Continuous Production of Recombinant Adeno-associated Viral Vectors via Transient Transfection of HEK293 Cells in Perfusion Bioreactor

• Published in: Computer Aided Chemical Engineering Vol. 53; pp. 2587 - 2592
• Main Authors: Park, Damdae, Nguyen, Tam N.T., Sangerman, José, Srinivasan, Prasanna, Ou, Rui Wen, Katsikis, Georgios, Hong, Moo Sun, Barone, Paul W., Neufeld, Caleb, Wolfrum, Jacqueline M., Springs, Stacy L., Sinskey, Anthony J., Braatz, Richard D.
• Format: Book Chapter
• Language: English
• Published: 2024
• Subjects: continuous production; gene therapy; transient transfection; transient transfection; gene therapy; continuous production
• ISBN: 9780443288241; 0443288240
• ISSN: 1570-7946
• DOI: 10.1016/B978-0-443-28824-1.50432-4

Gene therapy offers a promising approach to remedy genetic deficiencies by introducing missing genes into the patient's body. Recombinant adeno-associated virus (rAAV) is one of the most widely used gene carriers, whose therapeutic doses reach up to ~1015 viral genomes (vg) per patient. However, existing technology imposes limitations on rAAV production, necessitating a large-scale and cost-effective manufacturing process. In this paper, we propose a continuous manufacturing process and a model-based control strategy for transient transfection of HEK293 cells. A dynamic model of the system is developed and utilized for soft sensing of the component concentrations in cell culture. Based on which, key operational decisions including transfection timing, plasmid dosages, and perfusion rates are made. We demonstrate that the proposed system and control strategy efficiently produce rAAV at a scale comparable to the conventional batch process of the same volume. Recommendations and insights are provided for further process intensifications and optimizations of the proposed system.