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基于合成的 rGO/MoS/AgNPs 纳米复合材料的无标记电化学免疫传感测定胶质纤维酸性蛋白(GFAP)。在人脑脊液中的测定应用。

Label-free electrochemical immunosensing of glial fibrillary acidic protein (GFAP) at synthesized rGO/MoS/AgNPs nanocomposite. Application to the determination in human cerebrospinal fluid.

机构信息

Department of Analytical Chemistry, Faculty of Chemistry, University Complutense of Madrid, 28040, Madrid, Spain.

Department of Analytical Chemistry, Faculty of Chemistry, University Complutense of Madrid, 28040, Madrid, Spain.

出版信息

Talanta. 2024 Apr 1;270:125597. doi: 10.1016/j.talanta.2023.125597. Epub 2023 Dec 23.

Abstract

An electrochemical bioplatform involving screen-printed carbon electrodes modified with rGO/MoS/AgNPs nanocomposites, the covalent immobilization of the specific capture antibody, and label-free detection has been developed for the determination of Glial Fibrillary Acidic Protein (GFAP). The resulting immunosensor profits the benefits of the rGO high conductivity, the pseudo-peroxidase activity of MoS and the electrocatalytic effect provided by AgNPs for improving the reduction current responses of hydrogen peroxide at the electrode surface. GFAP is a biomarker of central nervous system injuries has been proposed for the detection and monitoring of neurological diseases as epilepsy, encephalitis, or multiple sclerosis. For the first time, amperometric detection of the immunosensing event was performed by measuring the electrocatalytic response of hydrogen peroxide reduction at the modified electrode. Several techniques including scanning (SEM) and transmission (TEM) electron microscopies were used for the characterization of the synthesized composite whilst electrochemical impedance spectroscopy (EIS) using the redox probe Fe(CN) was employed to evaluate the success of the steps implied in the fabrication of the immunosensor. After optimization of the involved experimental variables, a linear calibration plot for GFAP was constructed over the 0.6-100 ng mL range, and a detection limit of 0.16 ng mL was achieved. The developed immunosensor was successfully applied to the determination of GFAP in human cerebrospinal fluid (CSF) of patients diagnosed with encephalitis.

摘要

一种电化学生物平台,涉及用 rGO/MoS/AgNPs 纳米复合材料修饰的丝网印刷碳电极,通过共价固定特异性捕获抗体和无标记检测,用于测定神经胶质纤维酸性蛋白 (GFAP)。所得到的免疫传感器得益于 rGO 的高导电性、MoS 的类过氧化物酶活性以及 AgNPs 提供的电催化效应,从而改善了电极表面过氧化氢的还原电流响应。GFAP 是中枢神经系统损伤的生物标志物,已被提议用于检测和监测癫痫、脑炎或多发性硬化症等神经疾病。首次通过测量修饰电极上过氧化氢还原的电催化响应来进行免疫传感事件的电流检测。使用扫描 (SEM) 和透射 (TEM) 电子显微镜等几种技术对合成的复合材料进行了表征,而电化学阻抗谱 (EIS) 使用氧化还原探针 Fe(CN) 来评估免疫传感器制造过程中所涉及的步骤的成功。在优化了所涉及的实验变量后,在 0.6-100ngmL 的范围内构建了用于 GFAP 的线性校准图,并且实现了 0.16ngmL 的检测限。该开发的免疫传感器成功地应用于测定脑炎患者的人脑脊液 (CSF) 中的 GFAP。

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