Use Of High-Throughput And DoE Tools For Robust Development Of DSP Technologies For AAV

By F. Bollmann, M. Noverraz, P. Nestola, G. Pressac, A. Znidarsic, M. Leskovec, T. Kostelec, U. Dippel, Y. Volz, A. Thiefes, A. Martel, T. Faria, S. Mendes, A. Nascimento, M. Moleirinho, R. Silva, A. Roldão, and C. Peixoto

Adeno-associated viral vectors are a common vehicle for gene therapy delivery. However, producing them remains a challenge for manufacturers due to the complexity of developing platform production processes and the inefficiencies of downstream processing. As the potential for diversifying adeno-associated virus (AAV) capsid serotypes grows for gene therapy applications, there is a heightened interest in a downstream processing platform capable of purifying various serotypes with consistent recovery and purity. This could shorten process development timelines and simplify manufacturing complexities.

The focus of this study was to optimize the three primary downstream processing steps for AAV particle purification: harvest clarification, tangential flow filtration (TFF), and capture chromatography. To achieve this, a design of experiments (DoE) methodology and screening devices were employed to screen several materials and process parameters. Continue reading to see the results.