How to identify and quantify adeno-associated virus fill states using analytical ultracentrifugation

Recombinant adeno-associated viruses (AAV) are used extensively as a vector for gene therapy. AAV products are composed of a proteinaceous capsid that encapsulates the single-stranded DNA genome. Preparations of purified AAVs typically contain capsid species. These include empty capsids, partially packaged capsids with less than the intended DNA, fully packaged capsids representing the drug product, and occasionally, over-packaged capsids with more DNA than intended. Characterization and quantification of these species is necessary to ensure safety and efficacy of gene therapy treatments.

Recently, Maruno et al. published a comprehensive analysis using sedimentation velocity analytical ultracentrifugation (SV-AUC) to characterize AAV size distributions using multiple detection wavelengths. This allowed for physical separation of the species through ultracentrifugation and identification and quantification of well resolved species.

Dr. N. Karl Maluf, Director of the Biophysical Characterization Core at KBI Biopharma, will explores how to implement this analysis to identify various fill states in an AAV preparation.

Key learning objectives

• Learn the principles behind centrifugation and its advantages in isolating extracellular vesicles (EVs) from complex biological matrices
• Explore different sedimentation and density gradient centrifugation protocols, highlighting the critical parameters that impact EV yield and purity
• Understand the capabilities of analytical ultracentrifugation (AUC) to characterize EV purity, shape, and composition

Who should attend?

Industrial and academia researchers focused on AAVs

Certificate of attendance
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