PMID- 29231704
OWN - NLM
STAT- MEDLINE
DCOM- 20190201
LR  - 20231112
IS  - 1520-6882 (Electronic)
IS  - 0003-2700 (Print)
IS  - 0003-2700 (Linking)
VI  - 90
IP  - 2
DP  - 2018 Jan 16
TI  - Site-Specific Glycan Heterogeneity Characterization by Hydrophilic Interaction 
      Liquid Chromatography Solid-Phase Extraction, Reversed-Phase Liquid 
      Chromatography Fractionation, and Capillary Zone Electrophoresis-Electrospray 
      Ionization-Tandem Mass Spectrometry.
PG  - 1223-1233
LID - 10.1021/acs.analchem.7b03912 [doi]
AB  - Reversed-phase chromatographic separation of glycopeptides tends to be dominated 
      by the peptide composition. In contrast, capillary zone electrophoresis 
      separation of glycopeptides is particularly sensitive to the sialic acid 
      composition of the glycan. In this paper, we combine the two techniques to 
      achieve superior N-glycopeptide analysis. Glycopeptides were first isolated from 
      a tryptic digest using hydrophilic interaction liquid chromatography (HILIC) 
      solid-phase extraction. The glycopeptides were separated using reversed-phase 
      ultra high-performance liquid chromatography (UHPLC) to generate four fractions 
      corresponding to different peptide backbones. Capillary zone 
      electrophoresis-electrospray ionization-tandem mass spectrometry (CZE-ESI-MS/MS) 
      was used to analyze the fractions. We applied this method for the analysis of 
      alpha-1-acid glycoprotein (AGP). A total of 268 site-specific N-glycopeptides 
      were detected, representing eight different glycosylation sites from two isomers 
      of AGP. Glycans included tetra-sialic acids with multi N-acetyllactosamine 
      (LacNAc) repeats and unusual pentasialylated terminal sialic acids. 
      Reversed-phase UHPLC coupled with CZE generated  approximately 35% more N-glycopeptides than 
      direct reversed-phase UHPLC-ESI-MS/MS analysis and  approximately 70% more N-glycopeptides than 
      direct CZE-ESI-MS/MS analysis. This approach is a promising tool for global, 
      site-specific glycosylation analysis of highly heterogeneous glycoproteins with 
      mass-limited samples.
FAU - Qu, Yanyan
AU  - Qu Y
AD  - Department of Chemistry and Biochemistry, University of Notre Dame , Notre Dame, 
      Indiana 46556, United States.
FAU - Sun, Liangliang
AU  - Sun L
AUID- ORCID: 0000-0001-8939-5042
AD  - Department of Chemistry, Michigan State University , East Lansing, Michigan 
      48824, United States.
FAU - Zhang, Zhenbin
AU  - Zhang Z
AD  - Department of Chemistry and Biochemistry, University of Notre Dame , Notre Dame, 
      Indiana 46556, United States.
FAU - Dovichi, Norman J
AU  - Dovichi NJ
AUID- ORCID: 0000-0002-1331-3191
AD  - Department of Chemistry and Biochemistry, University of Notre Dame , Notre Dame, 
      Indiana 46556, United States.
LA  - eng
GR  - R01 GM096767/GM/NIGMS NIH HHS/United States
GR  - U54 GM105816/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20180103
PL  - United States
TA  - Anal Chem
JT  - Analytical chemistry
JID - 0370536
RN  - 0 (Glycopeptides)
RN  - 0 (Glycoproteins)
RN  - 0 (Orosomucoid)
RN  - 0 (Polysaccharides)
SB  - IM
MH  - Animals
MH  - Cattle
MH  - Chromatography, High Pressure Liquid/methods
MH  - Chromatography, Reverse-Phase/methods
MH  - Electrophoresis, Capillary/methods
MH  - Glycopeptides/*chemistry
MH  - Glycoproteins/*chemistry
MH  - Glycosylation
MH  - Humans
MH  - Hydrophobic and Hydrophilic Interactions
MH  - Orosomucoid/chemistry
MH  - Polysaccharides/*analysis
MH  - Solid Phase Extraction/methods
MH  - Spectrometry, Mass, Electrospray Ionization/methods
MH  - Tandem Mass Spectrometry/methods
PMC - PMC5771954
MID - NIHMS931570
EDAT- 2017/12/13 06:00
MHDA- 2019/02/02 06:00
PMCR- 2019/01/16
CRDT- 2017/12/13 06:00
PHST- 2017/12/13 06:00 [pubmed]
PHST- 2019/02/02 06:00 [medline]
PHST- 2017/12/13 06:00 [entrez]
PHST- 2019/01/16 00:00 [pmc-release]
AID - 10.1021/acs.analchem.7b03912 [doi]
PST - ppublish
SO  - Anal Chem. 2018 Jan 16;90(2):1223-1233. doi: 10.1021/acs.analchem.7b03912. Epub 
      2018 Jan 3.