Serum-free microcarrier based production of replication deficient Influenza vaccine candidate virus lacking NS1 using Vero cells
| Autor: | Say Kong Ng, Allen Chen, Mylene L Yan, Elisabeth Roethl, Christian Dietzsch, Swan Li Poh |
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| Jazyk: | angličtina |
| Předmět: |
Microcarrier
Virus Cultivation Influenza vaccine lcsh:Biotechnology viruses NS1 Bioreactor Viral Nonstructural Proteins Biology Vaccines Attenuated Virus Replication medicine.disease_cause Culture Media Serum-Free Virus Microbiology Bioreactors Influenza A Virus H1N1 Subtype lcsh:TP248.13-248.65 Chlorocebus aethiops Influenza A virus medicine Animals Live attenuated influenza vaccine Microscopy Phase-Contrast Vero Cells Duck embryo vaccine Attenuated vaccine Virology Influenza Vero Influenza Vaccines Inactivated vaccine Vero cell Research Article Biotechnology |
| Zdroj: | BMC Biotechnology, Vol 11, Iss 1, p 81 (2011) BMC Biotechnology |
| ISSN: | 1472-6750 |
| DOI: | 10.1186/1472-6750-11-81 |
| Popis: | Background Influenza virus is a major health concern that has huge impacts on the human society, and vaccination remains as one of the most effective ways to mitigate this disease. Comparing the two types of commercially available Influenza vaccine, the live attenuated virus vaccine is more cross-reactive and easier to administer than the traditional inactivated vaccines. One promising live attenuated Influenza vaccine that has completed Phase I clinical trial is deltaFLU, a deletion mutant lacking the viral Nonstructural Protein 1 (NS1) gene. As a consequence of this gene deletion, this mutant virus can only propagate effectively in cells with a deficient interferon-mediated antiviral response. To demonstrate the manufacturability of this vaccine candidate, a batch bioreactor production process using adherent Vero cells on microcarriers in commercially available animal-component free, serum-free media is described. Results Five commercially available animal-component free, serum-free media (SFM) were evaluated for growth of Vero cells in agitated Cytodex 1 spinner flask microcarrier cultures. EX-CELL Vero SFM achieved the highest cell concentration of 2.6 × 10^6 cells/ml, whereas other SFM achieved about 1.2 × 10^6 cells/ml. Time points for infection between the late exponential and stationary phases of cell growth had no significant effect in the final virus titres. A virus yield of 7.6 Log10 TCID50/ml was achieved using trypsin concentration of 10 μg/ml and MOI of 0.001. The Influenza vaccine production process was scaled up to a 3 liter controlled stirred tank bioreactor to achieve a cell density of 2.7 × 10^6 cells/ml and virus titre of 8.3 Log10 TCID50/ml. Finally, the bioreactor system was tested for the production of the corresponding wild type H1N1 Influenza virus, which is conventionally used in the production of inactivated vaccine. High virus titres of up to 10 Log10 TCID50/ml were achieved. Conclusions We describe for the first time the production of Influenza viruses using Vero cells in commercially available animal-component free, serum-free medium. This work can be used as a basis for efficient production of attenuated as well as wild type Influenza virus for research and vaccine production. |
| Databáze: | OpenAIRE |
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