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了解生物化学过程的基本任务 生物体，发现生物标记物和开发药物。当今主流的商业检测技术 是基于对质量变化的测量，其难以检测到小分子(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，蛋白质磷酸化 和其他翻译后修改)。然而，这些分子和反应对生物学来说是至关重要的 功能，以及疾病的发生和发展。小分子也占FDA批准的90%以上 毒品。 为了解决这一未得到满足的需求，本项目旨在开发一种纳米振荡器阵列(NOA)检测技术。 每个纳米振荡器由一个绑在表面等离子体共振(SPR)传感器表面的纳米颗粒组成 有一个灵活的分子连接物。纳米颗粒被目标蛋白预先功能化。在应用 在垂直于传感器表面的交变电场下，纳米颗粒被强迫振荡，并且振荡 幅度是用亚纳米分辨率的SPR成像方法测量的。此分辨率对应于 电子电荷的一小部分，使NoA对分子与蛋白质上的结合特别敏感 纳米颗粒，或通过改变净电荷或电荷对蛋白质进行翻译后修饰 蛋白质的分布。生物传感仪器公司、亚利桑那州立大学、 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，团队将共同为NoA的商业化做好准备，i)扩展 NOA作为一种新的商业化小分子定量技术的初步和可行性研究 结合和生化反应，二)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、工作流程和 特定于应用程序的工具，以及iii)执行验证测试并展示启用NoA的案例研究 应用程序(即杀手级应用程序)。该项目的成功将导致一种新技术来解决 对量化小分子结合动力学和生化反应动力学的需求尚未得到满足。

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.