Language
English
Publication Date
9-1-2023
Journal
Nanomedicine
DOI
10.2217/nnm-2023-0156
PMID
37877696
PMCID
PMC10652292
PubMedCentral® Posted Date
10-25-2023
PubMedCentral® Full Text Version
Post-Print
Abstract
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.
Keywords
Capsid, Dependovirus, Ultracentrifugation, Genetic Vectors, Microscopy, Electron, Transmission, adeno-associated virus, analytical buoyant density gradient equilibrium, gene therapy, multi-wavelength analytical ultracentrifugation, sedimentation velocity, UltraScan, viral capsid quantification, viral vector
Published Open-Access
yes
Recommended Citation
Henrickson, Amy; Ding, Xiaozhe; Seal, Austin G; et al., "Characterization and Quantification of Adeno-Associated Virus Capsid-Loading States by Multi-Wavelength Analytical Ultracentrifugation With UltraScan" (2023). Faculty and Staff Publications. 1171.
https://digitalcommons.library.tmc.edu/baylor_docs/1171
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