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PXD062588-1
PXD062588 is an original dataset announced via ProteomeXchange.
Dataset Summary
| 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 |
Dataset History
| Revision | Datetime | Status | ChangeLog Entry |
|---|---|---|---|
| 0 | 2025-04-03 23:21:21 | ID requested | |
| ⏵ 1 | 2026-04-03 08:00:05 | announced |
Publication List
| Dataset with its publication pending |
Keyword List
| submitter keyword: AAV, glycosylation, glycoproteomics |
Contact List
| Atsushi Kuno | |
|---|---|
| lab head | |
| Hiroaki Sakaue | |
| contact affiliation | National Institute of Advanced Industrial Science and Technology (AIST) |
| dataset submitter | |
Full Dataset Link List
| jPOST dataset URI |
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