Hydrodynamic characterization of a vesicular stomatitis virus-based oncolytic virus using analytical ultracentrifugation

Determination of the size, density, and mass of viral particles can provide valuable information to support process and formulation studies in clinical development. Analytical ultracentrifugation (AUC), as a first principal method, has been shown to be a beneficial tool for the characterization of t...

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Bibliographic Details
Published in:European biophysics journal 2023-07, Vol.52 (4-5), p.379-386
Main Authors: Wawra, Simon, Kessler, Sophia, Egel, Arina, Solzin, Johannes, Burkert, Oliver, Hochdorfer, Daniel
Format: Article
Language:English
Subjects:
Biochemistry
Biological and Medical Physics
Biomedical and Life Sciences
Biophysics
Cell Biology
Density gradients
Information processing
Life Sciences
Membrane Biology
Molecular weight
Nanoparticles
Nanotechnology
Neurobiology
Oncolysis
Original
Original Article
Specific volume
Stomatitis
Ultracentrifugation
Viruses
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Summary:Determination of the size, density, and mass of viral particles can provide valuable information to support process and formulation studies in clinical development. Analytical ultracentrifugation (AUC), as a first principal method, has been shown to be a beneficial tool for the characterization of the non-enveloped adeno associated virus (AAV). Here, we demonstrate the suitability of AUC for the challenging characterization of a representative for enveloped viruses, which usually are expected to exhibit higher dispersity than non-enveloped viruses. Specifically, the vesicular stomatitis virus (VSV)-based oncolytic virus VSV-GP was used to evaluate potential occurrence of non-ideal sedimentation by testing different rotor speeds and loading concentrations. The partial specific volume was determined via density gradients and density contrast experiments. Additionally, nanoparticle tracking analysis (NTA) was used to determine the hydrodynamic diameter of VSV-GP particles to calculate their molecular weight via the Svedberg equation. Overall, this study demonstrates the applicability of AUC and NTA for the characterization of size, density, and molar mass of an enveloped virus, namely VSV-GP.
ISSN:0175-7571
1432-1017
DOI:10.1007/s00249-023-01649-w