Diagnostic Detection of Human Lung Cancer-Associated Antigen Using a Gold Nanoparticle-Based Electrochemical Immunosensor
Diagnostic Detection of Human Lung Cancer-Associated Antigen Using a Gold Nanoparticle-Based Electrochemical Immunosensor
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DOI：
10.1021/ac1001959
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发表时间：
2010-07-15
影响因子：
7.4
通讯作者：
Chou, Chien
中科院分区：
文献类型：
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作者：
Ho, Ja-an Annie;Chang, Heng-Chia;Chou, Chien
开发快速、灵敏的方法来检测免疫原性肿瘤相关抗原不仅对于理解它们在癌症免疫学中的作用很重要，而且对于临床诊断的发展也很重要。 α-烯醇化酶 (ENO1) 是一种 p48 分子，广泛分布于多种组织中，而 γ-烯醇化酶 (ENO2) 和 β-烯醇化酶 (ENO3) 分别仅存在于神经元/神经内分泌和肌肉组织中。由于ENO1与小细胞肺癌、非小细胞肺癌和头颈癌相关，因此它可以作为肺癌的潜在诊断标志物。在这项研究中，我们开发了一种简单、新颖且灵敏的电化学夹心免疫传感器，用于检测 ENO1；它通过抗ENO1单克隆抗体物理吸附在聚乙二醇修饰的一次性丝网印刷电极上作为检测平台，以多克隆二抗ENO1标记的金纳米粒子（AuNP）聚集作为电化学信号探针。聚集的 AuNP@抗体对样品 ENO1 的免疫识别发生在电极表面；将结合的 AuNP 聚集体在 1.2 V 的 0.1 M HCl 中氧化 120 秒，然后在方波伏安法 (SWV) 模式下还原 AuCl4- 后，获得来自结合的 AuNP 聚集体的电化学信号。由此产生的 S 形剂量响应曲线具有 10(-8) 至 10(-12) g/mL 的线性动态工作范围。这种基于 AuNP 聚集体的测定提供了一种用于检测痕量水平 ENO1 的扩增方法，导致检测限低至 11.9 fg（相当于 5 μL 2.38 pg/mL 溶液）。
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. a-Enolase (ENO1), a p48 molecule, is widely distributed in a variety of tissues, whereas gamma-enolase (ENO2) and beta-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 mu L of a 2.38 pg/mL solution).