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Process evaluation of apoptosis resistant cells expressing a recombinant antibody

The impact of bcl-2 on culture viability, apoptosis, product yield / productivity, product recovery and other parameters could be demonstrated under non-optimised culture conditions (e.g. standard medium, feed). This has been performed at incubator scale (T-flasks, used for inoculum), standard fed-batch cultivation (at a 2 L scale) as well as for enhanced fed-batch cultivation using dialysis (2 L scale).

As a model system the cell lines NS0 6A1 (100)3 (control cell line, not transfected with bcl-2) and NS0 6A1 bcl-2 (transfected with bcl 2) have been used. Consistent results have been obtained at all stages. The improvement of the bcl-2 over-expression can be clearly seen in the prolonged growth phase and a higher maximum cell density resulting in a higher antibody production. The final product has been analysed with respect to purity with particular attention to the degree of contamination with cellular DNA.

Concerning the different processes, improved growth and product formation characteristics were obtained using dialysis-fed-batch cultivation in comparison to standard fed-batch cultivation. In addition, the successful performance of the developed fed-batch control could be demonstrated.

The results have already been published:
Frahm, B., Lane, P., Märkl, H., Pörtner, R. (2003): Improvement of a mammalian cell culture process by adaptive, model-based dialysis fed-batch cultivation and suppression of apoptosis, Bioprocess and Biosystems Engineering 26, 1-10.

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