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Relative and Absolute Quantitation in Mass Spectrometry–Based Proteomics
Annual Review of Analytical Chemistry ( IF 8 ) Pub Date : 2016-06-24 00:00:00 , DOI: 10.1146/annurev-anchem-061516-045357
J. Astor Ankney 1 , Adil Muneer 1 , Xian Chen 1, 2
Affiliation  

Mass spectrometry–based quantitative proteomics is a powerful tool for gaining insights into function and dynamics of biological systems. However, peptides with different sequences have different ionization efficiencies, and their intensities in a mass spectrum are not correlated with their abundances. Therefore, various label-free or stable isotope label–based quantitation methods have emerged to assist mass spectrometry to perform comparative proteomic experiments, thus enabling nonbiased identification of thousands of proteins differentially expressed in healthy versus diseased cells. Here, we discuss the most widely used label-free and metabolic-, enzymatic-, and chemical labeling–based proteomic strategies for relative and absolute quantitation. We summarize the specific strengths and weaknesses of each technique in terms of quantification accuracy, proteome coverage, multiplexing capability, and robustness. Applications of each strategy for solving specific biological complexities are also presented.

中文翻译:


基于质谱的蛋白质组学中的相对和绝对定量

基于质谱的定量蛋白质组学是了解生物系统功能和动力学的有力工具。然而,具有不同序列的肽具有不同的电离效率,并且它们在质谱中的强度与它们的丰度不相关。因此,出现了各种基于无标记或稳定同位素标记的定量方法,以协助质谱法进行比较蛋白质组学实验,从而能够无偏鉴定健康细胞和患病细胞中数千种差异表达的蛋白质。在这里,我们讨论了最广泛使用的无标记和基于代谢,酶促和化学标记的蛋白质组学策略,用于相对和绝对定量。我们总结了每种技术在定量准确性,蛋白质组覆盖率,多路复用能力和鲁棒性方面的优势和劣势。还介绍了解决特定生物学复杂性的每种策略的应用。

更新日期:2016-06-24
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