182 - Industrialization of AdenoVirus production and purification with the iCELLis® 500 single-use bioreactor

Abstract

Diabetes is a major global health problem with the WHO reporting over 422 million affected persons and 3.7 million diabetes related deaths annually worldwide. Orgenesis is developing a cell therapy approach that genetically modifies a patient's own liver cells to secrete insulin. To bring this approach into pharmaceutical development, Orgenesis and Pall have combined their respective expertise to develop a manufacturing strategy for large scale adenoviral vector production in the packed-bed iCellis® 500 single-use bioreactor together with the development of a downstream purification industrialized process. In this study, we describe scale-up of the optimized adenovirus serotype 5 production process that was developed using a predictive small scale iCellis® Nano bioreactor. By optimizing culture and infection parameters such as HEK293 cell seeding density, multiplicity of infection, time of infection, day of harvest and media circulation parameters, yield was increased from 1.6E9 to 1.4E10 infectious virus particles per cm2 fixed bed. This yield makes the iCellis® bioreactor a promising scalable technology for the production of adenovirus products. The current density gradient based purification method for adenovirus is a time consuming, inefficient and non-manufacturing process. Here, an optimized rapid AEX membrane chromatography step was used for purification. Following depth and sterile filtration, clarified harvest is processed over a Mustang Q membrane in bind/elute mode. Eluted material is immediately concentrated and buffer exchanged into the final formulation buffer. Final product is then sterile filtered and vialed for potency studies. Purified adenovirus hPDX-1 was fully functional and successfully transduced the target liver derived cells. The next step of the study will incorporate the viral trans-differentiation step into the cGMP autologous patient cell expansion process in Pall's Xpansion® 200 bioreactor.