Comparison of manufacturing techniques for adenovirus production

We have compared three different production methods, which may be suitable for the large scale production of adenovirus vectors for human clinical tri als. The procedures compared 293 cells adapted to suspension growth in seru m-free medium in a stirred tank bioreactor, 293 cells on microcarriers in s erum-containing medium in a stirred tank bioreactor, and 293 cells grown in standard tissue culture plasticware. With a given virus, yields varied bet ween 2000 and 10,000 infectious units/ cell. The stirred tank bioreactor ro utinely produced between 4000 and 7000 infectious units/ cell when 293 cell s were grown on microcarriers. The 293 cells adapted to suspension growth i n serum-free medium in the same stirred tank bioreactor yielded between 200 0 and 7000 infectious units/ cell. Yields obtained from standard tissue cul ture plasticware were up to 10, 000 infectious units/ cell. Cell culture co nditions were monitored for glucose consumption, lactate production, and am monia accumulation. Glucose consumption and lactate accumulation correlated well with the cell growth parameters. Ammonia production does not appear t o be significant. Based on virus yields, ease of operation and linear scala bility, large-scale adenovirus production seems feasible using 293 cells (a dapted to suspension/serum free medium or on microcarriers in serum contain ing medium) in a stirred tank bioreactor.