Study of a perfusion process of Chinese Hamster Ovary cells by ATF filtration in bioreactor
2009 (English)Other (Other academic)
Perfusion is a mode of operation where a continuous replacement of the conditioned medium by fresh medium is operated. It has the advantage of allowing high cell densities. This mode of operations is also required for some instable proteins since the cell-free supernatant containing the product of interest is immediately stored at low temperature where the proteolysis is not active. The ATF filtration device has been designed to perfuse mammalian cell cultivation process. The cell broth circulation back and forward in the filter prevents the filter clogging and the design ensures a low shear not damageable for the cells.
The purpose of the present work was to develop and study a perfusion process of Chinese Hamster Ovary cells producing a monoclonal antibody by ATF filtration in a 2 L working volume bioreactor. A serum-free medium was used (Irvine Scientific IS CHO-CD). Cell densities above 40 x 106 cells/mL were obtained. These high cell densities were challenging for the aeration. Pure oxygen aeration by large bubbles from an open tube resulted in satisfying oxygenation until 25 to 30 x 106 cells/mL but became limiting at higher cell densities due to the low kLa of these bubbles and the small liquid height. At higher cell densities, a porous sparger with pure oxygen was used either alone or in combination with the open tube aeration. The performances of the perfusion by ATF filtration, the aeration and the use of anti-foam are presented and discussed.
Place, publisher, year, edition, pages
ATF, perfusion, CHO Cells
Research subject SRA - Molecular Bioscience
IdentifiersURN: urn:nbn:se:kth:diva-87462OAI: oai:DiVA.org:kth-87462DiVA: diva2:501675
Abstract included in the Proceedings of the 21st ESACT conference (European Society for Animal Cell Technology), , Dublin, Ireland, June 7-10, 2009. QC 201202232012-02-142012-02-142012-03-21Bibliographically approved