Characterization and quantification of adeno-associated virus capsid-loading states by multi-wavelength analytical ultracentrifugation with UltraScan
Aim: We present multi-wavelength (MW) analytical ultracentrifugation (AUC) methods offering superior accuracy for adeno-associated virus characterization and quantification. Methods: Experimental design guidelines are presented for MW sedimentation velocity and analytical buoyant density equilibrium AUC. Results: Our results were compared with dual-wavelength AUC, transmission electron microscopy and mass photometry. In contrast to dual-wavelength AUC, MW-AUC correctly quantifies adeno-associated virus capsid ratios and identifies contaminants. In contrast to transmission electron microscopy, partially filled capsids can also be detected and quantified. In contrast to mass photometry, first-principle results are obtained. Conclusion: Our study demonstrates the improved information provided by MW-AUC, highlighting the utility of several recently integrated UltraScan programs, and reinforces AUC as the gold-standard analysis for viral vectors.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:18 |
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| Enthalten in: |
Nanomedicine (London, England) - 18(2023), 22 vom: 14. Sept., Seite 1519-1534 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Henrickson, Amy [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 15.11.2023 Date Revised 22.04.2024 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.2217/nnm-2023-0156 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM363699279 |
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| 100 | 1 | |a Henrickson, Amy |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Characterization and quantification of adeno-associated virus capsid-loading states by multi-wavelength analytical ultracentrifugation with UltraScan |
| 264 | 1 | |c 2023 | |
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| 500 | |a Date Completed 15.11.2023 | ||
| 500 | |a Date Revised 22.04.2024 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Aim: We present multi-wavelength (MW) analytical ultracentrifugation (AUC) methods offering superior accuracy for adeno-associated virus characterization and quantification. Methods: Experimental design guidelines are presented for MW sedimentation velocity and analytical buoyant density equilibrium AUC. Results: Our results were compared with dual-wavelength AUC, transmission electron microscopy and mass photometry. In contrast to dual-wavelength AUC, MW-AUC correctly quantifies adeno-associated virus capsid ratios and identifies contaminants. In contrast to transmission electron microscopy, partially filled capsids can also be detected and quantified. In contrast to mass photometry, first-principle results are obtained. Conclusion: Our study demonstrates the improved information provided by MW-AUC, highlighting the utility of several recently integrated UltraScan programs, and reinforces AUC as the gold-standard analysis for viral vectors | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, U.S. Gov't, Non-P.H.S. | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a UltraScan | |
| 650 | 4 | |a adeno-associated virus | |
| 650 | 4 | |a analytical buoyant density gradient equilibrium | |
| 650 | 4 | |a gene therapy | |
| 650 | 4 | |a multi-wavelength analytical ultracentrifugation | |
| 650 | 4 | |a sedimentation velocity | |
| 650 | 4 | |a viral capsid quantification | |
| 650 | 4 | |a viral vector | |
| 700 | 1 | |a Ding, Xiaozhe |e verfasserin |4 aut | |
| 700 | 1 | |a Seal, Austin G |e verfasserin |4 aut | |
| 700 | 1 | |a Qu, Zhe |e verfasserin |4 aut | |
| 700 | 1 | |a Tomlinson, Lauren |e verfasserin |4 aut | |
| 700 | 1 | |a Forsey, John |e verfasserin |4 aut | |
| 700 | 1 | |a Gradinaru, Viviana |e verfasserin |4 aut | |
| 700 | 1 | |a Oka, Kazuhiro |e verfasserin |4 aut | |
| 700 | 1 | |a Demeler, Borries |e verfasserin |4 aut | |
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