672. iCELLisTM Fixed-Bed Technology for Adherent Cells Is an Efficient Scalable System for Viral Vector Production Applications

Abstract

To date many early phase gene therapy trials have been successful. However, phase III and commercial phase have brought further challenges in producing viral vectors in sufficient large quantities. Upscaling the process on suspension cells is feasible, but many viral production applications are still applicable only in adherent settings. Scaling up the adherent system has proven to be troublesome and costly. The PALL iCELLisTM disposable fixed-bed bioreactors offer an efficient option for viral vector manufacturing in large quantities in an adherent environment. In iCELLisTM Nano, the cultivation area 0.53-4 m2 for smaller batches, is ideal for process development purposes. In iCELLIs500 the cultivation area varies between 66 and 500 m2 and is ideal to satisfy demand for phase3/commercial requirements.We have optimized adenovirus type 5 manufacturing using iCELLisTM Nano. HEK293 cell cultivation, infection and harvest of the virus by lysing the cells inside the bioreactor were efficient, reaching total yield of 3.4 × 10^14 vp/batch. When upscaling the process into 100 m2 cultivation area with iCELLis500, 1.2 × 10^16 vp/batch were produced.iCELLisTM technology is applicable also to other vector types which require for example plasmid transfection. Virus can also be harvested by perfusion from the medium. Lentiviral vector production in 293T cells was tested in iCELLisTM Nano and we achieved high plasmid transfection efficiency, leading to the comparable titers and productivity as in flasks. To conclude, iCELLisTM equipment has provided us an efficient way to manufacture large batches of different kinds of gene therapy products suitable for large preclinical animal models and up to phase III trial and beyond.