Recombinant adeno-associated viral vectors (rAAV) are increasingly used as the vector of choice in gene therapy. The purification process of rAAVs involves cell lysis, clarification of the lysed cells to separate the supernatant containing the rAAV, benzonase® treatment, followed by several purification steps. The final step is an ultrafiltration/diafiltration step to concentrate the viral vector and place it in the formulation buffer matrix.<br/>While the ultrafiltration behavior of proteins and monoclonal antibodies are very well understood, rAAV transport in ultrafiltration membrane modules are not. This work provides data to demonstrate the impact of membrane molecular weight cut-off, transmembrane pressure, crossflow-rate and the choice of constant TMP-mode or constant permeate flux-mode on the process performance and product quality (recovery of rAAV and aggregation). Several process runs using pressure-dependent ultrafiltration and pressure-independent ultrafiltration are compared to identify the most robust operating condition.<br/>Finally, the choice of reusable ultrafiltration modules is compared against single-use ultrafiltration modules keeping product quality, cost of goods and process performance as performance indicators for commercial rAAV manufacturing.<br/>

Ultrafiltration behavior of adeno-associated viral vectors: Understanding the implications of choice of process parameters and device format on commercial manufacturing

ACS Fall 2020 Virtual Meeting

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