. Also, the immobilization protocol of anti-human IgG via its glutaraldehyde (GA) cross-linking with self-assembled 4AT on an Au electrode and the subsequent surface immunoreaction were examined. The resonant frequency (f(0)) and the motional resistance (R(1)) of the piezoelectric quartz crystal (PQC) as well as electrochemical impedance parameters were measured and discussed. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) of the ferricyanide/ferrocyanide couple were examined before and after electrode modification, the antibody adsorption and antibody-antigen reactions. We found that the amount for antibody adsorption was the greatest on the colloid Au modified surface, and that at MWCNT ranked the second, while specific bioactivity was almost identical on the four kinds of surfaces. Two parameters simultaneously obtained at the colloid Au modified surface, Deltaf(0) and DeltaC(s) (interfacial capacitance), have been used to estimate the association constant of the immunoreaction.

摘要

石英晶体微天平法(QCM)与电化学阻抗谱(EIS)相结合,已被用于原位监测抗人IgG在几种金基表面(裸金、纳米金/4-氨基硫酚(4AT)/金和多壁碳纳米管(MWCNT)/金)上的吸附情况以及后续的人IgG反应。此外,还研究了通过戊二醛(GA)将抗人IgG与自组装在金电极上的4AT交联的固定化方案以及随后的表面免疫反应。测量并讨论了压电石英晶体(PQC)的共振频率(f(0))和运动电阻(R(1))以及电化学阻抗参数。在电极修饰、抗体吸附和抗体-抗原反应之前和之后,检测了铁氰化物/亚铁氰化物电对的循环伏安法(CV)和电化学阻抗谱(EIS)。我们发现,抗体在胶体金修饰表面上的吸附量最大,在MWCNT上的吸附量次之,而四种表面上的特异性生物活性几乎相同。在胶体金修饰表面同时获得的两个参数,即Δf(0)和ΔC(s)(界面电容),已被用于估计免疫反应的缔合常数。

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