Quantitation of AAV in a dual-vector system using SV-AUC.
| Autor: | Yarawsky AE; BioAnalysis, LLC, 3401 I Street Suite 206, Philadelphia, PA 19134, USA., Ciatto C; Decibel Therapeutics, Inc., 1325 Boylston Street Suite 500, Boston, MA 02215, USA., Slade P; Decibel Therapeutics, Inc., 1325 Boylston Street Suite 500, Boston, MA 02215, USA., Figueroa NI; BioAnalysis, LLC, 3401 I Street Suite 206, Philadelphia, PA 19134, USA., Burgner JW 2nd; BioAnalysis, LLC, 3401 I Street Suite 206, Philadelphia, PA 19134, USA., DeLion MT; BioAnalysis, LLC, 3401 I Street Suite 206, Philadelphia, PA 19134, USA., Paul LN; BioAnalysis, LLC, 3401 I Street Suite 206, Philadelphia, PA 19134, USA. Electronic address: lpaul@bioanalysisllc.com. |
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| Jazyk: | angličtina |
| Zdroj: | Journal of pharmaceutical sciences [J Pharm Sci] 2024 Oct 28. Date of Electronic Publication: 2024 Oct 28. |
| DOI: | 10.1016/j.xphs.2024.10.049 |
| Abstrakt: | Sedimentation velocity analytical ultracentrifugation (SV-AUC) has become the "gold standard" for characterization of the empty, partial, and full capsids of gene therapy products (e.g., AAV and Adenovirus vectors). Other techniques, such as SEC-MALS, TEM, and mass photometry, are commonly used for capsid quantitation, however, the resolving power of these techniques is lacking. In this body of work, SV-AUC was implemented in the characterization of a dual-vector AAV system where the difference in packaged genomes was ∼400 nucleotides. The instrument parameters and SV-AUC analysis were optimized to accurately quantitate both AAV vectors with less than 8% error and with highly correlated linearity (R 2 > 0.99) as compared to ddPCR. The results of this work highlight the resolution and accuracy of dual-vector capsid quantitation by SV-AUC and demonstrate the use of the powerful Bayesian analysis implemented in the SEDFIT analysis software. Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: All authors are employees or consultants of BioAnalysis, LLC or Decibel Therapeutics, Inc. (now part of Regeneron). This work was funded by BioAnalysis, LLC and Decibel Therapeutics, Inc. (Copyright © 2024 American Pharmacists Association. Published by Elsevier Inc. All rights reserved.) |
| Databáze: | MEDLINE |
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