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实时液相生物分子传感和识别的新方法。该系统依赖于微流体传感平台中一系列生物纳米孔中离子传输的随机测量，实现了紧凑、低成本和自动化的单分子质量测量。微流控系统将单个离子通道传感元件的多路复用阵列嵌入在一个一次性热塑性微流控芯片内的离散双层脂质膜中，多层通道能够将分析物动态输送到传感部位。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标筛选的应用，允许大幅提高分析通量，而无需由训练有素的人员手动操作。更重要的是，该系统将利用我们团队最近展示的结果，在分子量的基础上对单个分子进行鉴定和定量。测量将实时进行，在水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的灌注到多路传感位点。由此产生的平台将是一种独特的使能技术，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。

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.