Ligand-based stability changes in duplex DNA measured with a microscale electrochemical platform

Published

April 12, 2019

Author(s)

Sarah M. Robinson, Zuliang Shen, Jon R. Askim, Christopher B. Montgomery, Herman O. Sintim, Stephen Semancik

Abstract

Development of technologies for rapidly screening the thermal stability of DNA secondary structures 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 analysis was performed on 12-mer DNA duplexes in the presence/absence of two binding ligands: diminazene aceturate (DMZ) and proflavine. Results show that our measurement platform has the ability to measure, from current vs. temperature traces, the effect of binding ligands on Tm values with high signal-to-noise ratios and good reproducibility. We also demonstrate that our 3-electrode cell can be heated by either an embedded microheater or with a thermoelectric module, producing similar results. The ΔTm values we report show the stabilizing ability of DMZ and proflavine when bound to DNA duplex structures. These initial proof-of-concept studies highlight the operating characteristics of the microdevice platform and the potential future application toward other immobilized samples.

Citation

Biosensors

Volume

9

Issue

2

Pub Type

Journals

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DOI Link

Keywords

DNA, microheater, microfabrication, melting profiles, rapid temperature control, ligand-based stabilization, Square Wave Voltammetry

Citation

Robinson, S. , Shen, Z. , Askim, J. , Montgomery, C. , Sintim, H. and Semancik, S. (2019), Ligand-based stability changes in duplex DNA measured with a microscale electrochemical platform, Biosensors, [online], https://doi.org/10.3390/bios9020054 (Accessed November 23, 2024)