Diagnostic Detection of Human Lung Cancer-Associated Antigen Using a Gold Nanoparticle-Based Electrochemical Immunosensor

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• 作者：Ja-an Annie Ho ; Heng-Chia Chang ; Neng-Yao Shih ; Li-Chen Wu ; Ying-Feng Chang ; Chii-Chang Chen ; Chien Chou
• 刊名：Analytical Chemistry
• 出版年：2010
• 出版时间：July 15, 2010
• 年：2010
• 卷：82
• 期：14
• 页码：5944-5950
• 全文大小：247K
• 年卷期：v.82,no.14(July 15, 2010)
• ISSN：1520-6882

文摘

The development of rapid and sensitive methods for the detection of immunogenic tumor-associated antigen is important not only for understanding their roles in cancer immunology but also for the development of clinical diagnostics. α-Enolase (ENO1), a p48 molecule, is widely distributed in a variety of tissues, whereas γ-enolase (ENO2) and β-enolase (ENO3) are found exclusively in neuron/neuroendocrine and muscle tissues, respectively. Because ENO1 has been correlated with small cell lung cancer, nonsmall cell lung cancer, and head and neck cancer, it can be used as a potential diagnostic marker for lung cancer. In this study, we developed a simple, yet novel and sensitive, electrochemical sandwich immunosensor for the detection of ENO1; it operates through physisorption of anti-ENO1 monoclonal antibody on polyethylene glycol-modified disposable screen-printed electrode as the detection platform, with polyclonal secondary anti-ENO1-tagged, gold nanoparticle (AuNP) congregates as electrochemical signal probes. The immunorecognition of the sample ENO1 by the congregated AuNP@antibody occurred on the surface of the electrodes; the electrochemical signal from the bound AuNP congregates was obtained after oxidizing them in 0.1 M HCl at 1.2 V for 120 s, followed by the reduction of AuCl⁴⁻ in square wave voltammetry (SWV) mode. The resulting sigmoidally shaped dose−response curves possessed a linear dynamic working range from 10⁻⁸ to 10⁻¹² g/mL. This AuNP congregate-based assay provides an amplification approach for detecting ENO1 at trace levels, leading to a detection limit as low as 11.9 fg (equivalent to 5 μL of a 2.38 pg/mL solution).