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基于 ITO 电极修饰的还原氧化石墨烯-金纳米复合物和 CdS 标记的二抗用于阿尔茨海默病生物标志物 S100β蛋白的光电化学三明治免疫分析

A photoelectrochemical sandwich immunoassay for protein S100β, a biomarker for Alzheimer's disease, using an ITO electrode modified with a reduced graphene oxide-gold conjugate and CdS-labeled secondary antibody.

机构信息

Departamento de Ingeniería Electrónica, Eléctrica y Automática, Universitat Rovira i Virgili, Avda. Països Catalans 26, 43007, Tarragona, Spain.

出版信息

Mikrochim Acta. 2019 Jan 16;186(2):117. doi: 10.1007/s00604-018-3159-x.

Abstract

A sandwich-type photoelectrochemical immunoassay is described for the protein S100ß which is an Alzheimer's disease biomarker found in the astrocytes of the brain. Antibody against S100ß (anti-S100ß) was labeled with CdS quantum dots and then acted as a secondary antibody. The labeled antibody was characterized by FTIR, ultraviolet-visible and fluorescence spectroscopy. An indium-tin oxide (ITO) electrode was modified with a nanocomposite prepared from reduced graphene oxide and gold nanoparticles. Then, a sol-gel film containing isocyanate functional groups (-N=C=O) was cast on the surface of the electrode. The NCO group reacts with amino groups of the labeled antibody to covalently bind them to the surface. The S100β was bound by the primary immobilized antibody on the rGO-Au/ITO electrode and then sandwiched with the labeled secondary antibody. Cyclic voltammetry and electrochemical impedance spectroscopy were applied to confirm the stepwise changes in the electrochemical properties of the electrode surface. The photoelectrochemical immunoassay, typically operated at a potential of +0.2 V (vs. Ag|AgCl) gives a signal that is related to the logarithm of the S100β concentration in the range from 0.25 to 10 ng·mL with a lower detection limit of 0.15 pg·mL. The method was successfully applied to the determination of S100β in human serum samples. Graphical abstract Schematic presentation of an immunosensor which is based on an indium tin oxide modified with reduced graphene oxide decorated with gold nanocomposite and antibody. The immunosensor was applied for the determination of S100β biomarker by using in the labeled antibody.

摘要

夹心型光电化学免疫分析用于检测 S100ß,S100ß 是大脑星形胶质细胞中发现的阿尔茨海默病生物标志物。S100ß 的抗体(抗 S100ß)被硫化镉量子点标记,然后作为二次抗体。通过傅里叶变换红外光谱、紫外-可见光谱和荧光光谱对标记抗体进行了表征。将氧化铟锡(ITO)电极用还原氧化石墨烯和金纳米粒子的纳米复合材料进行修饰。然后,在电极表面涂覆含有异氰酸酯官能团(-N=C=O)的溶胶-凝胶膜。NCO 基团与标记抗体的氨基反应,将其共价结合到表面上。S100β 与固定在 rGO-Au/ITO 电极上的初级抗体结合,然后与标记的次级抗体夹在一起。循环伏安法和电化学阻抗谱用于证实电极表面电化学性质的逐步变化。光电化学免疫分析通常在+0.2 V(相对于 Ag|AgCl)的电位下操作,其信号与 S100β 浓度的对数在 0.25 至 10 ng·mL 范围内相关,检测限低至 0.15 pg·mL。该方法成功应用于人血清样品中 S100β 的测定。

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