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- Title
Quantitative Proteomics Using <sup>18</sup>O Labeling on Target Peptides and Unlabeled Standards.
- Authors
Truong, Hai Ngo Dang; Lim, Jae‐Min
- Abstract
A combination of mass spectrometry (MS) and 18O‐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 18O labeling is proposed to produce such 18O‐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.
- Subjects
PROTEOMICS; MASS spectrometry; PEPTIDES; IONS spectra; EXPECTED returns; DEAMINATION; DRUG labeling
- Publication
Bulletin of the Korean Chemical Society, 2021, Vol 42, Issue 4, p633
- ISSN
0253-2964
- Publication type
Article
- DOI
10.1002/bkcs.12247