Please use this identifier to cite or link to this item: https://doi.org/10.1186/1472-6750-11-81
Title: Serum-free microcarrier based production of replication deficient Influenza vaccine candidate virus lacking NS1 using Vero cells
Authors: Chen, A
Poh, S.L
Dietzsch, C
Roethl, E
Yan, M.L
Ng, S.K 
Keywords: Antiviral response
Batch bioreactors
Bioreactor system
Cell concentrations
Cell density
Clinical trial
Deletion mutants
Efficient production
Gene deletion
Health concerns
Human society
Influenza
Influenza vaccines
Influenza virus
Manufacturability
Microcarrier
Microcarrier culture
Microcarriers
Nonstructural proteins
NS1
Phase I
Production process
Serum-free media
Serum-free medium
Spinner flasks
Stationary phasis
Stirred tank bioreactors
Time points
Vaccine production
Vero
Vero cells
Wild types
Animals
Bioconversion
Bioreactors
Body fluids
Cell culture
Cells
Engineering research
Genes
Growth kinetics
Production engineering
Vaccines
Viruses
Diseases
cytodex 1
drug carrier
nonstructural protein 1
trypsin
unclassified drug
influenza vaccine
INS1 protein, influenza virus
live vaccine
virus protein
article
batch reactor
cell adhesion
cell culture
cell density
cell growth
cell viability
hemagglutination inhibition
influenza A
Influenza virus A
Influenza virus A H1N1
nonhuman
stirred reactor
Vero cell
virogenesis
virus replication
animal
bioreactor
biosynthesis
Cercopithecus
comparative study
culture medium
genetics
growth, development and aging
Influenza virus A H1N1
instrumentation
metabolism
methodology
phase contrast microscopy
physiology
Vero cell
virology
virus culture
Animalia
Orthomyxoviridae
Animals
Bioreactors
Cercopithecus aethiops
Culture Media, Serum-Free
Influenza A Virus, H1N1 Subtype
Influenza Vaccines
Microscopy, Phase-Contrast
Vaccines, Attenuated
Vero Cells
Viral Nonstructural Proteins
Virus Cultivation
Virus Replication
Issue Date: 2011
Citation: Chen, A, Poh, S.L, Dietzsch, C, Roethl, E, Yan, M.L, Ng, S.K (2011). Serum-free microcarrier based production of replication deficient Influenza vaccine candidate virus lacking NS1 using Vero cells. BMC Biotechnology 11 : 81. ScholarBank@NUS Repository. https://doi.org/10.1186/1472-6750-11-81
Rights: Attribution 4.0 International
Abstract: 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 Log10TCID50/ml was achieved using trypsin concentration of 10 ?g/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 Log10TCID50/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 Log10TCID50/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. © 2011 Chen et al; licensee BioMed Central Ltd.
Source Title: BMC Biotechnology
URI: https://scholarbank.nus.edu.sg/handle/10635/181628
ISSN: 14726750
DOI: 10.1186/1472-6750-11-81
Rights: Attribution 4.0 International
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