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能力。该系统将用于制备基质辅助激光解吸/电离(MALDI)质谱靶，用于蛋白质组分析。MicroFab的高速“打印机”可以将消化的蛋白质样本中的多个斑点沉积到金属板上。阵列中的每个点基本上都是用于肽图谱的组分收集容器。金属板将被直接放入MALDI质谱仪进行分析。由于馏分被直接放入MALDI网格，因此分析与液-质联用(LC/MS)一样灵敏，而不会牺牲MALDI的高通量特性。在第一阶段，我们将通过使用集成了反相树脂微柱的MicroFab分配设备来分配多肽微点，选择形成良好晶体的MS基质，并确定系统的灵敏度，从而证明原理的正确性。在第二阶段，我们将建造一台原型打印机，能够在MALDI靶标上产生用于蛋白质组学应用的多肽部分阵列。阵列生产微分配器将用于并行处理，从而产生更高的吞吐量。MALDI分析本质上是高通量的，提高灵敏度将极大地增加蛋白质组研究的产出。拟议的商业应用：基于蛋白质组的药物发现、疾病诊断和疾病监测研究需要大规模的并行处理。要继续进行这些研究，需要一台同样非常灵敏的高通量仪器。我们的系统将是该领域的先驱，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.