How To Identify And Quantify Adeno-Associated Virus Fill States Using Analytical Ultracentrifugation

Recombinant adeno-associated viruses (AAVs) are workhorses in gene therapy, acting as vectors to deliver genetic material. However, AAV preparations are a mixed bag. These viral packages, composed of a protein shell (capsid) encapsulating single-stranded DNA, come in various fill states. Empty capsids, partially filled capsids (missing some DNA), the ideal fully-packaged capsids (the drug itself), and even over-stuffed capsids (containing more DNA than intended) can all be present. To ensure gene therapy treatments are safe and effective, it's critical to identify and quantify these different AAV species.

Maruno et al. recently published a powerful method using sedimentation velocity analytical ultracentrifugation (SV-AUC) with multiwavelength detection to analyze AAV size distributions. This technique physically separates the AAV species through ultracentrifugation, allowing researchers to not only distinguish them but also determine their quantities. This webinar delves into how to implement this analysis to pinpoint the various fill states present in AAV preparations.

Key Takeaways:

• Multiwavelength AUC analysis of AAV preparations offers a robust approach to identifying and quantifying different AAV fill states.
• Explore the limitations of other techniques like SEC-MALS for characterizing AAV samples.
• Gain insights into the usefulness of complementary methods like analytical CsCl density gradient analysis for AAV characterization.