A novel aptamer-antibody sandwich electrochemical sensor for detecting ADAR1 in complex biological samples

IF 10.61 Q3 Biochemistry, Genetics and Molecular Biology

Biosensors and Bioelectronics: X Pub Date : 2024-05-16 DOI:10.1016/j.biosx.2024.100491

Madhu Biyani , Kirti Sharma , Maeda Shoei , Hinako Akashi , Masataka Nakano , Miki Nakajima , Manish Biyani

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引用次数: 0

Abstract

Human adenosine deaminase acting on RNA1 (ADAR1) is an adenosine-to-inosine (A-to-I) RNA-editing enzyme involved in various types of cancer progression. ADAR1 has emerged as a novel prognostic biomarker for cancer. This study describes the application of a newly identified 70-nt DNA aptamer (Apt38483) against ADAR1 to develop a portable and simple electrochemical biosensor platform for the rapid and sensitive detection of ADAR1 in cell lysates. We selected an ADAR1-specific DNA aptamer from a randomized 70-nt single-stranded DNA library using a competitive in vitro selection method. ADAR1 in the cell lysate was sandwiched onto a bare carbon working electrode of an electro-chemically printed chip between the ADAR1 antibody and gold nanoparticles (40 nm) conjugated with Apt38483, followed by electrochemical analysis using differential pulse voltammetry (DPV) for sensor demonstration. A highly sensitive change in current was observed for as little as 0.53 nM ADAR1 in human embryonic kidney cell lysate. Thus, the merging of a novel DNA aptamer probe for ADAR1 with an electrochemical transduction method enabled the development of a simple, low-cost, and rapid method for the direct measurement of ADAR1 in cell lysates and indicated great potential for the development of an ADAR1 analysis platform, which would be useful in cancer prognosis.

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用于检测复杂生物样品中 ADAR1 的新型适配体-抗体夹心电化学传感器

作用于 RNA1 的人类腺苷脱氨酶（ADAR1）是一种腺苷转化为肌苷（A-to-I）的 RNA 编辑酶，与各种癌症进展有关。ADAR1 已成为一种新型的癌症预后生物标志物。本研究介绍了应用新发现的针对 ADAR1 的 70-nt DNA 类似物（Apt38483）开发便携式简易电化学生物传感器平台，用于快速灵敏地检测细胞裂解物中的 ADAR1。我们采用竞争性体外选择方法，从随机 70-nt 单链 DNA 文库中筛选出了 ADAR1 特异性 DNA 类似物。将细胞裂解液中的 ADAR1 夹在电化学印刷芯片的裸炭工作电极上，在 ADAR1 抗体和与 Apt38483 共轭的金纳米粒子（40 nm）之间，然后使用差分脉冲伏安法（DPV）进行电化学分析，以演示传感器。在人类胚胎肾细胞裂解物中，只要 0.53 nM ADAR1 就能观察到高灵敏度的电流变化。因此，将 ADAR1 的新型 DNA 类似物探针与电化学转导方法相结合，开发出了一种简单、低成本、快速的直接测量细胞裂解物中 ADAR1 的方法，这表明 ADAR1 分析平台的开发具有巨大潜力，将有助于癌症预后。

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来源期刊

Biosensors and Bioelectronics: X Biochemistry, Genetics and Molecular Biology-Biophysics

CiteScore

4.60

自引率

0.00%

发文量

166

审稿时长

54 days

期刊介绍： Biosensors and Bioelectronics: X, an open-access companion journal of Biosensors and Bioelectronics, boasts a 2020 Impact Factor of 10.61 (Journal Citation Reports, Clarivate Analytics 2021). Offering authors the opportunity to share their innovative work freely and globally, Biosensors and Bioelectronics: X aims to be a timely and permanent source of information. The journal publishes original research papers, review articles, communications, editorial highlights, perspectives, opinions, and commentaries at the intersection of technological advancements and high-impact applications. Manuscripts submitted to Biosensors and Bioelectronics: X are assessed based on originality and innovation in technology development or applications, aligning with the journal's goal to cater to a broad audience interested in this dynamic field.