Establishment of a mink enteritis vaccine production process in stirred-tank reactor and Wave Bioreactor microcarrier culture in 1-10 L scale.
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 1 microcarriers and 2.0 x 10(5)cells/mL was successful. A maximum cell yield of 1.2 x 10(6)cells/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 10(6.6) to 10(6.8) TCID(50)/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.
Hundt
B
B
Impfstoffwerk Dessau-Tornau GmbH, PF 400214, 06855 Rosslau, Germany. boris.hundt@idt-direct.de
Best
C
C
Schlawin
N
N
Kassner
H
H
Genzel
Y
Y
Reichl
U
U
D000818:Animals; D019149:Bioreactors / Q000821:virology*; D002415:Cats; D018929:Cell Culture Techniques / Q000379:methods; D004622:Embryo, Mammalian; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism / Q000821:virology*; D008907:Mink; D053490:Mink enteritis virus / Q000254:growth & development / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D015164:Vaccines, Inactivated / Q000096:biosynthesis; D014765:Viral Vaccines / Q000096:biosynthesis*; D014776:Virus Cultivation / Q000379:methods; D014779:Virus Replication
D015164:Vaccines, Inactivated; D014765:Viral Vaccines