Development and optimisation of a procedure for the production of Parapoxvirus ovis by large-scale microcarrier cell culture in a non-animal, non-human and non-plant-derived medium
| Autor: | D. Paulsen, Helga Rübsamen-Waigmann, Uwe Langer, Torsten Minuth, Heiner Apeler, H. J. Henzler, Hans-Dietrich Horlein, Michael Pohlscheidt, Udo Reichl, Berthold Bödeker |
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| Rok vydání: | 2007 |
| Předmět: |
Cell Survival
Cell Cell Count Mice Transgenic Kidney Mice Multiplicity of infection medicine Animals Poxviridae Trypsin Cells Cultured Parapoxvirus Chromatography General Veterinary General Immunology and Microbiology biology Public Health Environmental and Occupational Health Kidney metabolism Microcarrier biology.organism_classification Hepatitis B Virology Culture Media Microscopy Electron Infectious Diseases medicine.anatomical_structure Vaccines Inactivated Cell culture DNA Viral Batch processing Molecular Medicine Cattle |
| Zdroj: | Vaccine. 26(12) |
| ISSN: | 0264-410X |
| Popis: | For the production of a chemically inactivated Parapoxvirus ovis (PPVO), an adherent bovine kidney cell line was cultivated on Cytodex-3 microcarriers in suspension culture. The inactivated and purified virus particles have shown immune modulatory activity in several animal models. PPVO was produced by a biphasic batch process at the 3.5 and 10 L scale. Aeration was realised by bubble-free membrane oxygenation via a tube stator with a central two-blade anchor impeller. In order to increase efficiency, process robustness and safety, the established process was optimised. The cell line was adapted to a protein-free medium (except recombinant insulin) in order to increase biosafety. A scale up to a 50 L pilot plant with direct cell expansion was performed successfully. In parallel, the biphasic batch process was optimised with special emphasis on different operating conditions (cell number, Multiplicity of Infection (MOI), etc.) and process management (fed-batch, dialysis, etc.). The quality and concentration of the purified virus particles was assessed by quantitative electron microscopy, residual host cell protein and DNA-content and, finally, biologic activity in a transgenic mouse model. This integrated approach led to a new, safe, robust and highly productive large-scale production process, called "Volume-Expanded-Fed" Batch with cell densities up to 6-7e06 cells/mL. By subsequent dilution of infected cells into the next process scale, an increase in total productivity by a factor of 40 (related to an established biphasic batch process) was achieved. |
| Databáze: | OpenAIRE |
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