706. Consistent Viral Vector Manufacturing for Phase III Using iCELLis 500 Fixed-Bed Technology

Abstract

Single use disposable technology faces further challenges in producing viral vectors in sufficient large quantities especially where adherent cells are needed. Scaling up the adherent system has proven to be troublesome. The PALL iCELLis® disposable fixed-bed bioreactors offer a novel option for viral vector manufacturing in large quantities in an adherent environment. We have made process development in iCELLis Nano, where the cultivation area varies between 0.53 - 4 m2, after which we went forward to iCELLis 500, where the cultivation volume can be upgraded to 500 m2 (66 - 500 m2). iCELLis 500 has proven to be ideal to satisfy upstream demand and large-scale downstream purification process was developed to supply high quality recombinant adenovirus based gene products in our fully-licensed GMP manufacturing facility for pre-clinical and clinical trials The process is initiated by HEK293 cell mass expansion in suspension mode using Biostat® CultiBag RM (Sartorius Stedim Biotech S.A.) bioreactor. The expanded suspension cell mass is inoculated into iCELLis 500 for further expansion in adherent mode. This is infected by Working Viral Seed Stock and subsequent virus is released from the infected cells by chemical lysis. Downstream process contains Benzonase digestion, clarification, concentration and conditioning by crossflow ultrafiltration, capture and polishing by anion exchange chromatography, and final concentration and formulation is achieved using crossflow filtration. All product contact parts are fully disposable. Several batches have been produced with consistent results. Further validation of the process for commercial manufacturing is currently ongoing. We established a scalable, large-scale manufacturing process to supply high quality recombinant adenovirus based gene products in our fully-licensed GMP manufacturing facility for clinical trials. Alongside this we have validated assays which are providing a relevant quantitative measure of biologic function of the vector and demonstrating the quality and comparability of drug product batches in Phase 3 and commercial use.