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Quantitative Proteomics Using 18 O Labeling on Target Peptides and Unlabeled Standards
Author(s) -
Truong Hai Ngo Dang,
Lim JaeMin
Publication year - 2021
Publication title -
bulletin of the korean chemical society
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.237
H-Index - 59
ISSN - 1229-5949
DOI - 10.1002/bkcs.12247
Subject(s) - peptide , calibration curve , chemistry , quantitative proteomics , analyte , chromatography , mass spectrometry , label free quantification , calibration , ovalbumin , analytical chemistry (journal) , proteomics , quantitative analysis (chemistry) , detection limit , biochemistry , mathematics , biology , statistics , immune system , immunology , gene
A combination of mass spectrometry (MS) and 18 O‐water labeling has been applied to absolute quantitative proteomics. We introduce a new method called quantitative analysis using unlabeled standards and isotope‐labeled analytes (QAUSIA). An experimental procedure with multiple‐step 18 O labeling is proposed to produce such 18 O‐enriched peptides for investigation of labeling efficiency, response factor, and calibration curve. Furthermore, calibration curves employing unlabeled peptide as internal standard showed good linearity and similar slopes calculated from the mass spectra and the extracted ion chromatograms. Subsequently, the QAUSIA strategy was implemented on the targeted peptide of ovalbumin, and the calibration slope obtained from QAUSIA using the unlabeled synthetic peptide as a standard showed very similar results to the AQUA‐based approach. In addition, the concentration of the QAUSIA strategy using the targeted peptide of ovalbumin was 5.07 ± 0.14 nmol with respect to the theoretical expected value of 5.02 nmol, showing excellent accuracy.