PXD062588

PXD062588 is an original dataset announced via ProteomeXchange.

Title	Deciphering Minor Glycan Signatures: Cross-Serotype Recombinant Adeno-Associated Virus (rAAV) Glycan Profiling Using Multimodal Glycoproteomics Technologies
Description	Glycan analysis of viral vectors is essential for pharmaceutical quality evaluation. Recent advances in mass spectrometry (MS)-based analytical technologies have identified glycosylation in adeno-associated viruses (AAVs). However, reported glycan structures vary between studies, due to the extremely low abundance (<1%) of glycosylated particles and the presence of hitchhiker molecules, making AAV glycosylation a topic of debate. This underscores the need for standardized and highly sensitive detection methods. Here, we present a multimodal glycoproteomic approach to characterize glycosylation across six serotypes of 28 recombinant AAVs (rAAVs). An ultrasensitive lectin microarray, using a broad-reactive anti-AAV antibody, detected O-glycan-binding lectin signals in rAAV6, consistent with previous findings. Notably, a unique binding signal was observed with Urtica dioica lectin (UDA). Liquid chromatography-tandem mass spectrometry (LC-MS/MS), combined with UDA-based glycoprotein/glycopeptide dual enrichment and glycopeptide identification, confirmed the predominant presence of a divalent high-mannose N-glycosylated peptide from a hitchhiking host-derived glycoprotein, the AAV receptor. In contrast, monovalent high-mannose N-glycosylated peptides were identified using a single-step protein-level enrichment approach, suggesting an avidity effect in UDA binding. These findings highlight the significance of advanced multimodal glycoproteomic in rAAV characterization and offer new insights into host-derived hitchhiker glycoproteins present in AAV preparations.
HostingRepository	jPOST
AnnounceDate	2026-04-04
AnnouncementXML	Submission_2026-04-03_08:00:04.826.xml
DigitalObjectIdentifier
ReviewLevel	Non peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Hiroaki Sakaue
SpeciesList	scientific name: Cloning vector AAV; NCBI TaxID: 1116368;
ModificationList	L-cysteine methyl disulfide; Ammonia-loss; 2-pyrrolidone-5-carboxylic acid (Gln); L-methionine sulfoxide
Instrument	timsTOF fleX MALDI-2

Revision	Datetime	Status	ChangeLog Entry
0	2025-04-03 23:21:21	ID requested
⏵ 1	2026-04-03 08:00:05	announced

Dataset with its publication pending

submitter keyword: AAV, glycosylation, glycoproteomics

Atsushi Kuno
lab head
Hiroaki Sakaue
contact affiliation	National Institute of Advanced Industrial Science and Technology (AIST)
dataset submitter

jPOST dataset URI

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