Bantscheff Marcus, Schirle Markus, Sweetman Gavain, Rick Jens, Kuster Bernhard
Cellzome AG, Meyerhofstrasse 1, 69254, Heidelberg, Germany.
Anal Bioanal Chem. 2007 Oct;389(4):1017-31. doi: 10.1007/s00216-007-1486-6. Epub 2007 Aug 1.
The quantification of differences between two or more physiological states of a biological system is among the most important but also most challenging technical tasks in proteomics. In addition to the classical methods of differential protein gel or blot staining by dyes and fluorophores, mass-spectrometry-based quantification methods have gained increasing popularity over the past five years. Most of these methods employ differential stable isotope labeling to create a specific mass tag that can be recognized by a mass spectrometer and at the same time provide the basis for quantification. These mass tags can be introduced into proteins or peptides (i) metabolically, (ii) by chemical means, (iii) enzymatically, or (iv) provided by spiked synthetic peptide standards. In contrast, label-free quantification approaches aim to correlate the mass spectrometric signal of intact proteolytic peptides or the number of peptide sequencing events with the relative or absolute protein quantity directly. In this review, we critically examine the more commonly used quantitative mass spectrometry methods for their individual merits and discuss challenges in arriving at meaningful interpretations of quantitative proteomic data.
对生物系统两种或更多生理状态之间的差异进行定量分析，是蛋白质组学中最重要但也最具挑战性的技术任务之一。除了通过染料和荧光团进行差异蛋白质凝胶或印迹染色的经典方法外，基于质谱的定量方法在过去五年中越来越受欢迎。这些方法大多采用差异稳定同位素标记来创建一个特定的质量标签，该标签可被质谱仪识别，同时为定量分析提供基础。这些质量标签可以通过以下方式引入蛋白质或肽中：（i）代谢方式，（ii）化学方法，（iii）酶促方法，或（iv）通过添加合成肽标准品提供。相比之下，无标记定量方法旨在将完整蛋白水解肽的质谱信号或肽测序事件的数量直接与相对或绝对蛋白质数量相关联。在本综述中，我们批判性地审视了更常用的定量质谱方法的各自优点，并讨论了在对定量蛋白质组学数据进行有意义的解释时所面临的挑战。
