1. Production of Hepatitis A Virus using TideCell Bioreactor System www.bioreactorsciences.com A case study using GL37cell line

2. 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.

3. (R1) Cells are highly sensitive to shear stress during post-infection period TideCell: Extremely low shear stress environment in TideCell enables cells not easily to detach after infection and increase the productivity. Microcarrier system: shear stress is relatively higher and cells tend to detach during the post infection period.

4. (R2) Virus doesn’t cause Cytopathic effect (CPE). Entire production process is long and requires exchanging culture medium continuously. TideCell: Low shear stress and easy medium exchange allow a long cell culture cycle in TideCell. Microcarrier system: can be done with perfusion mode

5. (R3) Virus is non-secreted and require cell harvest and disruption to harvest the cells TideCell:Cells immobilized in matrix vessel which is separated from mixing media vessel and can be separated for cell harvest process Microcarrier system requires to process the entire tank to separate the beads from media. Other packed bed system cannot remove carriers for cell harvest.

6. (R4) Virus productivity is sensitive to nutrient supply and cell growth rate. Higher cell growth rate results lower virus productivity. TideCell won’t cause cell over growth. Easy to control cell growth by reducing tidal frequency and limiting nutrient supply. A simple starving procedure enhances protein or virus expression Microcarrier and other packed bed systems can only do nutrient control by controlling perfusion rate. Not easy to do. Roller bottle: no nutrient control available

7. Final results TideCell: 0.5 mg/L bioreactor Roller bottle:0.5 mg/L Microcarrier system: 0 mg/L. Fail to culture cells during post-infection period due to serious cell detachment problems.

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