Exonuclease I-assisted General Strategy to Convert Aptamer-based Electrochemical Biosensors from "Signal-off" to "Signal-On".

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2020
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Abstract
In terms of how the signal varies in response to increased concentration of an analyte, sensors can be classified as either "signal-on" or "signal-off" format. While both types hold potentials to be sensitive, selective, and reusable, in many situations "signal-on" sensors are preferred as of their low background signal and better selectivity. In this study, with the detection of lysozyme using its DNA aptamer as a trail system, for the first time we demonstrated that such an aptamer-based electrochemical biosensor can be converted from intrinsically "signal-off" to "signal-on" with the aid of a DNA exonuclease. The fact that the step-wise cleavage of anti-lysozyme aptamer catalyzed by Exonuclease I (Exo I) is entirely inhibited upon binding lysozyme, leads to the selective removal of unbound DNA probes (thiolate anti-lysozyme DNA aptamer strands immobilized on gold electrode) upon the introduction of Exo I to the sensor. With the aid of electrostatically bound redox cations ([Ru(NH3)6]3+), we were able to quantitate the number of aptamer strands that are bound with lysozymes via conventional cyclic voltammetry (CV) measurements. We demonstrated that Exo I-assisted signal-on conversion protocol not only improves the sensing performance (10 times better limit of detection), but also promises a versatile strategy for DNA-based biosensor design, i.e., it can be readily adapted to other aptamer-protein binding systems (thrombin, as another example).
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gao2020exonucleaseanalytical Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Gao, Xiaoyi;Qi, Lin;Liu, Kun;Meng, Chenchen;Li, Yunchao;Yu, Hua-Zhong;
Journal Analytical chemistry
Year 2020
DOI
10.1021/acs.analchem.0c00005
URL
Keywords
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