Microfluidic Systems with Monoliths Enabling Sample Preparation in Proteomics

描述（由申请人提供）：本提案旨在设计，制备和测试含有多孔聚合物单体的微流控装置，该装置将在“样品制备”和复杂混合物中蛋白质的鉴定中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。它将为处理生物样品提供一系列增强的工具。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单体。这将提供一种通用的大孔整体聚合物，其多孔性能针对所需的应用进行了优化，随后可以通过UV引发的光接枝实现功能化。光接枝可以将几种不同的功能（如疏水、亲水性、可电离或亲和性）放置在精确的位置上，以串联或分层或梯度的方式进行化学收集，酶促蛋白质消化和多肽分离。困难的蛋白质混合物将分离成较不复杂的样品使用分离模块与精心设计的选择性。单体结构性能的优化有望为样品中存在的所有蛋白质提供接近100%的回收率。还将特别注意设计和控制所有制备工艺，以确保构建块和复合设备的高批对批可重复性。这些设备与质谱仪的直接耦合将简化和加速蛋白质组学研究。我们期望这个项目将导致一种全新的通用组件和设备阵列的实际实施，用于样品制备和蛋白质组或其组件的质谱研究，将微观分析能力扩展到远远超出当前技术水平的水平。

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.