Zhang Xumin, Wang Quanhui, Lou Xiaomin, Sun Haidan, Roepstorff Peter, Liu Siqi
Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark.
Methods Mol Biol. 2012;909:17-28. doi: 10.1007/978-1-61779-959-4_2.
2DE coupled with MALDI-MS is one of the most widely used and powerful analytic technologies in proteomics study. The MALDI sample preparation method has been developed and optimized towards the combination of simplicity, sample-cleaning, and sample concentration since its introduction. Here we present a protocol of the so-called Sample loading, Matrix loading, and on-target Wash (SMW) method which fulfills the three criteria by taking advantage of the AnchorChip™ targets. Our method is extremely simple and no pre-desalting or concentration is needed when dealing with samples prepared from 2DE. The protocol is amendable for automation and would pave the road for high-throughput MALDI-MS or MS/MS-based proteomics studies with guaranteed sensitivity and high identification rate. The method has been successfully applied to mouse liver proteome study and so far has been employed in other proteome studies by world-wide researchers.
二维电泳（2DE）与基质辅助激光解吸电离质谱（MALDI-MS）联用是蛋白质组学研究中应用最广泛且功能强大的分析技术之一。自引入以来，MALDI样品制备方法一直朝着简单化、样品净化和样品浓缩相结合的方向发展并得到优化。在此，我们介绍一种所谓的样品加载、基质加载和靶上清洗（SMW）方法的方案，该方法通过利用AnchorChip™靶满足了这三个标准。我们的方法极其简单，处理二维电泳制备的样品时无需预先脱盐或浓缩。该方案适合自动化，将为基于MALDI-MS或串联质谱（MS/MS）的蛋白质组学高通量研究铺平道路，确保灵敏度和高鉴定率。该方法已成功应用于小鼠肝脏蛋白质组研究，迄今为止已被世界各地的研究人员用于其他蛋白质组研究。
