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金纳米粒子作为电化学标签,用于在一次性芯片上进行固有互扰-free 的多重免疫分析。

Gold nanoparticle as an electrochemical label for inherently crosstalk-free multiplexed immunoassay on a disposable chip.

机构信息

Key Laboratory of Analytical Chemistry for Life Science (Ministry of Education of China), Department of Chemistry, Nanjing University, Hankou Road 22, Nanjing 210093, PR China.

出版信息

Anal Chim Acta. 2010 May 7;666(1-2):97-101. doi: 10.1016/j.aca.2010.03.060. Epub 2010 Apr 4.

Abstract

This work proposed a simple, sensitive and low-cost multiplexed immunoassay by combining a disposable chip with gold nanoparticle (AuNP) as an electrochemical label. The immunosensors array as the disposable chip was firstly prepared by immobilizing capture antibodies on different screen printed carbon working electrodes by passive adsorption. With a sandwich mode, the analytes were then bound to the corresponding capture antibodies for further capture of the gold nanoparticle labeled antibodies. Gold nanoparticles were finally electrooxidized in 0.1 M HCl to produce AuCl(4)(-) for differential pulse voltammetric detection. Using human IgG and goat IgG as model targets, under optimal conditions this method achieved linear ranges from 5.0 to 500 and 5.0 to 400 ng mL(-1) with limits of detection of 1.1 and 1.6 ng mL(-1), respectively. This method eliminated completely electrochemical crosstalk between adjacent immunosensors due to the strong adsorption of the AuCl(4)(-) on the printed carbon surfaces. The immunosensors array showed acceptable precision, reproducibility and stability, and could be readily applied to multianalyte determination for clinical diagnostics.

摘要

本工作提出了一种简单、灵敏且低成本的多重免疫分析方法,该方法将一次性芯片与金纳米粒子(AuNP)结合作为电化学标记物。将免疫传感器阵列作为一次性芯片,通过将捕获抗体通过被动吸附固定在不同的丝网印刷碳工作电极上进行制备。采用三明治模式,使分析物与相应的捕获抗体结合,进一步捕获金纳米粒子标记的抗体。最后,金纳米粒子在 0.1 M HCl 中电氧化生成 AuCl(4)(-)进行差分脉冲伏安法检测。用人 IgG 和羊 IgG 作为模型靶标,在最佳条件下,该方法的线性范围分别为 5.0 至 500 ng mL(-1)和 5.0 至 400 ng mL(-1),检测限分别为 1.1 和 1.6 ng mL(-1)。由于 AuCl(4)(-)在印刷碳表面的强吸附作用,该方法完全消除了相邻免疫传感器之间的电化学串扰。免疫传感器阵列表现出良好的精密度、重现性和稳定性,可方便地应用于临床诊断的多分析物测定。

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