Anand Sushma, Samuel Monisha, Ang Ching-Seng, Keerthikumar Shivakumar, Mathivanan Suresh
Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, VIC, 3086, Australia.
Department of Physiology, Anatomy and Microbiology, School of Life Sciences, La Trobe University, Bundoora, Melbourne, VIC, 3086, Australia.
Methods Mol Biol. 2017;1549:31-43. doi: 10.1007/978-1-4939-6740-7_4.
The precise quantification of changes between various physiological states in a biological system is highly complex in nature. Over the past few years, in combination with classical methods, mass spectrometry based approaches have become an indispensable tool in deciphering exact abundance of proteins in composite mixtures. The technique is now well established and employs both label-based and label-free quantitation strategies. Label-based quantitation methods utilize stable isotope labels which are incorporated within the peptides, introducing an expectable mass difference within the two or more experimental conditions. In contrast, label-free proteomics quantitates both relative and absolute protein quantity by utilizing signal intensity and spectral counting of peptides. This chapter focuses on the commonly used quantitative mass spectrometry methods for high-throughput proteomic analysis.
生物系统中各种生理状态之间变化的精确量化本质上极为复杂。在过去几年里,基于质谱的方法与经典方法相结合,已成为解析复合混合物中蛋白质确切丰度的不可或缺的工具。该技术现已成熟,并采用基于标记和无标记定量策略。基于标记的定量方法利用稳定同位素标记,将其掺入肽段中,在两个或更多实验条件下引入可预期的质量差异。相比之下,无标记蛋白质组学通过利用肽段的信号强度和谱图计数来定量相对和绝对蛋白质数量。本章重点介绍用于高通量蛋白质组分析的常用定量质谱方法。