Assessing Critical Quality Attributes Of AAVs Including Aggregation And Capsid Load Using A Biozen dSEC-7 Column

By Claire Butré, PhD, Jennifer Bouchenna, PhD, Damien Mouvet, Arnaud Delobel, PhD, and Bryan Tackett, PhD

This content is brought to you by Phenomenex, a Danaher Operating Company.

Adeno-associated viral vectors (AAVs) are critical components for delivering therapeutic genes, but their quality control requires rigorous assessment of several critical quality attributes (CQAs). Aggregation and the presence of empty capsids are two such impurities that must be controlled, as they can negatively impact the therapeutic product's efficacy and safety.

Traditionally, determining the crucial full-to-empty capsid ratio relies on analytical ultracentrifugation, a reproducible method but one that is sample-intensive and challenging to implement in a GMP setting. This application note highlights an alternative, Size Exclusion Chromatography coupled with Multi-Angle Light Scattering (SEC-MALS), which offers enhanced throughput by eliminating sample preparation. SEC-MALS enables reliable resolution of AAV monomers and aggregates and allows for accurate determination of capsid and DNA molar masses, particle concentration, and the percentage of full AAVs in a single analysis. Discover how this approach successfully characterized AAV5 and AAV8 serotypes.