Production of Hog Cholera Virus using TideCell Bioreactor System A production case study using ST cell line

Comparison study  Due to significant difference in characteristics of virus strain and host cell line, the optimal process of virus production can be significantly different.  In the following slides are requirement of conditions for this specific case of study, on which comparison of TideCell/BelloCell system and other commonly used systems are based.

(R1) High cell density is required to achieve high titer, and reduce medium consumption.  TideCell: High surface area provided increase cell density up to 10 folds and save culture medium up to 90%.  Roller bottle: low cell density due to limitation of surface area  Agitated Microcarrier system: the bead density is limited  Other Fixed bed system: Yes, but the scale is limited.

(R2) Virus won’t cause Cytopathic effect, and a long term harvest is achievable. What be done?  Low shear stress, less detachment and easy medium exchange allows a long harvest cycle in TideCell. over 28 harvest cycles are achieved in TideCell.  Roller bottles: Only up to 4 cycles  Agitated Microcarrier system: Shear stress is higher. Cells tend to detach during post-infection period  Other fixed bed system: Shear stress is higher. Cells tend to detach during post-infection period.

(R3) Require to sample carrier and observe to determine the infection time by morphology and cell count  All systems are capable of sampling and observation except packed bed bioreactor systems other than TideCell system

(R4) Virus is highly unstable: sensitive to temperature. Daily semi-harvest is required.  Easy to do medium exchange and harvesting without interrupting cell culture process.  Roller bottle: Each bottle has to be removed from culture.  Microcarrier system: System has to stop and allow microcarrier to settle.  Other Fixed bed system: liquid level cannot be lowered than the working volume. Medium exchange is limited

(R5) Require to use Low serum or zero serum to reduce side effect to pigs  Low shear stress culture allow low serum or serum- free culture environment without causing cell detach and loss.  Roller bottle: Cells tend to detach in serum-free culture.  Microcarrier system: Cells tend to detach in low serum or serum-free culture environment  Other Fixed bed system: Depending on the agitation and recirculation speed during culture.

Process profiles

Results  Increase titer from 0.2 mil RIB (roller bottle) to 6 mil. RIB a total 30 fold increase.  Increase harvest cycle from 4 (roller bottle) to 28.  Save culture medium 1000 L per batch  No data on other systems

Summary of result and comparison

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