Three Ways to Quantify Adeno-Associated Viruses with Analytical Ultracentrifugation
Adeno-associated viruses (AAVs) are leading vectors for recombinant DNA delivery in gene therapy. However, ensuring the safety and efficacy of these therapies requires rigorous characterization of viral loading states—specifically distinguishing between empty, partial, full, and overfilled capsids.
Analytical ultracentrifugation (AUC) stands out as a premier, first-principles, label-free biophysical method capable of directly measuring these critical attributes. By operating without calibration standards or assumptions about particle shape, AUC delivers highly accurate, quantitative data in matrix-replicated environments.
Key Takeaways
This whitepaper explores the three primary modes of AUC optimized for AAV characterization, allowing researchers to choose the best fit for their sample volumes, resolution needs, and workflows:
- Sedimentation Velocity (SV-AUC): The “gold standard” for quantifying empty/full capsid ratios, identifying aggregates, and evaluating overall sample purity directly in formulation buffers.
- High-Speed Sedimentation Velocity (hs-SV-AUC): An optimized, rapid application that slashes runtime to 20–45 minutes while offering the ultra-high resolution needed to separate dual-vector AAV mixtures varying by as few as 300 nucleotides.
- Density Gradient Equilibrium (DGE-AUC): An orthogonal equilibrium technique utilizing cesium chloride ($\text{CsCl}$) gradients that reduces sample requirements by approximately 20-fold, making it ideal for scarce or low-concentration samples.
Who Should Read This?
This document is an indispensable resource for biopharma researchers, process development scientists, and quality control specialists looking to optimize their gene therapy analytics, ensure product potency, and accelerate regulatory decision-making.
White Paper from 