Abstract
MedImmune Vaccines has engineered a live, attenuated chimeric virus that could prevent infections caused by parainfluenza virus type 3 (PIV3) and respiratory syncytial virus (RSV), causative agents of acute respiratory diseases in infants and young children. The work here details the development of a serum-free Vero cell culture production platform for this virus vaccine candidate. Efforts to identify critical process parameters and optimize culture conditions increased infectious virus titers by approximately 2 log10 TCID50/ml over the original serum-free process. In particular, the addition of a chemically defined lipid concentrate to the pre-infection medium along with the shift to a lower post-infection cultivation temperature increased virus titers by almost 100-fold. This improved serum-free process achieved comparable virus titers to the serum-supplemented process, and demonstrated consistent results upon scale-up: Vero cultures in roller bottles, spinner flasks and bioreactors reproducibly generated maximum infectious virus titers of 8 log10 TCID50/ml.
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Abbreviations
- CDLC:
-
Chemically defined lipid concentrate
- F:
-
Fusion
- FBS:
-
Fetal bovine serum
- HN:
-
Hemagglutinin–neuraminidase
- hPIV3:
-
Human parainfluenza virus type 3
- MEDI-534:
-
Chimeric human parainfluenza virus type 3/respiratory syncytial virus
- MOI:
-
Multiplicity of infection
- PIV3:
-
Parainfluenza virus type 3
- RB:
-
Roller bottle
- RSV:
-
Respiratory syncytial virus
- SFM:
-
Serum-free medium
- TCID50 :
-
50% Tissue culture infective dose
- Vero:
-
African green monkey kidney
- VP-SFM:
-
Virus production serum-free medium
- WME:
-
Williams’ Medium E
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Acknowledgements
We thank Richard R. Spaete, Roderick S. Tang, Mia MacPhail, Jeanne H. Schickli, Jeanne M. Guzzetta, Jasmine Kaur, Elizabeth Stillman, and Aurelia Haller for generating the original MEDI-534 vectors by plasmid rescue.
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Yuk, I.H., Lin, G.B., Ju, H. et al. A serum-free Vero production platform for a chimeric virus vaccine candidate. Cytotechnology 51, 183–192 (2006). https://doi.org/10.1007/s10616-006-9030-7
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DOI: https://doi.org/10.1007/s10616-006-9030-7