Journal
ANALYTICAL BIOCHEMISTRY
Volume 417, Issue 1, Pages 80-88Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2011.05.028
Keywords
Hydrophilic interaction chromatography; Mass spectrometry; Glycoprotein; Characterization; Glycopeptides; Glycans
Funding
- China National Key Basic Research Program [2011CB910603]
- National Natural Science Foundation of China [20735005]
Ask authors/readers for more resources
A new hydrophilic interaction chromatography (HILIC) column packed with amide 1.7 mu m sorbent was applied to the characterization of glycoprotein digests. Due to the impact of the hydrophilic carbohydrate moiety, glycopeptides were more strongly retained on the column and separated from the remaining nonglycosylated peptides present in the digest. The glycoforms of the same parent peptide were also chromatographically resolved and analyzed using ultraviolet and mass spectrometry detectors. The HILIC method was applied to glyco-profiling of a therapeutic monoclonal antibody and proteins with several N-linked and O-linked glycosylation sites. For characterization of complex proteins with multiple glycosylation sites we utilized 2D LC, where RP separation dimension was used for isolation of glycopeptides and HILIC for resolution of peptide glycoforms. The analysis of site-specific glycan microheterogeneity was illustrated for the CD44 fusion protein. (C) 2011 Elsevier Inc. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available