Cell Culture Process Development For AAV Vector Production In Suspension Cells

Recombinant adeno-associated virus (rAAV) vectors are common gene transfer vehicles in gene therapy research and in developing potential new therapies. To support the growing need for high-quality vector, scalable production processes are needed.
In the first set of experiments, we describe how to adapt adherent HEK293T/HEK293 cells to suspension culture in a new cell culture medium, evaluating cell growth by population doubling time, morphology, and viability. And we describe how to use a Design of Experiments (DoE) methodology to evaluate several parameters that could affect productivity.
In the second set of experiments, we describe how to optimize the transient transfection, which forms the basis for a scalable AAV production process in single-use bioreactors. We measured viral particles/L, viral genomes/L, and percentage full capsids.
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