2015
DOI: 10.1038/nprot.2015.015
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Building high-quality assay libraries for targeted analysis of SWATH MS data

Abstract: Targeted proteomics by selected/multiple reaction monitoring (S/MRM) or, on a larger scale, by SWATH (sequential window acquisition of all theoretical spectra) MS (mass spectrometry) typically relies on spectral reference libraries for peptide identification. Quality and coverage of these libraries are therefore of crucial importance for the performance of the methods. Here we present a detailed protocol that has been successfully used to build high-quality, extensive reference libraries supporting targeted pr… Show more

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Cited by 296 publications
(314 citation statements)
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“…However, it is important to quantify CML-and CELmodified albumin, as these modifications constitute the predominant AGEs. Several mass-spectrometry-based targeted quantification approaches, including multiple reaction monitoring, parallel reaction monitoring, and SWATH, have become powerful tools and rely heavily on fragment ion library (39,40). In this context, we constructed a fragment ion library for the synthetically AML-, CML-, and CEL-modified peptides of albumin by using high-resolution accurate mass spectrometer followed by rigorous inspection and validation of MS/MS spectra.…”
Section: Discussionmentioning
confidence: 99%
“…However, it is important to quantify CML-and CELmodified albumin, as these modifications constitute the predominant AGEs. Several mass-spectrometry-based targeted quantification approaches, including multiple reaction monitoring, parallel reaction monitoring, and SWATH, have become powerful tools and rely heavily on fragment ion library (39,40). In this context, we constructed a fragment ion library for the synthetically AML-, CML-, and CEL-modified peptides of albumin by using high-resolution accurate mass spectrometer followed by rigorous inspection and validation of MS/MS spectra.…”
Section: Discussionmentioning
confidence: 99%
“…To extract quantitative information from such digital SWATH-MS data, high-quality assay libraries are required (109,111). Such libraries contain retention-time and fragmentation information of the peptides to be quantified.…”
Section: Identification and Quantification Of Mhc-associated Peptidesmentioning
confidence: 99%
“…Recent advances in proteomics make it possible to quantify many proteins in large numbers of samples in model organisms with small genomes (Picotti et al, 2013;Schubert et al, 2015). However, these technologies are not yet applicable to plants.…”
Section: Introductionmentioning
confidence: 99%