Ligand-Based Stability Changes in Duplex DNA Measured with a Microscale Electrochemical Platform
AbstractDevelopment 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
- Supplementary File 1:
PDF-Document (PDF, 280 KB)
Share & Cite This Article
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.
Robinson SM, Shen Z, Askim JR, Montgomery CB, Sintim HO, Semancik S. Ligand-Based Stability Changes in Duplex DNA Measured with a Microscale Electrochemical Platform. Biosensors. 2019; 9(2):54.Chicago/Turabian Style
Robinson, Sarah M.; Shen, Zuliang; Askim, Jon R.; Montgomery, Christopher B.; Sintim, Herman O.; Semancik, Steve. 2019. "Ligand-Based Stability Changes in Duplex DNA Measured with a Microscale Electrochemical Platform." Biosensors 9, no. 2: 54.
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.