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一种基于FeO@Au的伪均相电化学免疫传感器,用于以甲胎蛋白抗体-金纳米颗粒-辣根过氧化物酶作为检测探针测定甲胎蛋白。

A FeO@Au-basedpseudo-homogeneous electrochemical immunosensor for AFP measurement using AFP antibody-GNPs-HRP as detection probe.

作者信息

Yuan Yulin, Li Shanshan, Xue Yewei, Liang Jintao, Cui Lijie, Li Qingbo, Zhou Sufang, Huang Yong, Li Guiyin, Zhao Yongxiang

机构信息

Department of Clinical Laboratory, The People's Hospital of Guangxi Zhuang Autonomous Region, Nanning, Guangxi 530021, China.

School of Life and Environmental Sciences, Guilin University of Electronic Technology, Guilin, Guangxi 541004, China.

出版信息

Anal Biochem. 2017 Oct 1;534:56-63. doi: 10.1016/j.ab.2017.07.015. Epub 2017 Jul 13.

Abstract

In this study, a FeO@Au-based pseudo-homogeneous electrochemical immunosensor was prepared for detection of alpha fetoprotein (AFP), a well-known hepatocellular carcinoma biomarker. The primary antibody (Ab1) was immobilized on FeO@Au NPs as the capture probe. Horseradish peroxidase (HRP) and secondary antibody (Ab2) were conjugated on gold nanoparticles (GNPs) through electrostatic adsorption to form signal-amplifying labels. In the presence of AFP, a sandwich immunocomplex was formed via specific recognition of antigen-antibody in a FeO@Au-basedpseudo-homogeneousreaction system. After the immunocomplex was captured to the surface of magnetic glassy carbon electrode (MGCE), the labeling HRP catalyzed the decomposition of HO, resulting in a substantial current for the quantitative detection of AFP. The amperometric (i-t) method was employed to record the response signal of the immunosensor based on the catalysis of the immobilized HRP toward the reduction of HO with hydroquinone (HQ) as the redox mediator. Under the optimal conditions, the amperometric current response presented a linear relationship with AFP concentration over the range of 20 ng/mL-100 ng/mLwith a correlation coefficient of 0.9940, and the detection limit was 0.64 ng/mL at signal/noise [S/N] = 3. Moreover, the electrochemical immunosensor exhibited higher anti-interference ability, acceptable reproducibility and long-term stability for AFP detection.

摘要

在本研究中,制备了一种基于FeO@Au的准均相电化学免疫传感器,用于检测甲胎蛋白(AFP),这是一种著名的肝细胞癌生物标志物。将一抗(Ab1)固定在FeO@Au纳米颗粒上作为捕获探针。辣根过氧化物酶(HRP)和二抗(Ab2)通过静电吸附结合在金纳米颗粒(GNP)上,形成信号放大标记。在AFP存在的情况下,通过基于FeO@Au的准均相反应体系中抗原-抗体的特异性识别形成夹心免疫复合物。将免疫复合物捕获到磁性玻碳电极(MGCE)表面后,标记的HRP催化H₂O₂分解,产生大量电流用于AFP的定量检测。采用安培(i-t)法,以对苯二酚(HQ)作为氧化还原介质,基于固定化HRP对H₂O₂还原的催化作用记录免疫传感器的响应信号。在最佳条件下,安培电流响应与AFP浓度在20 ng/mL - 100 ng/mL范围内呈线性关系,相关系数为0.9940,在信噪比[S/N]=3时检测限为0.64 ng/mL。此外,该电化学免疫传感器对AFP检测表现出更高的抗干扰能力、可接受的重现性和长期稳定性。

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