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<p>Sedimentation velocity analytical ultracentrifugation (SV-AUC) has become a workhorse technique for characterization and quantitation of AAV-based therapeutics, providing information on distribution of various AAV capsid species. However, SV-AUC results can be compromised by cell misalignment. In this study, we used an optical alignment system to investigate the combined effects of cell misalignment and rotor speed on the sedimentation behavior and measured distributions of AAV species. Our findings indicate that at high speeds even minor deviations in cell alignment can introduce substantial artifacts, leading to inaccurate quantitation of relative levels of different AAV capsid species by SV-AUC and potentially to misinterpretation of product quality.</p>

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Sedimentation velocity analytical ultracentrifugation (SV-AUC) for adeno-associated virus characterization: impact of cell alignment

  • 期刊论文

  • 发表日期：

    2026年3月14日

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  • Qianyi Luo,
  • Brandon L. Doyle,
  • Ivan L. Budyak

摘要

Sedimentation velocity analytical ultracentrifugation (SV-AUC) has become a workhorse technique for characterization and quantitation of AAV-based therapeutics, providing information on distribution of various AAV capsid species. However, SV-AUC results can be compromised by cell misalignment. In this study, we used an optical alignment system to investigate the combined effects of cell misalignment and rotor speed on the sedimentation behavior and measured distributions of AAV species. Our findings indicate that at high speeds even minor deviations in cell alignment can introduce substantial artifacts, leading to inaccurate quantitation of relative levels of different AAV capsid species by SV-AUC and potentially to misinterpretation of product quality.