Novel method to specifically determine the structures of non-N297 glycans in IgGs.
IgG glycosylation
antibody glycosylation
endoglycosidase S2
glycan analytics
Journal
Glycobiology
ISSN: 1460-2423
Titre abrégé: Glycobiology
Pays: England
ID NLM: 9104124
Informations de publication
Date de publication:
03 06 2023
03 06 2023
Historique:
received:
22
07
2022
revised:
22
02
2023
accepted:
24
02
2023
medline:
8
6
2023
pubmed:
8
3
2023
entrez:
7
3
2023
Statut:
ppublish
Résumé
Monoclonal antibodies comprise a major class of biologic therapeutics and are also extensively studied in immunology. Given the importance of glycans on antibodies, fluorescent labeling of enzymatically released glycans and their LC/MS analysis is routinely used for in-depth characterization of antibody glycosylation. In this technical note, we propose a method for facile characterization of glycans in the variable region of antibodies using sequential enzymatic digests with Endoglycosidase-S2 and RapidTM Peptide-N-Glycosidase-F followed by labeling with a fluorescent dye carrying an NHS-carbamate moiety. The results and proposed mechanism also suggest that the choice of glycosidases along with the labeling chemistry is critical for accurate glycan analysis for a desired application.
Identifiants
pubmed: 36881660
pii: 7070571
doi: 10.1093/glycob/cwad020
doi:
Substances chimiques
Polysaccharides
0
Immunoglobulin G
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
364-368Informations de copyright
© The Author(s) 2023. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.