This application note describes a process for adenovirus production, from upstream cell culture to downstream purification, using modern tools and technologies. To demonstrate robustness of the upstream process, virus production in suspension adapted cells was performed in both rocking and stirred-tank bioreactors. The downstream process was carefully optimized to meet stringent regulatory demands on product purity and quality. Novel analytical methods were used in parallel with established techniques for comparison and to ensure accurate monitoring of the processed material. The novel process is easily scaled and compatible with both single-use and steamable hard-piped process equipment.