Dataset Information

Hydrogen deuterium exchange epitope mapping of glycosylated epitopes enabled by online immobilized glycosidase

ABSTRACT: Numerous mAbs are known to recognize N-glycosylated epitopes or to bind in close proximity to an N-glycan site, however, glycosylated protein sites are typically obscured from HDX detection as a result of the inherent heterogeneity of glycans. To overcome this limitation, we covalently immobilized the glycosidase PNGase Dj on solid resin and incorporated it into an online HDX-MS workflow for post-HDX deglycosylation. The resin-immobilized PNGase Dj exhibits a robust tolerance to various buffer conditions, and is employed in a column format that can be readily adapted into a typical HDX-MS platform. Using this system, we were able to obtain full sequence coverage of the SARS-CoV-2 receptor binding domain (RBD) and to map the glycosylated epitope of the glycan-binding mAb S309 to the RBD.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Timothy O'Leary

LAB HEAD: Patrick R. Griffin

PROVIDER: PXD041204 | Pride | 2024-05-24

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	AGP_sequence_used_for_HDX_GlycN_to_D.fasta	Fasta
	Fetuin_A_bovine_sequence.fasta	Fasta
	HRP_sequence.fasta	Fasta
	RBD_sequence_used_for_HDX_GlycN_to_D.fasta	Fasta
	TO_122_2columns_pepsin1_pngaseDJ2_2uM_AGP_Naphos_50TCEP.csv	Csv

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Publications

Hydrogen-Deuterium Exchange Epitope Mapping of Glycosylated Epitopes Enabled by Online Immobilized Glycosidase.

O'Leary Timothy R TR Balasubramaniam Deepa D Hughes Kristin K Foster Denisa D Boyles Jeffrey J Coleman Kristina K Griffin Patrick R PR

Analytical chemistry 20230628 27

Hydrogen-deuterium exchange coupled with mass spectrometry (HDX-MS) is widely used for monoclonal antibody (mAb) epitope mapping, which aids in the development of therapeutic mAbs and vaccines, as well as enables the understanding of viral immune evasion. Numerous mAbs are known to recognize N-glycosylated epitopes and to bind in close proximity to an <i>N</i>-glycan site; however, glycosylated protein sites are typically obscured from HDX detection as a result of the inherent heterogeneity of g ...[more]

PMID: 37379434

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