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Abstract:
New designer cells as alternatives to well-established cell lines such as Vero or BHK are under consideration for vaccine production. A very successful example for such a designer cell line is Crucells’ human PER.C6 currently either used or evaluated for production of vaccines against Ebola, Malaria, West Nile disease and Influenza virus. Recently, ProBioGen AG has created a stable avian cell line (AGE1.CR) as substrate for vaccine production using adenovirus type 5 E1 genes on cells from duck retina. The CR line was further modified via stable expression of adenoviral pIX gene that encodes a structural protein involved in capsid stabilization. The CR and CR.pIX avian cell lines proliferate in suspension in serum free (SFM) or protein free media (PFM) and can be productively infected with different influenza strains. Another important application for CR and CR.pIX is production of modified vaccinia Ankara (MVA). Due to its high attenuation MVA does not proliferate in mammalian cells (with exception of BHK) and is produced in chicken embryo fibroblast with all disadvantages inherent in processes with primary cells.
Here, we present data for the cultivation of CR and CR.pIX cells in roller bottles and small scale bioreactors (STR, Wave). Surprisingly, the introduction of the pIX gene appears to impact the metabolism of CR cells. Differences between the two cell lines in metabolism (changes in consumption or release of glucose, lactate, glutamine, ammonia, glutamate and other amino acids) together with cell density in different media will be discussed. Data on the impact of the pIX gene on MVA yield will be shown and discussed. For influenza, virus replication yields and virus dynamics will be compared to those obtained for adherent Vero and MDCK cells. Furthermore, data for the production of various influenza virus strains (Influenza A (H1N1, H3N2), Influenza B) will be compared and glycosylation fingerprints of the viral HA membrane proteins will be shown.
