Paul Ananya, Musetti Caterina, Nanjunda Rupesh, Wilson W David
Department of Chemistry, Center for Diagnostics and Therapeutics, Georgia State University, Atlanta, GA, USA.
Department of Screening, Profiling and Mechanistic Biology, Platform Technology and Science, Glaxo Smith Kline, Collegeville, PA, USA.
Methods Mol Biol. 2019;2035:63-85. doi: 10.1007/978-1-4939-9666-7_4.
Biosensor-surface plasmon resonance (SPR) technology is now well established as a quantitative approach for the study of nucleic acid interactions in real time, without the need for labeling any components of the interaction. The method provides real-time equilibrium and kinetic characterization for quadruplex DNA interactions and requires small amounts of materials and no external probe. A detailed protocol for quadruplex-DNA interaction analyses with a variety of binding molecules using biosensor-SPR methods is presented. Explanations of the SPR method with basic fundamentals for use and analysis of results are described with recommendations on the preparation of the SPR instrument, sensor chips, and samples. Details of experimental design, quantitative and qualitative data analyses, and presentation are described. Some specific examples of small molecule-DNA quadruplex interactions are presented with results evaluated by both kinetic and steady-state SPR methods.
生物传感器表面等离子体共振（SPR）技术现已成为一种成熟的定量方法，用于实时研究核酸相互作用，无需对相互作用的任何组分进行标记。该方法为四链体DNA相互作用提供实时平衡和动力学表征，所需材料量少且无需外部探针。本文介绍了使用生物传感器SPR方法对四链体DNA与多种结合分子相互作用进行分析的详细方案。文中描述了SPR方法的基本原理、使用方法和结果分析，并给出了SPR仪器、传感器芯片和样品制备的建议。还介绍了实验设计、定量和定性数据分析以及结果呈现的细节。文中给出了小分子与DNA四链体相互作用的一些具体例子，并通过动力学和稳态SPR方法对结果进行了评估。
