Impact of Low pH Affinity Chromatography Elution on AAV Transduction
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.
Key Learning Objectives:
- Impact of low pH affinity chromatography elution on AAV transduction
- Strategies to mitigate risks of capsid inactivation and destabilization after low pH affinity capture
- Applications of SEC-MALS for AAV characterization
Presenter: René Gantier, Ph.D. (Senior Director R&D, Advanced Bioprocess Applications, Repligen)
René Gantier is the Senior Director R&D, Advanced Bioprocess Applications at Repligen. He has more than 20 years of experience developing production and purification processes for biotherapies. In his current role at Repligen, he leads the development of advanced bioprocess applications and technologies with a focus on next generation cell and gene therapy manufacturing processes. Before joining Repligen, he was leading the Biotech Process R&D team at Pall Life Sciences developing end-to-end upstream-downstream continuous bioprocessing solutions for mAbs and cell/gene therapies. René holds a MSc in Cell Biology from the University of Nantes (France) and a Ph.D. in Biochemistry from the University of Rouen (France).
