LC-MICRODISPENSER FOR HIGH RESOLUTION MALDI ANALYSIS

我们将开发一种基于喷墨的微点胶系统，具有集成的液相色谱功能。该系统将用于制备用于蛋白质组学分析的基质辅助激光解吸/电离（MALDI）质谱靶。MicroFab的高速“打印机”将把消化后的蛋白质样品上的多个斑点沉积到金属板上。阵列中的每个点本质上都是肽图谱的片段收集容器。金属板将直接放入MALDI质谱仪中进行分析。由于组分直接放到MALDI网格上，分析与液相色谱-质谱（LC/MS）一样灵敏，而不会牺牲MALDI的高通量特性。在第一阶段，我们将展示原理证明，使用MicroFab点胶装置和集成的反相树脂微柱来分配肽微点，选择一种形成良好晶体的质谱基质，并确定系统的灵敏度。在第二阶段，我们将建立一个原型打印机，能够在MALDI目标上生产肽片段阵列，用于蛋白质组学应用。阵列生产微分配器将用于并行处理，产生更高的吞吐量。MALDI分析具有固有的高通量，提高灵敏度将大大增加蛋白质组学研究的成果。建议的商业应用：基于蛋白质组学的药物发现、疾病诊断和疾病监测研究需要大规模并行处理。为了继续进行这些研究，需要一种高通量且非常敏感的仪器。我们的系统将成为该领域的先驱，并能够占据相当大的市场份额。

We will develop an ink-jet based micro-dispensing system with integrated liquid chromatography capability. This system will be used to prepare matrix- assisted laser desorption/ionization (MALDI) mass spectrometry targets for use in proteomic analysis. MicroFab's high-speed "printer" will deposit multiple spots from a digested protein sample onto a metal plate. Each spot in the array will essentially be the fraction collection vessel for peptide mapping. The metal plate will be put directly into the MALDI mass spectrometer for analysis. Since the fractions are put directly onto the MALDI grid, analysis is as sensitive as liquid chromatography-mass spectrometry (LC/MS) without sacrificing the high- throughput characteristic of MALDI. In Phase I we will demonstrate the proof of principle by dispensing peptide microspots with a MicroFab dispensing device with integrated reversed-phase resin microcolumn, selecting a MS matrix for good crystal formation, and determining the sensitivity of the system. In Phase II we will build a prototype printer capable of producing peptide fraction arrays on MALDI targets for proteomic applications. An array-producing microdispenser will be utilized for parallel processing, yielding even higher throughput. MALDI analysis is inherently high- throughput and increasing the sensitivity will drastically increase the output ofproteomic research efforts. PROPOSED COMMERCIAL APPLICATIONS: Proteomic-based research for drug discovery, diagnosing disease and monitoring disease requires massively parallel processing. A high-throughput instrument which is also very sensitive is required for these studies to continue. Our system would be a pioneer in the field and able to capture a significant share of the market.