Leiden University Medical Center (LUMC), Center for Proteomics and Metabolomics, P.O. Box 9600, 2300 RC Leiden, Netherlands.
J Anal Methods Chem. 2015;2015:250131. doi: 10.1155/2015/250131. Epub 2015 Jan 27.
For large-scale and standardized applications in mass spectrometry- (MS-) based proteomics automation of each step is essential. Here we present high-throughput sample preparation solutions for balancing the speed of current MS-acquisitions and the time needed for analytical workup of body fluids. The discussed workflows reduce body fluid sample complexity and apply for both bottom-up proteomics experiments and top-down protein characterization approaches. Various sample preparation methods that involve solid-phase extraction (SPE) including affinity enrichment strategies have been automated. Obtained peptide and protein fractions can be mass analyzed by direct infusion into an electrospray ionization (ESI) source or by means of matrix-assisted laser desorption ionization (MALDI) without further need of time-consuming liquid chromatography (LC) separations.
对于基于质谱（MS）的蛋白质组学的大规模和标准化应用，每个步骤的自动化都是至关重要的。在这里，我们提出了高通量样品制备解决方案，以平衡当前 MS 采集的速度和体液分析工作所需的时间。所讨论的工作流程降低了体液样品的复杂性，适用于从头蛋白质组学实验和自上而下的蛋白质特征分析方法。各种涉及固相萃取（SPE）的样品制备方法，包括亲和富集策略，已经实现了自动化。获得的肽和蛋白质馏分可以通过直接注入电喷雾电离（ESI）源进行质量分析，或者通过基质辅助激光解吸电离（MALDI）进行分析，而无需耗时的液相色谱（LC）分离。
