2019
DOI: 10.1021/acs.analchem.9b01983
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Optimized Fragmentation for Quantitative Analysis of Fucosylated N-Glycoproteins by LC-MS-MRM

Abstract: Quantitative analysis of site specific glycoforms of proteins is technically challenging but highly desirable; resolution of the fucosylated glycoforms is of particular interest due to their biological importance. In this study, we developed a sensitive and specific LC-MS-MRM quantification method that distinguishes the outer arm and core fucosylated configurations of the Nglycopeptides. We take advantage of limited fragmentation of the glycopeptides at low collision energy CID to produce linkage-specific Y-io… Show more

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Cited by 16 publications
(18 citation statements)
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“…The limitation of this assay is label-free quantitation, while complimentary approaches utilizing isobaric mass tags 50 or metabolic 15 N/ 13 C labeling strategy 51 can improve the precision of relative quantitation of N-glycopeptides. Absolute quantitation using multiple reaction monitoring (MRM) 52,53 is a future direction, although it is challenging due to the complexity of glycopeptides composed by glycan and peptide motifs. Future studies on a larger cohort of NASH patients will be needed to validate our findings, where an MRM-based assay can be employed for absolute quantitation of the N-glycopeptide marker candidates in NASH patients.…”
Section: Performance Of Hp N-glycopeptide Markers In Afp-negative Pat...mentioning
confidence: 99%
“…The limitation of this assay is label-free quantitation, while complimentary approaches utilizing isobaric mass tags 50 or metabolic 15 N/ 13 C labeling strategy 51 can improve the precision of relative quantitation of N-glycopeptides. Absolute quantitation using multiple reaction monitoring (MRM) 52,53 is a future direction, although it is challenging due to the complexity of glycopeptides composed by glycan and peptide motifs. Future studies on a larger cohort of NASH patients will be needed to validate our findings, where an MRM-based assay can be employed for absolute quantitation of the N-glycopeptide marker candidates in NASH patients.…”
Section: Performance Of Hp N-glycopeptide Markers In Afp-negative Pat...mentioning
confidence: 99%
“…Then only the selected specific precursor ion is subjected to collision‐induction to remove the interference of other product ions, and finally only the selected specific product ion is collected for mass spectrometry signals [33]. Because the Triple Quadrupole System (TQS) is a mass spectrometry system sensitive to single mass‐to‐charge ratio scanning, it is a mass spectrometer suitable for MRM analysis [34,35].…”
Section: Quantification Of Anthocyaninsmentioning
confidence: 99%
“…A more serious concern, on the other hand, is the facile Fuc ion migration and rearrangement behavior during the analysis of soft ionization MS/MS that unavoidably caused ambiguous even wrong results, despite chemical derivatization with easily ionized groups could overcome the issue of relative low ionization efficiency of Fuc ( Harvey et al., 2002 ; Nwosu et al., 2015 ). Chemical modifications ( Lattova et al., 2019 ) and fine controlling of the CID energy ( Acs et al., 2018 ; Yuan et al., 2019b ), for example, could minimize the consecutive processes of the facile Fuc ion migration and neutral Fuc residue loss during the soft ionization MS/MS enabling the discrimination of core and antenna fucosylation. But the sensitivity was reluctantly compromised, and still there were several (the core fucosylation) to a dozen (the antenna fucosylation) percentage of interferences, merely offering their relative abundance ratios if reference standard compounds were not available.…”
Section: Introductionmentioning
confidence: 99%