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

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.

Keywords

• Adenosine deaminase acting on RNA1 (ADAR1)
• Adenosine-to-inosine (A-to-I) RNA editing
• Aptamer
• DEPSOR
• Differential pulse voltammetry (DPV)
• Cancer prognosis