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基于金纳米粒子掺入壳聚糖生物聚合物纳米复合材料膜的电化学免疫传感器的研制及其在以 PSA 为生物标志物的前列腺癌检测中的应用。

Development of an electrochemical immunosensor based on gold nanoparticles incorporated chitosan biopolymer nanocomposite film for the detection of prostate cancer using PSA as biomarker.

机构信息

Electroanalytical Lab, Department of Chemistry, K L University, Guntur, Andhra Pradesh, India; Department of Biotechnology, K L University, Guntur, Andhra Pradesh, India.

Electroanalytical Lab, Department of Chemistry, K L University, Guntur, Andhra Pradesh, India.

出版信息

Enzyme Microb Technol. 2018 May;112:43-51. doi: 10.1016/j.enzmictec.2017.10.009. Epub 2017 Oct 24.

Abstract

In this study, we have developed an electrochemical immunosensor for the detection of prostate-specific antigen (PSA) based on gold nanoparticles (AuNPs) and a chitosan (CHI) nanocomposite film coated on a screen printed electrode (SPE). The modified SPE was characterized using scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The immunosensor was fabricated by sequential immobilization of anti-PSA primary antibody, PSA antigen and HRP-tagged secondary antibody on the surface of AuNPs/CHI/SPE. When this immunosensor was placed in 2.5mM HO, as HRP substrate, and 1mM methylene blue (MB), as redox mediator, greatly amplified immunosensing signals were observed. Immunosensing signals for the reduction of HO were monitored in phosphate buffer (pH 7.0) by CV and square wave voltammetry (SWV). Under optimized conditions steady state current increased linearly with PSA concentration over the range 1-18ng/ml with a detection limit of 0.001ng/ml. The fabricated immunosensor demonstrated excellent sensitivity, stability and reproducibility and therefore was successfully applied to analyse PSA in biological samples. Finally, the results indicate that the proposed immunosensor has potential in clinical screening of cancer biomarkers.

摘要

在这项研究中,我们开发了一种基于金纳米粒子(AuNPs)和壳聚糖(CHI)纳米复合材料膜的电化学免疫传感器,用于检测前列腺特异性抗原(PSA)。修饰后的电极采用扫描电子显微镜(SEM)、能谱(EDX)、循环伏安法(CV)和电化学阻抗谱(EIS)进行了表征。通过将抗 PSA 一抗、PSA 抗原和 HRP 标记的二抗依次固定在 AuNPs/CHI/SPE 表面,制备了免疫传感器。当将该免疫传感器置于 2.5mM HO 作为 HRP 底物和 1mM 亚甲基蓝(MB)作为氧化还原介体时,观察到了大大放大的免疫传感信号。通过 CV 和方波伏安法(SWV)在磷酸盐缓冲液(pH 7.0)中监测 HO 的还原免疫传感信号。在优化条件下,稳态电流在 1-18ng/ml 范围内与 PSA 浓度呈线性关系,检测限为 0.001ng/ml。所制备的免疫传感器表现出优异的灵敏度、稳定性和重现性,因此成功应用于生物样品中 PSA 的分析。最后,结果表明,所提出的免疫传感器在癌症生物标志物的临床筛选中具有潜力。

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