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A production case study using MDCK cell line

Production of Avian Influenza H5N1 Virus Virus using TideCell Bioreactor System www.bioreactorsciences.com. A production case study using MDCK cell line. Comparison study.

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A production case study using MDCK cell line

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  1. Production of Avian Influenza H5N1 VirusVirus using TideCell Bioreactor Systemwww.bioreactorsciences.com A production case study using MDCK cell 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) 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%. • Microcarrier system: bead density is limited • Other packed bed system: scale is limited

  4. (R2) Cell attachment efficiency is low due to long term trypsinization • TideCell: A relative static seeding method enable high attachment rate even the cells have been over-trypsinized • Microcarrier system:. An agitation environment increases the difficulties of cell attachment. The cell attachment efficiency becomes lower when the scale is increased. • Other fixed bed system: An agitation environment increases the difficulties of cell attachment.

  5. (R3) Cells tend to detach after infection due to the adding of trypsin • TideCell: Extremely low shear stress provided low shear stress culture environment by TideCell allows cells not easy to detach after infection • Other systems other than Roller bottle require agitation which may cause cell detachment after infection.

  6. (R4) Require to contain low impurities including DNA and low host cell protein in the harvest • TideCell: Low shear stress results in less cell disruption and less impurities in the harvest. • Microcarrier system: An agitated environment causes cell disruption and release of DNA and host cell protein. • Other fixed bed system: same as above.

  7. (R5) Require to refresh culture medium before infection • TideCell: Cells are immobilized in the matrix vessel which is separated from mixing vessel containing culture medium. Medium could be exchanged directly and slowly without losing cells and causing temperature shock. • Microcarrier system. Medium can only be partially replaced by settling the carriers. Nutrient may not be sufficient to support entire post-infection period. • Other packed-bed system: medium has to be filled completely and a temperature shock might be ocurred especially in a large scale system.

  8. Results of cell growth in BelloCell

  9. Virus HA Titer in BelloCell

  10. Process Flow Diagram

  11. Cell Growth before Infection

  12. Morphology after Infection

  13. HA profile

  14. SDS-page

  15. Virus production in variour media

  16. PFU profiles

  17. HA profiles

  18. Summary of Results

  19. Thank you for watching Please visit us @ www.bioreactorsciences.com or www.cescobioproducts.com

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