Multichannel Surface Plasmon Resonance for Quantitative Biomolecular Kinetics

该奖项支持开发一种新的仪器，通过使用基于全光学表面等离子体共振(SPR)的方法来测量生物分子结合的实时定量动力学。现有的基于SPR的仪器通常用于研究聚合物与其他分子的相互作用，包括小配体和蛋白质。该仪器将采用多通道生物传感器阵列格式，并使用多波长激励和角度扫描能力来优化灵敏度和时间响应。与现有的SPR仪器相比，PI实验室以前开发的新的SPR方法有望提供更高的灵敏度和对未标记生物分子的覆盖范围。将开发的用于数据分析的软件将包括使用针对多个测量的自洽全局分析方法对缔合和解离动力学的详细建模，以及PI的实验室还开发了新的动力学模型。将开发的仪器提供对16个不同通道(4个独立流动单元)的结合动力学的同时监测，时间响应预计在1毫秒范围内。合理选择激发和检测频率将充分利用正在研究的特定配体的光学特性；这有望提供比目前基于SPR的生物传感器高1到2个数量级的额外灵敏度

This award supports the development of a new instrument for measurement of real-time quantitative kinetics of biomolecular binding through use of all-optical surface plasmon resonance (SPR)-based methods. Existing SPR-based instrumentation is commonly used to study interactions of polymers and other molecules, including small ligands and proteins. The proposed instrument will employ a multi-channel biosensor array format, and use multi-wavelength excitation and angular scanning capabilities to optimize sensitivity and time response. Novel SPR methods, developed previously in the PI's laboratory, are expected to provide far greater sensitivity and improved coverage for unlabeled biomolecules in comparison to existing SPR instruments. Software to be developed for data analysis will incorporate detailed modeling of association and dissociation kinetics using self-consistent global analysis methods for multiple measurements and new kinetic models also developed in the PI's laboratory.The instrument to be developed provides simultaneous monitoring of binding kinetics for 16 different channels (4 independent flow cells) with a time response expected to be in the range of 1 millisecond. Judicious choice of excitation and detection frequencies will take full advantage of the optical characteristics of the particular ligands under study; this is expected to provide an additional 1 to 2 orders of magnitude higher sensitivity than current SPR-based biosensors