Production of Avian Influenza H5N1 Virus Virus using

TideCell Bioreactor Systemwww.bioreactorsciences.com

A production case study using MDCK 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%.

Microcarrier system: bead density is limited

Other packed bed system: scale is limited

(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.

(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.

(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.

(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.

Results of cell growth in BelloCell

Virus HA Titer in BelloCell

Process Flow Diagram

Cell Growth before Infection

Morphology after Infection

HA profile

SDS-page

Virus production in variour media

PFU profiles

HA profiles

Summary of Results

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