Competitive electrochemical immunosensors by immobilization of hexahistidine-rich recombinant proteins on the signal labels

IF 4.5 3区化学 Q1 Chemical Engineering

Journal of Electroanalytical Chemistry Pub Date : 2023-09-01 DOI:10.1016/j.jelechem.2023.117662

Yong Chang , Meiling Liu , Tong Wu , Ruting Lin , Lin Liu , Qijun Song

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引用次数: 1

Abstract

Most of proteins in the commercial ELISA kits are recombinant with hexahistidine (His₆) tags. Based on this fact, we proposed a simple procedure for the preparation of signal labels and the design of competitive immunosensors. The signal labels were fabricated through the high-affinity interaction between the His₆ tails on the surface of recombinant proteins and the coordinatively unsaturated copper ions on the surface of pristine Cu-based metal organic framework (Cu-MOF). The recombinant protein-modified MOF could be captured by the sensor electrode, thus producing a strong differential pulse voltammetry signal through the electrochemical reduction of Cu²⁺ ions in MOF. However, the target protein in sample solution could compete with the His₆-tagged protein on the surface of Cu-MOF to bind the antibody attached on the electrode, thus leading to the decrease in the electrochemical signal. The peak current showed an inversely relationship with the concentration of target protein. The competitive immunosensor exhibited a linear range of 1 pg/mL to 1 ng/mL with SARS-CoV-2 nucleocapsid protein (N-protein) as the target analyte. We believe that the method can be used to detect other proteins by utilizing the characteristic of recombinant proteins in commercial kits.

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通过将富含六组氨酸的重组蛋白固定在信号标签上的竞争性电化学免疫传感器

商用ELISA试剂盒中的大多数蛋白质都带有六组氨酸(His6)标签。基于这一事实，我们提出了一种简单的制备信号标签和竞争性免疫传感器的方法。通过重组蛋白表面的His6尾部与原始cu基金属有机骨架(Cu-MOF)表面的配位不饱和铜离子之间的高亲和相互作用制备信号标签。重组蛋白修饰的MOF可以被传感器电极捕获，从而通过电化学还原MOF中的Cu2+离子产生强烈的差分脉冲伏安信号。然而，样品溶液中的靶蛋白会与Cu-MOF表面的his6标记蛋白竞争，与附着在电极上的抗体结合，从而导致电化学信号减弱。峰值电流与靶蛋白浓度呈反比关系。竞争性免疫传感器以SARS-CoV-2核衣壳蛋白(n蛋白)为目标分析物，线性范围为1 pg/mL ~ 1 ng/mL。我们相信该方法可以利用商业试剂盒中重组蛋白的特性来检测其他蛋白质。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Electroanalytical Chemistry Chemical Engineering-General Chemical Engineering

CiteScore

7.50

自引率

6.70%

发文量

912

审稿时长

>12 weeks

期刊介绍： The Journal of Electroanalytical Chemistry is the foremost international journal devoted to the interdisciplinary subject of electrochemistry in all its aspects, theoretical as well as applied. Electrochemistry is a wide ranging area that is in a state of continuous evolution. Rather than compiling a long list of topics covered by the Journal, the editors would like to draw particular attention to the key issues of novelty, topicality and quality. Papers should present new and interesting electrochemical science in a way that is accessible to the reader. The presentation and discussion should be at a level that is consistent with the international status of the Journal. Reports describing the application of well-established techniques to problems that are essentially technical will not be accepted. Similarly, papers that report observations but fail to provide adequate interpretation will be rejected by the Editors. Papers dealing with technical electrochemistry should be submitted to other specialist journals unless the authors can show that their work provides substantially new insights into electrochemical processes.