Comparison of Manufacturing Techniques for Adenovirus Production

Abstract

We have compared three different production methods which may be suitable for the large-scale production of adenovirus vectors for human clinical trials. The procedures compared 293 cells adapted to suspension growth in serum-free medium in a stirred tank bioreactor, 293 cells on microcarriers in serum containing medium in a stirred tank bioreactor, and standard tissue culture plasticware. With a given virus, yields varied between 2000 and 12,000 infectious units (iu)/cell. The stirred tank bioreactor routinely produced between 4000 and 7000 iu/cell when 293 cells were grown on microcarriers. 293 cells adapted to suspension growth in serum free medium in the same stirred tank bioreactor yielded between 2000 and 7000 iu/cell. Yields obtained from standard tissue culture plasticware were up to 12,000 iu/cell. Cell culture conditions were monitored for glucose consumption, oxygen utilization, lactate production, and ammonia accumulation. Oxygen utilization rate, glucose consumption and lactate accumulation correlated well with the cell growth parameters. Ammonia production does not appear to be significant. Based on virus yields, ease of operation and linear scalability, large scale adenovirus production seems feasible using 293 cells (adapted to suspension/serum free medium or on microcarriers in serum containing medium) in a stirred tank bioreactor.