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金结合肽作为一种选择性层用于 SARS-CoV-2 抗体的电化学检测。

Gold-binding peptide as a selective layer for electrochemical detection of SARS-CoV-2 antibodies.

机构信息

Laboratory of Electrochemical Sensors (LabSensE), Department of Chemistry, Federal University of Paraná (UFPR), CEP 19032, CEP, 81531-980, Curitiba, PR, Brazil; Molecular Biology Laboratory, Graduate Program in Bioprocess Engineering and Biotechnology, Federal University of Paraná (UFPR), CEP, 81531-980, Curitiba, PR, Brazil.

Molecular Biology Laboratory, Graduate Program in Bioprocess Engineering and Biotechnology, Federal University of Paraná (UFPR), CEP, 81531-980, Curitiba, PR, Brazil.

出版信息

Talanta. 2023 May 15;257:124348. doi: 10.1016/j.talanta.2023.124348. Epub 2023 Feb 11.

Abstract

Electrochemical immunosensors are excellent alternatives to prepare portable platforms used for rapid and inexpensive diagnostic of infectious diseases such as the recently emerged COVID-19. Incorporating synthetic peptides as selective recognition layers combined with nanomaterials such as gold nanoparticles (AuNPs) can significantly enhance the analytical performance of immunosensors. In the present study, an electrochemical immunosensor based on solid-binding peptide was built and evaluated towards SARS-CoV-2 Anti-S antibodies detection. The peptide used as recognition site has two important portions: one based on the viral receptor binding domain (RBD), capable of recognizing antibodies of the spike protein (Anti-S), and another suitable for interacting with gold nanoparticles. Gold-binding peptide (Pept/AuNP) dispersion was used directly to modify a screen-printed carbon electrode (SPE). The voltammetric behavior of the [Fe(CN)] probe after every construction and detection step was recorded using cyclic voltammetry by assessing the stability of the Pept/AuNP as a recognition layer onto the electrode surface. Differential pulse voltammetry was used as a detection technique, and a linear working range from 75 ng mL to 15 μg mL was established, with 1.059 μA dec of sensitivity and R = 0.984. The response selectivity against SARS-CoV-2 Anti-S antibodies was investigated in presence of concomitant species. The immunosensor was used to detect SARS-CoV-2 Anti-spike protein (Anti-S) antibodies in human serum samples, successfully differentiating between negative and positive responses of samples at a 95% confidence level. Therefore, the gold-binding peptide is a promising tool to be applied as a selective layer for antibody detection.

摘要

电化学免疫传感器是制备用于快速和廉价诊断传染病(如最近出现的 COVID-19)的便携式平台的绝佳替代品。将合成肽作为选择性识别层与纳米材料(如金纳米粒子(AuNPs))结合使用,可以显著提高免疫传感器的分析性能。在本研究中,构建了一种基于固相结合肽的电化学免疫传感器,并对其进行了评估,用于检测 SARS-CoV-2 抗-S 抗体。用作识别位点的肽具有两个重要部分:一部分基于病毒受体结合域(RBD),能够识别刺突蛋白(Anti-S)的抗体,另一部分适合与金纳米粒子相互作用。金结合肽(Pept/AuNP)分散体直接用于修饰丝网印刷碳电极(SPE)。通过使用循环伏安法评估 Pept/AuNP 作为电极表面上的识别层的稳定性,记录了每一步构建和检测后[Fe(CN)]探针的伏安行为。差分脉冲伏安法被用作检测技术,建立了 75 ng mL 至 15 μg mL 的线性工作范围,灵敏度为 1.059 μA dec,相关系数为 0.984。研究了在共存物质存在下针对 SARS-CoV-2 抗-S 抗体的免疫传感器响应选择性。该免疫传感器用于检测人血清样本中的 SARS-CoV-2 抗刺突蛋白(Anti-S)抗体,在 95%置信水平下成功区分了阴性和阳性样本的反应。因此,金结合肽是一种有前途的工具,可作为用于抗体检测的选择性层。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1751/9918321/0513da7f0be0/ga1_lrg.jpg

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