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Optimization of T cell expansion in a perfusion bioreactor

Optimization of T cell expansion in a perfusion bioreactor. Clive Glover PhD Product Leader, Cell Bioprocessing. Perspective. Scaling UP?. Scaling OUT?. “Home”. Wikipedia.com. 123RF.com. ?. Industry. What does this even look like?. 123RF.com. Chimeric Antigen Receptor T cells- CARTs.

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Optimization of T cell expansion in a perfusion bioreactor

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  1. Optimization of T cell expansion in a perfusion bioreactor Clive Glover PhD Product Leader, Cell Bioprocessing

  2. Perspective Scaling UP? Scaling OUT? “Home” Wikipedia.com 123RF.com ? Industry What does thiseven look like? 123RF.com

  3. Chimeric Antigen Receptor T cells- CARTs Antibody variable region TC TC TC TH TH TH T cell Receptor intracellular signalling component

  4. Clinical Trials Results

  5. CART – Chimeric Antigen Receptor T cells + T cells Lentiviral – expressing Chimeric Antigen Receptor Cell Infusion into Patient Cell Harvest & Concentration CAR T cells

  6. Typical cell dose = 1x108/kg 20 kg patient = 2 x 109 cells 100 kg patient = 1 x 1010 cells

  7. Factory Scale Cell Collection Cell Collection Cell Collection Cell Collection Cell Collection Cell Collection Cell Collection Cell Collection Cell Collection Cell Collection Cell Collection Cell Collection Cell Collection • 5000 patients • Process time = 10 days • Number of patients in parallel = 140 Cell Separation Cell Separation Cell Separation Cell Separation Cell Separation Cell Separation Cell Separation Cell Separation Cell Separation Cell Separation Cell Separation Cell Separation Cell Separation Cell Infusion into Patient Cell Infusion into Patient Cell Infusion into Patient Cell Infusion into Patient Cell Infusion into Patient Cell Infusion into Patient Cell Infusion into Patient Cell Infusion into Patient Cell Infusion into Patient Cell Infusion into Patient Cell Infusion into Patient Cell Infusion into Patient Cell Infusion into Patient Cell Selection Cell Selection Cell Selection Cell Selection Cell Selection Cell Selection Cell Selection Cell Selection Cell Selection Cell Selection Cell Selection Cell Selection Cell Selection Cell Harvest & Concentration Cell Harvest & Concentration Cell Harvest & Concentration Cell Harvest & Concentration Cell Harvest & Concentration Cell Harvest & Concentration Cell Harvest & Concentration Cell Harvest & Concentration Cell Harvest & Concentration Cell Harvest & Concentration Cell Harvest & Concentration Cell Harvest & Concentration Cell Harvest & Concentration Cell Activation & Expansion Cell Activation & Expansion Cell Activation & Expansion Cell Activation & Expansion Cell Activation & Expansion Cell Activation & Expansion Cell Activation & Expansion Cell Activation & Expansion Cell Activation & Expansion Cell Activation & Expansion Cell Activation & Expansion Cell Activation & Expansion

  8. Key Requirements of Cell Therapy Manufacturing Processes • Scalable. • Sample contained in 1 vessel • Easy to scale out to make most efficient use of manufacturing space • Automatableto minimize the chance of human error • Single Use and Traceableto eliminate cross contamination with other patient cells • Closed system to eliminate chance of contamination with adventitious agents due to handling • Robust and Compliant. To ensure consistency of product and satisfaction of regulatory requirements

  9. WAVE 2/10Closed. Automated. Single-use

  10. Growth kinetics Total Cell No. Day of Culture

  11. Optimization Studies • Objective: Maximize the expansion of viable T cells in a 10 day period 2,6 2,2 2,9 Angle Rocking Speed 10,9 10,2 10,6 18,2 18,9 18,6

  12. Experimental Design Day of culture 5 6 7 8 9 10 0 1 2 3 4 Culture to 1L Perfuse 500mls Perfuse 750mls • Daily monitoring of: • Cell proliferation/viability • Glucose/Lactate/Ammonia Perfuse 1L

  13. Experimental Design QC analysis 5 10 0 6 7 8 9 1 2 3 4 • Phenotype monitoring of: • CD4/CD8 ratio • CD27/CD28 expression to assess differentiation state • CD57 expression to assess the presence of senescent cells • CD62L expression to assess migratory ability

  14. Results • No significant effects of angle or rpm on cell health

  15. Results • Significant effect of rocking speed on cell expansion

  16. Optimization Fold expansion sum Optimized speed and angle: 15.02 rpm, 5.625 º

  17. Optimization Cell count (106/ mL) Day

  18. Summary • Autologous cellular immunotherapies have unique scalability requirements • WAVE systems provide robust and reliable expansion of functional T cells • 10% increase in cell yield using optimized bioreactor settings • Higher cell densities and a closed and automated system make them ideal for therapeutic use

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