Microfluidic Systems with Monoliths Enabling Sample Preparation in Proteomics

描述(申请人提供)：本提案旨在设计、制备和测试含有多孔聚合物整体的微流控装置，这将在“样品制备”和复杂混合物中的蛋白质鉴定方面提供无与伦比的性能，并使蛋白质组学研究的丰富潜力得以充分开发。它将提供一系列增强的处理生物样本的工具。由玻璃或塑料制成的设备将包含由液体前体通过简单的自由基聚合过程原位制备的功能整体。这将提供一种通用型大孔整体式聚合物，具有针对所需应用而优化的多孔性，随后可以通过紫外光引发的光接枝来功能化。光接枝可以将几种不同的功能(如疏水、亲水、可电离或亲和)放置在精确的位置，以串联放置或以分层或梯度化学方式放置，从而制备蛋白质的酶消化和多肽的分离。困难的蛋白质混合物将使用具有精心设计的选择性的分离模块分离成不太复杂的样品。整体结构性质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过程，以确保积木和复合设备的高批次重现性。将这些设备直接连接到质谱仪将简化和加速蛋白质组学研究。我们预计，该项目将导致一系列全新的多功能组件和设备的实际实施，用于样品制备和蛋白质组或其组件的质谱学研究，使微量分析能力远远超出当前技术水平。

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.