Chapter title |
Absolute Quantification of Middle- to High-Abundant Plasma Proteins via Targeted Proteomics
|
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Chapter number | 29 |
Book title |
Serum/Plasma Proteomics
|
Published in |
Methods in molecular biology, July 2017
|
DOI | 10.1007/978-1-4939-7057-5_29 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7056-8, 978-1-4939-7057-5
|
Authors |
Julia Dittrich, Uta Ceglarek |
Editors |
David W. Greening, Richard J. Simpson |
Abstract |
The increasing number of peptide and protein biomarker candidates requires expeditious and reliable quantification strategies. The utilization of liquid chromatography coupled to quadrupole tandem mass spectrometry (LC-MS/MS) for the absolute quantitation of plasma proteins and peptides facilitates the multiplexed verification of tens to hundreds of biomarkers from smallest sample quantities. Targeted proteomics assays derived from bottom-up proteomics principles rely on the identification and analysis of proteotypic peptides formed in an enzymatic digestion of the target protein. This protocol proposes a procedure for the establishment of a targeted absolute quantitation method for middle- to high-abundant plasma proteins waiving depletion or enrichment steps. Essential topics as proteotypic peptide identification and LC-MS/MS method development as well as sample preparation and calibration strategies are described in detail. |
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