Single Molecule Mass Spectrometry in a Microfluidic Nanopore Chip

描述（由申请人提供）：将研究一种在单分子水平上进行实时液相生物分子传感和鉴别的新方法。该系统依赖于跨微流体传感平台中的生物纳米孔阵列的离子传输的随机测量，从而能够以紧凑、低成本和自动化的形式进行单分子质量测量。微流体系统将联合收割机组合嵌入一次性热塑性微流体芯片内的离散双层脂质膜中的单个离子通道感测元件的多路复用阵列，其中多层通道能够将分析物动态递送到感测位点。在其最简单的形式中，微流控芯片将提供传统电生理仪器的替代方案，用于从基本离子通道研究到药物靶点筛选的应用，从而允许显著更高的分析通量，而无需训练有素的人员进行手动操作。更重要的是，该系统将利用我们团队最近展示的结果，根据分子量对单个分子进行识别和定量。测量将实时发生，在水性环境中提供时间分辨的原位分析，而不需要气相电离，直接控制分析物和其他试剂到多路复用传感位点的灌注。由此产生的平台将是一个独特的使能技术，用于广泛的生物分子分析，并将被证明用于复杂样品中肽的鉴定和定量。) 公共卫生相关性：生物传感器平台能够在单个分子水平上基于分子质量来区分分子，这为推进我们对生物过程的基本理解提供了重要的希望。该项目致力于开发一种独特的微流体支持平台，该平台将允许对复杂样品中的单个生物分子进行基于质量的识别，为了解疾病状态和进展的分子网络提供新的窗口。

DESCRIPTION (provided by applicant): A new approach to real-time liquid-phase biomolecular sensing and discrimination at the single molecule level will be investigated. The system relies on stochastic measurements of ion transport across an array of biological nanopores in a microfluidic sensing platform, enabling single molecule mass measurements in a compact, low cost, and automated format. The microfluidic system will combine multiplexed arrays of individual ion channel sensing elements embedded in discrete bilayer lipid membranes within a disposable thermoplastic microfluidic chip, with multilayer channels enabling the dynamic delivery of analytes to the sensing sites. In its simplest form, the microfluidic chip will provide an alternative to traditional electrophysiological instruments for applications ranging from fundamental ion channel studies to drug target screening, allowing substantially higher analytical throughput without the need for manual operation by highly trained personnel. More significantly, the system will leverage recent results demonstrated by our team towards the identification and quantification of individual molecules on the basis of their molecular weight. Measurements will occur in real-time, providing time-resolved in-situ analysis within an aqueous environment without the need for gas-phase ionization, with direct control over the perfusion of analytes and other reagents to the multiplexed sensing sites. The resulting platform will be a unique enabling technology for a broad range of biomolecular analyses, and will be demonstrated for the identification and quantification of peptides within complex samples. ) PUBLIC HEALTH RELEVANCE: Biosensor platforms capable of discriminating molecules on the basis of their molecule masses at the level of individual molecules offer significant promise towards advancing our fundamental understanding of biological processes. This project addresses the development of a unique microfluidic-enabled platform that will allow the mass-based identification of individual biomolecules within complex samples, providing a new window into the molecular networks that underlie disease state and progression.