Impact of Low pH Affinity Chromatography Elution on AAV Transduction
Presented by: René Gantier, Ph.D. (Repligen)
Presented Live: October 22, 2025
Affinity chromatography has become a highly effective and scalable method for purifying adeno-associated virus (AAV) particles in gene therapy manufacturing. However, the use of acidic elution conditions (pH 2–3) continues to raise concerns about potential capsid instability and loss of infectivity, even when followed by immediate pH neutralization.
This webinar investigates the impact of low pH elution on AAV capsid integrity and biological activity. Using a combination of size exclusion chromatography coupled with multi-angle light scattering (SEC-MALS) and functional assays, we assess capsid aggregation and structural stability across three AAV serotypes—AAV5, AAV8, and AAV9—purified under various elution conditions and incubated at different temperatures. Functional infectivity is further evaluated using cell-based transduction assays.
We also explore the downstream effects of additional purification steps, including tangential flow filtration (TFF), and their role in promoting virus aggregation. Our findings offer valuable insights into the robustness of AAV particles under acidic conditions and underscore the importance of advanced analytical tools like SEC-MALS.