Luk Vivienne N, Wheeler Aaron R
Department of Chemistry, University of Toronto, 80 Street George St., Toronto, Ontario M5S 3H6, Canada.
Anal Chem. 2009 Jun 1;81(11):4524-30. doi: 10.1021/ac900522a.
A common characteristic for proteomic analyses is the need for extensive biochemical processing. Digital microfluidics (DMF), a technique characterized by the manipulation of discrete microdroplets (100 nL-10 microL) on an open array of electrodes, is a good match for carrying out rapid, automated solution-phase reactions. Here, we report a DMF-based method integrating several common processing steps in proteomics, including reduction, alkylation, and enzymatic digestion. Fluorogenic assays were used to quantitatively evaluate the kinetics and reproducibility of each reaction step, and MALDI-MS was used for qualitative confirmation. The method is fast, facile, and reproducible, and thus has the potential to be a useful new tool in proteomics.
蛋白质组学分析的一个共同特点是需要进行广泛的生化处理。数字微流控（DMF）是一种通过在开放电极阵列上操纵离散微滴（100 nL - 10 μL）为特征的技术，非常适合进行快速、自动化的溶液相反应。在此，我们报告了一种基于DMF的方法，该方法整合了蛋白质组学中的几个常见处理步骤，包括还原、烷基化和酶解。使用荧光测定法定量评估每个反应步骤的动力学和重现性，并使用基质辅助激光解吸电离质谱（MALDI-MS）进行定性确认。该方法快速、简便且可重现，因此有潜力成为蛋白质组学中一种有用的新工具。
