Establishment of a mink enteritis vaccine production process in stirred-tank reactor and Wave® Bioreactor microcarrier culture in 1–10L scale

• Published in: Vaccine 2007-05, Vol.25 (20), p.3987-3995
• Main Authors: Hundt, B., Best, C., Schlawin, N., Kaßner, H., Genzel, Y., Reichl, U.
• Format: Article
• Language: English
• Subjects: Ammonium; Bioreactors; Cell growth; Cultivation; Glucose; Immunization; Infections; Manufacturing; Mink enteritis virus; Propagation; Vaccines; Viral infections
• ISSN: 0264-410X; 1873-2518
• DOI: 10.1016/j.vaccine.2007.02.061

A scale-up and process optimization scheme for the growth of adherent embryonic feline lung fibroblasts (E-FL) on microcarriers and the propagation of a mink enteritis virus (MEV) strain for the production of an inactivated vaccine is shown. Stirred-tank cultivations are compared with results obtained from Wave®Bioreactors. Transfer from a roller bottle-based production process into large-scale microcarrier culture with starting concentrations of 2g/L Cytodex(TM) 1 microcarriers and 2.0x105cells/mL was successful. A maximum cell yield of 1.2x106cells/mL was obtained in stirred-tank microcarrier batch culture while cell numbers in the Wave®Bioreactor could not be determined accurately due to the fast sedimentation of microcarriers under non-rocking conditions required for sampling. Detailed off-line analysis was carried out to understand the behaviour of the virus-host cell system in both cultivation systems. Metabolic profiles for glucose, lactate, glutamine, and ammonium showed slight differences for both systems. E-FL cell growth was on the same level in stirred-tank and Wave®Bioreactor with a higher volumetric cell yield compared to roller bottles. Propagation of MEV, which can only replicate efficiently in mitotic cells, was characterized in the Wave®Bioreactor using a multiple harvest strategy. Maximum virus titres of 106.6to 106.8TCID50/mL were obtained, which corresponds to an increase in virus yield by a factor of about 10 compared to cultivations in roller bottles. As a consequence, a single Wave®Bioreactor cultivation of appropriate scale can replace hundreds of roller bottles. Thus, the Wave®Bioreactor proved to be a suitable system for large-scale production of an inactivated MEV vaccine.