Application Note: Centrifuge Free Clarification and Harvest of Adeno-associated Viruses (AAV)
Authors: Frederik Meierrieks and Andreas Pickl
This study aimed to evaluate the applicability of Sartoclear Dynamics® Lab Filter Aid, containing diatomaceous earth (DE), for clarifying crude lysates of adeno-associated virus (AAV) produced by HEK293 suspension cells. Filtration using Sartoclear Dynamics® Lab Filter Aid proved to be a viable clarification method for AAV2, AAV5 and AAV8. For standard AAV production processes, AAV particle recovery in the filtrate was >95%. Even difficult-to-handle serotypes such as AAV2 and lysates containing lower AAV particle titers could be clarified by Sartoclear Dynamics® Lab filtration with viral recoveries of >80%.
Overview
This study outlines a successful optimization strategy that utilized a Design of Experiments (DoE) approach to increase the yield of AAV2 to over 95%. Moreover, the use of Sartoclear Dynamics® Lab Filter Aid reduced manual handling time 3-fold and increased the filter capacity 3.5-fold compared to the standard laboratory method of filtration combined with a prior centrifugation step.
- Page Count: 8
- Read Time: 14 minutes
- Last Updated: May 5, 2023
Key Takeaways:
- Why Sartoclear Dynamics® Lab filtration is a viable approach to clarifying AAV crude cell lysates.
- The impact of Sartoclear Dynamics® Lab on turbidity removal, adeno-associated virus (AAV) yield, filter capacity and manual handling times.
- The effect of diatomaceous earth (DE) concentration and DE incubation time on AAV2 retention and turbidity removal in a Design of Experiments (DoE) study.
This Resource is Designed for:
- Lab Researchers
Applications Supported:
- Life Science Research
- Clarification
- Viral vectors
- Gene therapy
- Diatomaceous earth (DE)
- Adeno-associated virus (AAV)
