TY  - JOUR
A1  - Fang, Pan
A1  - Ji, Yanlong
A1  - Silbern, Ivan
A1  - Viner, Rosa
A1  - Oellerich, Thomas
A1  - Pan, Kuan-Ting
A1  - Urlaub, Henning
T1  - Evaluation and optimization of high-field asymmetric waveform ion-mobility spectrometry for multiplexed quantitative site-specific N-glycoproteomics
T2  - Analytical chemistry
N2  - The heterogeneity and complexity of glycosylation hinder the depth of site-specific glycoproteomics analysis. High-field asymmetric-waveform ion-mobility spectrometry (FAIMS) has been shown to improve the scope of bottom-up proteomics. The benefits of FAIMS for quantitative N-glycoproteomics have not been investigated yet. In this work, we optimized FAIMS settings for N-glycopeptide identification, with or without the tandem mass tag (TMT) label. The optimized FAIMS approach significantly increased the identification of site-specific N-glycopeptides derived from the purified immunoglobulin M (IgM) protein or human lymphoma cells. We explored in detail the changes in FAIMS mobility caused by N-glycopeptides with different characteristics, including TMT labeling, charge state, glycan type, peptide sequence, glycan size, and precursor m/z. Importantly, FAIMS also improved multiplexed N-glycopeptide quantification, both with the standard MS2 acquisition method and with our recently developed Glyco-SPS-MS3 method. The combination of FAIMS and Glyco-SPS-MS3 methods provided the highest quantitative accuracy and precision. Our results demonstrate the advantages of FAIMS for improved mass spectrometry-based qualitative and quantitative N-glycoproteomics.
KW  - Carbohydrates
KW  - Chemical biology
KW  - Mass spectrometry
KW  - Mathematical methods
KW  - Peptides and proteins
Y1  - 2021
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/62958
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-629589
SN  - 1520-6882
VL  - 93
IS  - 25
SP  - 8846
EP  - 8855
PB  - American Chemical Society
CY  - Columbus, Ohio
ER  -