Optimization Of AAV Vector Production Using Statistical Design Of Experiment

By XingchengChen, Nida S. Zubairy, Timothy Ryan, Dogan Ornek, Tao Xiang, Chi-Ming Yu, W. Roy Lin, Chih-Cheng Chen, Boston Institute of Biotechnology, Southborough, MA 01772

Plasmid-mediated transient transfection in HEK293 cells is the quickest AAV expression system to develop and easy to scale, but titers typically sit at 10^9 to 10^10 vg/mL, which limits its fit for large-scale production. This poster from researchers at Boston Institute of Biotechnology takes a statistical approach to that gap. Starting from a risk assessment, the team screened six process parameters in a 17-run Definitive Screening Design built in JMP, then ran a confirmation experiment. Three critical process parameters emerged, along with operating ranges intended for scale-up, and the confirmation titer exceeded the model's prediction. The poster also covers the design matrix, the analytical workflow, and the parameter-titer relationships. Review the design and results to see which variables carried the most weight.