Sample preparation is a crucial step in bottom-up proteomics. Analytical performances of bottom-up proteomics can be improved by the miniaturization of sample preparation. Many microfluidic devices have been designed in the field of proteomics, but many of them are not capable of handling complex samples and do not integrate the processing and digestion steps. We propose a ChipFilter Proteolysis (CFP) microfluidic device as a proteomics reactor for the miniaturization of protein sample processing and digestion steps, whose design is closely related to the experimental setup of filter-aided sample processing, even if no denaturing surfactant is required. The microchip has two reaction chambers of 0.6 μL volume separated by a protein filtration membrane in regenerated cellulose (10kD cutoff) that will concentrate or retain large polypeptides and will release small molecules. Cell lysis, protein concentration, and rapid chemical or enzymatic treatment can be performed in the ChipFilter. Complex proteomic samples like yeast protein extract or whole human cells proteome have been successfully analyzed with our microchip. Compared with the membrane-based commercial ultracentrifugation cartridge, our microfluidic device offered a better proteome coverage with 10 times less starting material and 8 times faster protocol duration.

中文翻译：

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使用与 NanoLC-MS/MS 耦合的芯片过滤器进行芯片上样品制备，用于自下而上的蛋白质组学。

样品制备是自下而上蛋白质组学的关键步骤。样品制备的小型化可以提高自下而上蛋白质组学的分析性能。蛋白质组学领域已经设计了许多微流体装置，但其中许多不能处理复杂的样品，也没有整合处理和消化步骤。我们提出了一种 ChipFilter Proteolysis (CFP) 微流体装置作为蛋白质组学反应器，用于蛋白质样品处理和消化步骤的小型化，其设计与过滤辅助样品处理的实验设置密切相关，即使不需要变性表面活性剂。微芯片有两个 0 的反应室。6 μL 体积由再生纤维素（10kD 截断值）中的蛋白质过滤膜分离，可浓缩或保留大多肽并释放小分子。细胞裂解、蛋白质浓缩和快速化学或酶处理可以在 ChipFilter 中进行。我们的微芯片已成功分析了复杂的蛋白质组学样品，如酵母蛋白提取物或全人类细胞蛋白质组。与基于膜的商业超速离心柱相比，我们的微流体装置提供了更好的蛋白质组覆盖率，起始材料减少了 10 倍，方案持续时间缩短了 8 倍。我们的微芯片已成功分析了复杂的蛋白质组学样品，如酵母蛋白提取物或全人类细胞蛋白质组。与基于膜的商业超速离心柱相比，我们的微流体装置提供了更好的蛋白质组覆盖率，起始材料减少了 10 倍，方案持续时间缩短了 8 倍。我们的微芯片已成功分析了复杂的蛋白质组学样品，如酵母蛋白提取物或全人类细胞蛋白质组。与基于膜的商业超速离心柱相比，我们的微流体装置提供了更好的蛋白质组覆盖率，起始材料减少了 10 倍，方案持续时间缩短了 8 倍。