High-speed Analysis of Large Sample Sets - How Can This Key Aspect of the Omics Be Achieved?
High-speed Analysis of Large Sample Sets - How Can This Key Aspect of the Omics Be Achieved?
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DOI：
10.1074/mcp.p120.001997
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发表时间：
2020-11-01
影响因子：
7
通讯作者：
Cramer, Rainer
中科院分区：
文献类型：
--
作者：
Cramer, Rainer
自蛋白质组学开始以来，在单个平台上以每天数千或数百万个样品的速率高速分析大型（蛋白质）组学样品集一直是一个挑战。多年来，基于ESI的MS方法由于其高灵敏度和分析复杂蛋白质组的深度而主导蛋白质组学。然而，尽管在速度上有所改进，但基于ESI的MS方法从根本上受到其样品引入的限制，这排除了离线样品制备/分馏，因为在各个样品/样品级分之间切换所需的时间，因此取决于在线样品制备方法（例如液相色谱法）的速度。基于激光的电离方法具有从一个样品移动到下一个样品而没有这些限制的优点，主要受现代样品台的速度限制，即样品之间的10 ms或更短。该速度与现代高性能质谱仪的数据采集速度相匹配，而激光器的脉冲重复率（> 1 kHz）提供了足够数量的解吸/电离事件，用于在样品台的上述速度下从每个样品成功检测离子信号。基于激光的电离方法的其他优点包括与ESI MS相比，对样品添加剂和污染的耐受性通常更高，以及用于解吸的激光的非接触和脉冲性质，从而降低了交叉污染的风险。此外，MALDI的新发展扩大了其分析能力，现在能够充分利用高性能混合质量分析仪及其在灵敏度和MS/MS分析方面的优势，产生类似ESI的多电荷分析物离子的稳定产量。因此，这些新的发展和基于激光的方法固有的高速度现在为解决组学中的极端样品分析速度提供了良好的基础。
High-speed analysis of large (prote)omics sample sets at the rate of thousands or millions of samples per day on a single platform has been a challenge since the beginning of proteomics. For many years, ESI-based MS methods have dominated proteomics because of their high sensitivity and great depth in analyzing complex proteomes. However, despite improvements in speed, ESI-based MS methods are fundamentally limited by their sample introduction, which excludes off-line sample preparation/fractionation because of the time required to switch between individual samples/sample fractions, and therefore being dependent on the speed of on-line sample preparation methods such as liquid chromatography. Laser-based ionization methods have the advantage of moving from one sample to the next without these limitations, being mainly restricted by the speed of modern sample stages, i.e. 10 ms or less between samples. This speed matches the data acquisition speed of modern high-performing mass spectrometers whereas the pulse repetition rate of the lasers (>1kHz) provides a sufficient number of desorption/ionization events for successful ion signal detection from each sample at the above speed of the sample stages. Other advantages of laser-based ionization methods include the generally higher tolerance to sample additives and contamination compared with ESI MS, and the contact-less and pulsed nature of the laser used for desorption, reducing the risk of cross-contamination. Furthermore, new developments in MALDI have expanded its analytical capabilities, now being able to fully exploit high-performing hybrid mass analyzers and their strengths in sensitivity and MS/MS analysis by generating an ESI-like stable yield of multiply charged analyte ions. Thus, these new developments and the intrinsically high speed of laser-based methods now provide a good basis for tackling extreme sample analysis speed in the omics.