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Ligand-Based Stability Changes in Duplex DNA Measured with a Microscale Electrochemical Platform

1
Department of Chemistry and Biochemistry, University of Maryland, College Park, MD 20742, USA
2
Department of Chemistry, Institute for Drug Discovery, Purdue University, West Lafayette, IN 47907, USA
3
Biomolecular Measurement Division, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA
*
Authors to whom correspondence should be addressed.
Now at Spectrix Analytical Services, LLC, North Haven, CT 06473, USA.
Biosensors 2019, 9(2), 54; https://doi.org/10.3390/bios9020054
Received: 12 March 2019 / Revised: 3 April 2019 / Accepted: 9 April 2019 / Published: 12 April 2019
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Abstract

Development of technologies for rapid screening of DNA secondary structure thermal stability and the effects on stability for binding of small molecule drugs is important to the drug discovery process. In this report, we describe the capabilities of an electrochemical, microdevice-based approach for determining the melting temperatures (Tm) of electrode-bound duplex DNA structures. We also highlight new features of the technology that are compatible with array development and adaptation for high-throughput screening. As a foundational study to exhibit device performance and capabilities, melting-curve analyses were performed on 12-mer DNA duplexes in the presence/absence of two binding ligands: diminazene aceturate (DMZ) and proflavine. By measuring electrochemical current as a function of temperature, our measurement platform has the ability to determine the effect of binding ligands on Tm values with high signal-to-noise ratios and good reproducibility. We also demonstrate that heating our three-electrode cell with either an embedded microheater or a thermoelectric module produces similar results. The ΔTm values we report show the stabilizing ability of DMZ and proflavine when bound to duplex DNA structures. These initial proof-of-concept studies highlight the operating characteristics of the microdevice platform and the potential for future application toward other immobilized samples. View Full-Text
Keywords: DNA; electrochemical sensor; ligand-based stabilization; melting profiles; microfabrication; microheater; rapid temperature control; square wave voltammetry DNA; electrochemical sensor; ligand-based stabilization; melting profiles; microfabrication; microheater; rapid temperature control; square wave voltammetry
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Robinson, S.M.; Shen, Z.; Askim, J.R.; Montgomery, C.B.; Sintim, H.O.; Semancik, S. Ligand-Based Stability Changes in Duplex DNA Measured with a Microscale Electrochemical Platform. Biosensors 2019, 9, 54.

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