Characterization of Glycosylated Proteins at Subunit Level by HILIC/MS.
Biopharmaceutical proteins
Hydrophilic interaction chromatography
Mass spectrometry
Monoclonal antibodies
N-glycosylation
Protocol
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2021
2021
Historique:
entrez:
28
4
2021
pubmed:
29
4
2021
medline:
24
6
2021
Statut:
ppublish
Résumé
Hydrophilic interaction chromatography (HILIC) coupled to mass spectrometry (MS) is considered as the reference analytical technique for glycans profiling, especially for the characterization of glycosylated protein therapeutics such as monoclonal antibodies (mAbs) and mAbs-related products. Although HILIC/MS is mainly known to profile enzymatically released and fluorescently labeled N-glycans, the recent commercialization of new widepore HILIC amide bonded stationary phases packed with sub-2 μm particles has allowed for remarkable separations also at the subunit level. Here, we describe a simple protocol to perform the mAb glycans profiling at subunit level by HILIC/MS.
Identifiants
pubmed: 33908001
doi: 10.1007/978-1-0716-1241-5_6
doi:
Substances chimiques
Biological Products
0
Rituximab
4F4X42SYQ6
Trastuzumab
P188ANX8CK
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
85-95Références
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