EWOD-MALDI-MS for High-Throughput Proteomics

蛋白质组学研究需要能够收集、储存、分类和分析大量蛋白质的方法和仪器。 大量的信息。为了加快蛋白质组学研究的进展，我们建议开发 基于电介质上电润湿（EWOD）的微流体，这将使样品处理步骤能够 与用于基质辅助激光解吸/电离质谱的高通量阵列的创建相结合 质谱法（MALDI-MS）。在EWOD中，电场用于操纵液体表面张力。通过 通过在电介质层下制造的电极上施加一系列电势， 从水库中产生，移动到特定的位置，加入，混合和分裂。基于EWOD的微流控 装置是可重新配置的，并且可以处理水中或水中的中性和带电分析物和颗粒， 有机溶剂本研究计画包含五个目标：1）EWOD装置的设计与制作; 2） EWOD作为MALDI-MS的样品制备工具的表征; 3）基于EWOD的样品纯化 4）EWOD-MALDI-MS与微型HPLC和CD分离的集成，使用新的 方法，“代客泊车MALDI”，和5）EWOD-MALDI-MS的两个应用：（a）疾病检测 唾液中的标志物，和（B）产生用于功能蛋白质组学的蛋白质阵列。因为它有潜力 为了将许多样品处理步骤与MALDI-MS的高通量阵列的创建相结合， 所提出的方法将代表现有蛋白质组学技术的重大进步。更广泛地说， 我们开发的基于EWOD的设备和样品处理协议将适用于广泛的 生物分析化学和化学生物学中的基于阵列的方法。

Proteomics research requires methods and instruments that can collect, store, catalogue and analyze vast amounts of information. To accelerate progress in proteomics research, we propose to develop electrowetting-on-dielectric (EWOD)-based microfluidics that will enable sample processing steps to be integrated with creation of high-throughput arrays for matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS). In EWOD, electric fields are used to manipulate liquid surface tension. By applying a sequence of potentials across electrodes fabricated under a dielectric layer, droplets can be created from a reservoir, moved to specific locations, joined, mixed and divided. EWOD-based microfluidic devices are reconfigurable, and can handle neutral and charged analytes and particulates, in water or organic solvents. The research plan addresses five objectives: 1) EWOD device design and fabrication; 2) chracterization of EWOD as a sample preparation tool for MALDI-MS; 3) EWOD-based sample purification and processing; 4) integration of EWOD-MALDI-MS with micro-HPLC and CD separations, using a new method, "valet parking MALDI," and 5) two applications of EWOD-MALDI-MS: (a) detection of disease markers in saliva, and (b) generation of protein arrays for functional proteomics. Because it has the protential to integrate many sample processing steps with creation of high-throughput arrays for MALDI-MS, the proposed method will represent a major advance over existing technologies for proteomics. More broadly, the EWOD-based devices and sample processing protocols we develop will be applicable in a wide range of array-based methods in bioanalytical chemistry anad chemical biology.