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Quantitative measurements of N ‐linked glycoproteins in human plasma by SWATH ‐ MS
Author(s) -
Liu Yansheng,
Hüttenhain Ruth,
Surinova Silvia,
Gillet Ludovic CJ,
Mouritsen Jeppe,
Brunner Roland,
Navarro Pedro,
Aebersold Ruedi
Publication year - 2013
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201200417
Subject(s) - reproducibility , biomarker , glycoprotein , biomarker discovery , chromatography , human plasma , chemistry , peptide , analyte , sample preparation , analytical chemistry (journal) , proteomics , biochemistry , gene
SWATH ‐ MS is a data‐independent acquisition method that generates, in a single measurement, a complete recording of the fragment ion spectra of all the analytes in a biological sample for which the precursor ions are within a predetermined m/z versus retention time window. To assess the performance and suitability of SWATH ‐ MS ‐based protein quantification for clinical use, we compared SWATH ‐ MS and SRM ‐ MS ‐based quantification of N ‐linked glycoproteins in human plasma, a commonly used sample for biomarker discovery. Using dilution series of isotopically labeled heavy peptides representing biomarker candidates, the LOQ of SWATH ‐ MS was determined to reach 0.0456 fmol at peptide level by targeted data analysis, which corresponds to a concentration of 5–10 ng protein/mL in plasma, while SRM reached a peptide LOQ of 0.0152 fmol. Moreover, the quantification of endogenous glycoproteins using SWATH ‐ MS showed a high degree of reproducibility, with the mean CV of 14.90%, correlating well with SRM results ( R 2 = 0.9784). Overall, SWATH ‐ MS measurements showed a slightly lower sensitivity and a comparable reproducibility to state‐of‐the‐art SRM measurements for targeted quantification of the N ‐glycosites in human blood. However, a significantly larger number of peptides can be quantified per analysis. We suggest that SWATH ‐ MS analysis combined with N ‐glycoproteome enrichment in plasma samples is a promising integrative proteomic approach for biomarker discovery and verification.