Department of Chemistry and Biochemistry, University of Maryland, College Park, MD 20742, USA.
Department of Chemistry, Institute for Drug Discovery, Purdue University, West Lafayette, IN 47907, USA.
Biosensors (Basel). 2019 Apr 12;9(2):54. doi: 10.3390/bios9020054.
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 () 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 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 Δ 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.
快速筛选 DNA 二级结构热稳定性的技术的发展及其对小分子药物结合稳定性的影响对药物发现过程至关重要。在本报告中，我们描述了一种基于电化学的微器件方法在确定电极结合的双链 DNA 结构的熔点（）方面的能力。我们还突出了该技术的新特点，这些特点与阵列开发和高通量筛选的适应性兼容。作为展示设备性能和能力的基础研究，在存在/不存在两种结合配体的情况下对 12 聚体 DNA 双链进行了熔融曲线分析：二甲苯胺脒（DMZ）和吖啶黄。通过测量电化学电流随温度的变化，我们的测量平台具有以高信噪比和良好重现性确定结合配体对 值影响的能力。我们还证明，使用嵌入式微加热器或热电模块加热我们的三电极池会产生类似的结果。我们报告的 Δ 值表明 DMZ 和吖啶黄与双链 DNA 结构结合时的稳定能力。这些初步的概念验证研究突出了微器件平台的操作特性及其在未来对其他固定化样品的应用潜力。
