Scalability comparison between 50 and 500 liter stirred tank bioreactor for production of rAAV viral vector

Abstract

Viral vectors are a new class of biologics which facilitate gene transfer and modification in living cells, potentially treating a multitude of conditions with genetic causes. Scalable manufacturing technologies are critical to ensuring these cutting-edge medicines can be produced in sufficient quantities to meet the needs of process development, clinical trials, and ultimately commercial manufacturing Mol. Ther. 2016; 3: 15049." data-url="http://dx.doi.org/10.1038/mtm.2015.49">[1]Ayuso E. Manufacturing of recombinant adeno-associated viral vectors: new technologies are welcome. Mol. Ther. 2016; 3: 15049.Ayuso E. Manufacturing of recombinant adeno-associated viral vectors: new technologies are welcome. Mol. Ther. 2016; 3: 15049.Ayuso E. Manufacturing of recombinant adeno-associated viral vectors: new technologies are welcome. Mol. Ther. 2016; 3: 15049.. As viral vector-based products have only relatively recently received regulatory approval, public information on scalable optimization of these processes is very limited. Abeona Therapeutics is a gene therapy company developing novel gene replacement therapies for rare inherited diseases. These conditions can impact development and limit both quality of life and/or life expectancy et al. Estimation of impact of RPE65-mediated inherited retinal disease on quality of life and the potential benefits of gene therapy. Br. J. Ophthalmol. 2019; 103(11): 1610–14. " data-url="http://dx.doi.org/10.1136/bjophthalmol-2018-313089">[2]Lloyd A, Piglowska N, Ciulla T et al. Estimation of impact of RPE65-mediated inherited retinal disease on quality of life and the potential benefits of gene therapy. Br. J. Ophthalmol. 2019; 103(11): 1610–14. . These transformative medicines can be used to replace a defective gene with a functional copy, silence a defective gene or even directly edit genes Nat. Rev. Drug Discov. 2019; 18(5): 358–78. " data-url="http://dx.doi.org/10.1038/s41573-019-0012-9">[3]Wang D, Tai PWL, Gao G. Adeno-associated virus vector as a platform for gene therapy delivery. Nat. Rev. Drug Discov. 2019; 18(5): 358–78. , J. Virol. 1999; 73(5): 3994–4003. " data-url="http://dx.doi.org/10.1128/jvi.73.5.3994-4003.1999">[4]Xiao W, Chirmule N, Berta SC, McCullough B, Gao G, Wilson JM. Gene therapy vectors based on adeno-associated virus type 1. J. Virol. 1999; 73(5): 3994–4003. . We evaluated the Pall Allegro™ STR bioreactor family as an rAAV vector production platform and evaluated the scalability of the PEI-mediated transfection manufacturing process for rAAV at the 50 L and 500 L working volume. Process scalability was evaluated based on cell growth, metabolic profile, and vector production. This testing demonstrates that control of key process parameters enables a scalable vector production process between the 50 L and 500 L scale using Allegro STR single use bioreactors.