EWOD-MALDI-MS for High-Throughput Proteomics

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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.