Elsevier

Vaccine

Volume 32, Issue 24, 19 May 2014, Pages 2782-2788
Vaccine

Improved poliovirus d-antigen yields by application of different Vero cell cultivation methods

https://doi.org/10.1016/j.vaccine.2014.02.022Get rights and content
Under a Creative Commons license
open access

Highlights

  • Vero cells were grown in batch, semi-batch, perfusion and recirculation strategies.

  • At high cell densities (to 5 × 106 cells mL−1) cells were infected with poliovirus.

  • Increased cell densities allowed 3 fold increase in d-antigen yield.

  • Cell specific d-antigen yields were lower at higher cell densities.

  • The semi-batch cultivation strategy is most promising for optimization.

Abstract

Vero cells were grown adherent to microcarriers (Cytodex 1; 3 g L−1) using animal component free media in stirred-tank type bioreactors. Different strategies for media refreshment, daily media replacement (semi-batch), continuous media replacement (perfusion) and recirculation of media, were compared with batch cultivation. Cell densities increased using a feed strategy from 1 × 106 cells mL−1 during batch cultivation to 1.8, 2.7 and 5.0 × 106 cells mL−1 during semi-batch, perfusion and recirculation, respectively.

The effects of these different cell culture strategies on subsequent poliovirus production were investigated. Increased cell densities allowed up to 3 times higher d-antigen levels when compared with that obtained from batch-wise Vero cell culture. However, the cell specific d-antigen production was lower when cells were infected at higher cell densities. This cell density effect is in good agreement with observations for different cell lines and virus types. From the evaluated alternative culture methods, application of a semi-batch mode of operations allowed the highest cell specific d-antigen production.

The increased product yields that can easily be reached using these higher cell density cultivation methods, showed the possibility for better use of bioreactor capacity for the manufacturing of polio vaccines to ultimately reduce vaccine cost per dose. Further, the use of animal-component-free cell- and virus culture media shows opportunities for modernization of human viral vaccine manufacturing.

Keywords

Microcarriers
Perfusion
Feed
Recirculation
Batch
Adherent

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