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PSAQ™ standards for accurate MS–based quantification of proteins: from the concept to biomedical applications
Author(s) -
Picard Guillaume,
Lebert Dorothée,
Louwagie Mathilde,
Adrait Annie,
Huillet Céline,
Vandenesch François,
Bruley Christophe,
Garin Jérôme,
Jaquinod Michel,
Brun Virginie
Publication year - 2012
Publication title -
journal of mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 1076-5174
DOI - 10.1002/jms.3106
Subject(s) - chemistry , sample preparation , mass spectrometry , quantitative proteomics , computational biology , label free quantification , sample (material) , chromatography , biochemical engineering , nanotechnology , proteomics , biochemistry , materials science , biology , engineering , gene
Absolute protein quantification, i.e. determining protein concentrations in biological samples, is essential to our understanding of biological and physiopathological phenomena. Protein quantification methods based on the use of antibodies are very effective and widely used. However, over the last ten years, absolute protein quantification by mass spectrometry has attracted considerable interest, particularly for the study of systems biology and as part of biomarker development. This interest is mainly linked to the high multiplexing capacity of MS analysis, and to the availability of stable‐isotope‐labelled standards for quantification. This article describes the details of how to produce, control the quality and use a specific type of standard: Protein Standard Absolute Quantification (PSAQ™) standards. These standards are whole isotopically labelled proteins, analogues of the proteins to be assayed. PSAQ standards can be added early during sample treatment, thus they can correct for protein losses during sample prefractionation and for incomplete sample digestion. Because of this, quantification of target proteins is very accurate and precise using these standards. To illustrate the advantages of the PSAQ method, and to contribute to the increase in its use, selected applications in the biomedical field are detailed here. Copyright © 2012 John Wiley & Sons, Ltd.