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用于检测帕金森病生物标志物的纳米结构无标记电化学免疫传感器。

Nanostructured label-free electrochemical immunosensor for detection of a Parkinson's disease biomarker.

机构信息

LEPABE, Department of Chemical Engineering, Faculty of Engineering of the University of Porto, Rua Dr. Roberto Frias, 4200-465, Porto, Portugal; REQUIMTE-LAQV, School of Engineering, Polytechnic Institute of Porto, R. Dr. António Bernardino de Almeida 431, 4200-072, Porto, Portugal.

LEPABE, Department of Chemical Engineering, Faculty of Engineering of the University of Porto, Rua Dr. Roberto Frias, 4200-465, Porto, Portugal.

出版信息

Talanta. 2023 Jan 15;252:123838. doi: 10.1016/j.talanta.2022.123838. Epub 2022 Aug 13.

Abstract

Aggregation of α-synuclein has been recognized as a critical event in the pathogenesis of Parkinson's disease whose prevalence is increasing with great socio-economic challenges for future generations. Here, we developed a sensitive and specific electrochemical immunosensor for the detection and quantification of this biomarker, based on the voltammetric study of a redox indicator signal, which decreases upon the analyte recognition by the antibody due to the electronic resistance increase. The proposed immunosensor is based on a screen-printed carbon electrode modified in a layer-by-layer approach, which through extensive characterization led to the successful nanostructuration of the transducer, through the drop-cast of 3.0 μL of a 0.1 mg mL single-walled carbon nanotubes suspension followed by electrodeposition of gold nanoparticles in a 3 mM HAuCl4 solution under a -0.2 V potential for 150 s. Monoclonal antibodies were immobilized on the gold nanoparticles surface through chemical modification at an optimal concentration of 200 μg mL. Using the proposed immunosensor, α-synuclein was detected in the range of 0.01-10 ng mL with a 4.1 and 12.6 pg mL limits of detection and quantification, respectively. Recovery values of 96.7, 106.2 and 102.9% were attained for the tested concentrations spiked in fetal bovine serum while also presenting excellent specificity and stability throughout one month. The nanostructured immunosensor provided a great interface for electronic transduction and biological recognition events, which enabled fast, sensitive and specific detection of α-synuclein while being based on a simple and inexpensive technology requiring small sample volumes, crucial characteristics for application in point-of-care testing.

摘要

α- 突触核蛋白的聚集已被认为是帕金森病发病机制中的一个关键事件,其患病率随着未来几代人的巨大社会经济挑战而增加。在这里,我们开发了一种基于电化学免疫传感器的敏感和特异性检测方法,用于检测和定量这种生物标志物,该传感器基于电化学指示剂信号的伏安研究,由于抗体与分析物的相互作用导致电子电阻增加,该信号会降低。所提出的免疫传感器基于层层自组装技术修饰的丝网印刷碳电极,通过广泛的特性研究,成功地通过滴铸 3.0 μL 浓度为 0.1mg mL 的单壁碳纳米管悬浮液,并在 -0.2 V 下沉积 150s 的金纳米粒子来实现传感器的纳米结构化。通过化学修饰将单克隆抗体固定在金纳米粒子表面,最佳浓度为 200μg mL。使用所提出的免疫传感器,在 0.01-10ng mL 的范围内检测到 α- 突触核蛋白,检测限和定量限分别为 4.1 和 12.6pg mL。在胎牛血清中测试浓度的加标回收率为 96.7%、106.2%和 102.9%,同时在一个月内表现出优异的特异性和稳定性。纳米结构免疫传感器为电子转导和生物识别事件提供了良好的界面,实现了 α- 突触核蛋白的快速、敏感和特异性检测,同时基于简单且廉价的技术,需要小体积的样品,这是即时检测应用的关键特征。

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