Successful Development of a Robust Adenovirus Production Process Primarily Relies on a Better Understanding of Packaging Cell Line Physiology and Vector Replication Kinetics

Abstract

This report will focus on demonstrating how the fundamental understanding of complementing cell line physiology and metabolism, viral infection kinetics, culture conditions and viral particle stability is a prerequisite to defining optimal operating conditions for improving final adenoviral yield and activity. Steps involved in developing and optimising a robust integrated process for adenovirus production using HEK-293 cells cultured in suspension and serum free medium will be reviewed using data generated in batch, fed-batch and perfusion culture modes from 3, 20L and 60L bioreactor runs. Development of new monitoring tools (in-situ GFP probe, permittivity) and quantification techniques (HPLC-determination of total particles) greatly contributed to acceleration of process development. Online monitoring of physiological parameters such as respiration and biovolume during the course of infection/production allowed a real-time supervision and control of critical phases of the process.