Nano-Oscillator Arrays for Sensitive Plasmonic Detection of Molecular Interactions and Reactions

摘要 检测分子间的相互作用和反应是理解生命中生物化学过程的基本任务 发现生物标志物和开发药物。当今主流的商业检测技术 是基于测量质量变化，其难以检测小分子（例如，代谢物，激素， 和神经递质），以及涉及小质量变化的生化反应（例如，蛋白磷酸化 和其它翻译后修饰）。然而，这些分子和反应对生物学至关重要。 功能以及疾病的发生和进展。小分子也占FDA批准的药物的90%以上。 毒品 为了解决这一未满足的需求，该项目旨在开发纳米振荡器阵列（NOA）检测技术。 每个纳米振荡器由一个纳米粒子拴在表面等离子体共振（SPR）传感器表面 带有柔性分子连接体。纳米颗粒用靶蛋白预官能化。当应用 在垂直于传感器表面的交变电场中，纳米颗粒被迫振荡，并且振荡 振幅用亚纳米分辨率的SPR成像方法测量。该决议对应于一个 电子电荷的分数，使得NOA对分子与蛋白质的结合特别敏感， 纳米颗粒，或通过改变净电荷或电荷的蛋白质的翻译后修饰 蛋白质的分布。Biosensing Instrument，Inc.亚利桑那州立大学， 和制药公司已经产生了大量的初步数据，证明了强大的 NOA的潜力 在这个快速通道项目中，该团队将共同努力，通过I）扩展 NOA作为一种新的商业化小分子定量技术的初步可行性研究 结合和生物化学反应，II）开发商业原型NOA系统，包括光学 仪器，信号处理算法，NOA传感器生产方法，工作流程，以及 应用程序特定的工具，和III）进行验证测试，并显示NOA启用的案例研究 杀手级应用（Killer Application）。该项目的成功将导致一种新的技术来解决 对定量小分子结合动力学和生化反应动力学的未满足的需求。

ABSTRACT Detecting molecular interactions and reactions are basic tasks to understand biochemical processes in living organisms, discover biomarkers and develop drugs. Today's mainstream commercial detection technologies are based on measuring mass changes, which struggle to detect small molecules (e.g., metabolites, hormones, and neurotransmitters), and biochemical reactions involving small mass changes (e.g., protein phosphorylation and other post-translational modifications). However, these molecules and reactions are critical to biological functions, and disease initiation and progression. Small molecules also count for over 90% of FDA approved drugs. To address this unmet need, this project aims to develop a nano-oscillator array (NOA) detection technology. Each nano-oscillator consists of a nanoparticle tethered to a surface plasmon resonance (SPR) sensor surface with a flexible molecular linker. The nanoparticle is pre-functionalized with target proteins. When applying an alternating electric field normal to the sensor surface, the nanoparticle is forced to oscillate, and the oscillation amplitude is measured with a SPR imaging method with sub-nm resolution. This resolution corresponds to a fraction of electron charge, making NOA particularly sensitive to the binding of molecules to the proteins on the nanoparticle, or post-translational modification of the proteins via a change in the net charge or charge distribution of the proteins. Collaborative efforts among Biosensing Instrument, Inc., Arizona State University, and pharmaceutical companies have resulted in substantial preliminary data that demonstrates the powerful potential of NOA. In this fast track project, the team will work together to prepare NOA for commercialization by I) expanding on preliminary and feasibility studies of NOA as a new commercial technology for quantifying small molecule binding and biochemical reactions, II) developing a commercial prototype NOA system, including optical instrumentation, signal processing algorithms, NOA sensor production methods, workflow processes, and application specific tools, and III) carrying out both validation tests and show case studies on NOA enabled applications (i.e. killer applications). The success of this project will lead to a new technology to address the unmet need for quantifying small molecule binding kinetics and biochemical reactions kinetics.