Mass spectrometry-based quantification

被引:25
作者
DeSouza, Leroi V.
Siu, K. W. Michael [1 ]
机构
[1] York Univ, Dept Chem, Toronto, ON M3J 1P3, Canada
关键词
Relative quantification; LC-MS/MS; LC-MRM; iTRAQ; ICAT; SILAC; Proteolytic cleavage; Stable isotope labeling; Mass tags; COMPLEX PROTEIN MIXTURES; CELL-CULTURE SILAC; QUANTITATIVE PROTEOMICS; AMINO-ACIDS; ABSOLUTE QUANTIFICATION; GEL-ELECTROPHORESIS; AFFINITY TAGS; EXPRESSION; O-18; IDENTIFICATION;
D O I
10.1016/j.clinbiochem.2012.10.025
中图分类号
R446 [实验室诊断]; R-33 [实验医学、医学实验];
学科分类号
1001 ;
摘要
Objective: This review describes the most common methods currently used for mass spectrometry-based quantification in proteomic applications. Design and methods: Quantification can be performed using either labeled or unlabeled approaches. Labeled approaches rely on mass-tagging of peptide reference standards either by direct synthesis or through chemical or metabolic means. Following labeling, samples are pooled and analyzed as one, thereby circumventing any potential error that could result from run-to-run variability. Unlabeled approaches rely on run-to-run comparisons between test and reference samples and assume a highly reproducible separation and similar sample compositions. Results: A number of commercial labeling reagents are now available and each of these are described along with their strengths and limitations. Conclusions: The choice of method will ultimately depend, not just on financial considerations, but also on the context of the experiment. (C) 2012 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.
引用
收藏
页码:421 / 431
页数:11
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