| アイテムタイプ |
itemtype_ver1(1) |
| 公開日 |
2025-12-23 |
| タイトル |
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タイトル |
Analytical dissection of minor glycoforms and glycoprotein associations in rAAV preparations by multimodal glycoproteomics |
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言語 |
en |
| 著者 |
Kuno, Atsushi
Sakaue, Hiroaki
Koizumi, Sachiko
Tomioka, Azusa
Mizukado, Saho
Yamaguchi, Yuki
Fukuhara, Mitsuko
Tsunaka, Yasuo
Kaji, Hiroyuki
Uchiyama, Susumu
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| アクセス権 |
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アクセス権 |
embargoed access |
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アクセス権URI |
http://purl.org/coar/access_right/c_f1cf |
| 権利 |
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|
権利情報 |
This version of the article has been accepted for publication, after peer review (when applicable) and is subject to Springer Nature’s AM terms of use, but is not the Version of Record and does not reflect post-acceptance improvements, or any corrections. The Version of Record is available online at: http://dx.doi.org/10.1007/s00216-025-06042-4 |
|
言語 |
en |
| 内容記述 |
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内容記述タイプ |
Abstract |
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内容記述 |
Accurate glycan analysis of viral vectors is essential for evaluating pharmaceutical quality. Recent advances in mass spectrometry–based analytical technologies have achieved glycosylation detection in adeno-associated viruses (AAVs). However, because only a minor subpopulation (< 1%) of recombinant AAV (rAAV) particles may carry glycans or associate with glycoproteins, distinguishing genuine AAV glycosylation from that of co-purified glycoproteins remains technically challenging, highlighting the need for analytical strategies that minimize glycan misassignment and reliably identify glycoprotein interactions. Here, we present a multimodal glycoproteomic approach to discriminate rare glycosylation events on rAAV capsids from glycosylated host-derived proteins associated with the particles. We employed an ultrasensitive lectin microarray coupled with a broadly reactive anti-AAV antibody to detect O-glycan-binding lectin signals in several rAAV preparations. Notably, a distinct signal was observed for Urtica dioica agglutinin (UDA). Subsequent liquid chromatography-tandem mass spectrometry, combined with UDA-based dual enrichment at both protein and peptide levels, identified a divalently high-mannose N-glycosylated peptide derived from the host AAV receptor (AAVR). Monovalent high-mannose N-glycopeptides of AAVR and Mac-2 binding protein were additionally detected using single-step protein-level enrichment, indicating an avidity-driven UDA binding mechanism. However, no N-glycosylation was detected on the rAAV capsids themselves. These findings underscore the value of integrated multimodal glycoproteomic workflows for resolving low-abundance glycosylated species and offer new insights into host-derived hitchhiker glycoproteins that may affect rAAV characterization and quality control. |
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言語 |
en |
| 出版者 |
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出版者 |
Springer |
|
言語 |
en |
| 言語 |
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|
言語 |
eng |
| 資源タイプ |
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資源タイプresource |
http://purl.org/coar/resource_type/c_6501 |
|
タイプ |
journal article |
| 出版タイプ |
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|
出版タイプ |
AM |
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出版タイプResource |
http://purl.org/coar/version/c_ab4af688f83e57aa |
| 関連情報 |
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|
関連タイプ |
isVersionOf |
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|
識別子タイプ |
DOI |
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関連識別子 |
https://doi.org/10.1007/s00216-025-06042-4 |
| 収録物識別子 |
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収録物識別子タイプ |
PISSN |
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収録物識別子 |
1618-2642 |
| 書誌情報 |
en : Analytical and bioanalytical chemistry
巻 417,
号 23,
p. 5155-5170,
発行日 2025-09
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| ファイル公開日 |
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日付 |
2026-09-01 |
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日付タイプ |
Available |
Kuno_et_al_AnalBioanalChem_2025_revisedMS.pdf (1.9 MB)
