Microfluidic Systems with Monoliths Enabling Sample Preparation in Proteomics

描述（由申请人提供）：该提案旨在设计、制备和测试含有多孔聚合物整料的微流体装置，该装置将在“样品制备”和复杂混合物中蛋白质的鉴定方面提供无与伦比的性能，并能够充分利用蛋白质组学研究的丰富潜力。它将提供一系列用于处理生物样本的强化工具。由玻璃或塑料制成的装置将包含通过简单的自由基聚合过程从液体前体原位制备的功能性整料。这将提供具有针对所需应用优化的多孔性质的通用大孔整料聚合物，其随后可以通过UV引发的光接枝来官能化。光接枝使得能够将几种不同的功能性（例如疏水性、亲水性、可离子化或亲和性）放置在串联放置或分层或梯度放置的精确位置中，将制备蛋白质的酶促聚集和肽的分离。使用具有精心设计的选择性的分离模块，将困难的蛋白质混合物分离成不太复杂的样品。预计整料的结构性质的优化提供接近100%的样品中存在的所有蛋白质的回收率。还将特别注意设计和控制所有制备工艺，以确保构建模块和复合材料器件的高批次间重现性。将这些设备直接耦合到质谱仪将简化和加速蛋白质组学研究。我们预计，该项目将导致一个全新的阵列的多功能组件和设备的样品制备和质谱研究的蛋白质组或其组件扩展远远超出了目前的最先进的微量分析能力的实际实施。

DESCRIPTION (provided by applicant): This proposal aims at the design, preparation, and testing of microfluidic devices containing porous polymer monoliths that will provide unmatched performance in "sample preparation" and the identification of proteins in complex mixtures and enable full exploitation of the rich potential of proteomic research. It will provide access to an enhanced array of tools for handling biological samples. The devices made of glass or plastic will contain functional monoliths prepared in situ from liquid precursors via a simple free radical polymerization process. This will afford a generic macroporous monolithic polymer with porous properties optimized for the desired application that can subsequently be functionalized by UV intiated photografting. The photografting enables the placement of sevral different functionalities (such as hydrophobic, hydrophilic, ionizable, or affinity) in precise locations placed in series or as layered or gradient chemistriecollection, enzymatic digeston of proteins, and separation of peptides will be prepared. Difficult protein mixtures will be separated into less complex samples using separation modules with carefully designed selectivities. Optimization of structural properties of the monoliths is expected to afford recoveries close to 100% for all of the proteins present in the sample. Specifics attention will also be paid to designing and controlling all preparation processes to ensure high batch-to-batch reproducibility of both building blocks and composite devices. Direct coupling of these devices to a mass spectrometer will simplify and accelerate proteomic research. We expect that this project will result in the practical implementation of a totally new array of versatile components and devices for sample preparation and the mass spectroscopic study of proteomes or their components extending microanalytical capabilities well beyond the current state of the art.