Bradley E Kosiba, PhD, CPIP BK Collaborative, LLC ECI Integrated Continuous Biomanufacturing

1. Facility Drivers for Housing Start-to-Finish Continuous Bioprocessing.Disruptive changes in scale & operational expectations vs. traditional batch operations Bradley E Kosiba, PhD, CPIP BK Collaborative, LLC ECI Integrated Continuous Biomanufacturing Clinical and Commercial Facility Design for Continuous Biomanufacturing October 23, 2013

2. Scope of Disruption • Facility Drivers • Process Scale • Process Duration • Process Integration • Points of Disruption • Process Development • Clinical Production • Manufacturing • A Few Technology Gaps • Miniaturization of Biotech

3. ScaleCompare to 13,000L Fed Batch BioRx • Production Bioreactor Harvests • 3 X 13,000L fed batch per week = 1 X 3,700L Perfusion @1.5V/day At similar titers this leads to… • Seeds per Quarter (3 months) • 36 batch vs. 1 perfusion • Downstream piping flows (approx.) • Batch – 50 LPM transfer/250 LPM UF retentate • Continuous – 5 LPM harvest, 0.5 LPM polishing, 0.05 LPM final DS • Tank volume for Unit Operations (UOs) • Batch: 1,000 – 20,000L Process Tanks.…….. 200L final DS (3/week) • Continuous: 20 – 500L Surge Tanks…………. 200L final DS (3/week)

4. Integration The Slippery Slope to Continuous BioProcessing • Demise of “hold and test” steps • Directly replace offline with PAT • Rise of QbD • Increases predictability • Reduce offline confirmation testing • Move to mini-batch chrom • Limit column size • Upgrade to continuous mini-batch operation • Combine UOs for efficiency • Pairing flow-through chrom with next batch step • Full automation reduces human error • Automate transitions between unit ops/cleaning/prep

5. Duration • Perfusion duration to 3 months (or more?) • Fully integrated downstream = same duration • Equipment Sanitation • Minibatch processes allow periodic cleaning • Challenge: maintain sanitation of remaining flow paths • Equipment/Sensor/Separation Media Durability • Change out/recalibration needed? • SU disposables capable of 3+ month duration? • Product/Batch Uniformity • Steady state perfusion supports glycosylation uniformity • Downstream process uniformity eliminates final batch mix?

6. Scope of Disruption • Facility Drivers • Process Scale • Process Duration • Process Integration • Points of Disruption • Process Development • Clinical Production • Manufacturing • A Few Technology Gaps • Miniaturization of Biotech

7. Process Development • Scale • Scaled down continuous UO availability? • 50L BioRx = 0.5 - 50 ml/min downstream • Integration • Automation of Lab-scale Integrated Continuous Processes • User friendly • Whole process • Multi vendor UO tolerant? • Duration • Steady State Development Platform Benefits • Iterative automated DOE design space mapping over long runs • Final Drug Substance (DS) or Product (DP) quality readout • And Challenges • Sanitation of small, non-GMP flow paths • Maintaining continuous buffer supplies

8. Clinical Production • Scale • Clinical batches made at full manufacturing “scale” (not duration) • Duration • Clinical Batches from multiple short duration runs • Clinical Batches collected sequentially from one long duration run • Combinations • Integration • Requires full, integrated process & automation program • Stable Steady-State processes • Could facilitate clinical experience at non-center point conditions • Planned excursions within design space during clinical runs • Collect “batches” under different parameter regimes

9. Commercial Production • Scale • 1x 3,700 L culture vs. 3x 13,000L cultures • 0.05 – 5 LPM vs. 50 – 250 LPM • Integration • Full automation of Continuous process • Automated transitions between process, cleaning and prep cycles • No holds or stops between Unit Operations • Duration • Process Validation • Relevant experience from continuous clean utility systems (WFI, CS, CAP) • Relies heavily on robust QbD package to gain confidence before PV runs • Lack of routine human intervention reduces uncertainty • True Risk-based batch sizing • vs. Risk + Capital cost based sizing

10. Scope of Disruption • Facility Drivers • Process Scale • Process Duration • Process Integration • Points of Disruption • Process Development • Clinical Production • Manufacturing • A Few Technology Gaps • Miniaturization of Biotech

11. Technology Gaps Continuous single-pass buffer exchange (GMP) • Multi-stage UF/dilution • brute force engineering • Multi-stage 1-pass UF concentration exists in food industry • Countercurrent 1-pass UF • Similar to renal dialysis • Probably multi-stage pumping • Continuous Size Exclusion Chromatography desalting • Simple Vo vs. Vi separation • Uses current multi-column technology • Dilutes product (requires 1-pass UF concentration for final DS) • Available today

12. Technology Gaps Properly Scaled Unit Operations • Current sizing equal to batch sized UO’s • Integrated Continuous BioProcessing at smaller scale! • Even further scale-down for bench-scale development • Sanitation at smaller scale for long runs a challenge! • Disposable fluid paths may helpnon-minibatch UO’s

13. The Miniaturization of Biotech • Early Biotech • 100’s L BioRx and Downstream process • Tanks 10’s - 100’s L • 1 – 5 LPM • Current Biotech • 6 X 10,000+ L BioRx and Downstream processes • Tanks 10,000’s L • 50 – 250 LPM • Miniature Integrated Continuous Bioprocess • 1-2 X 1,000’s L BioRx • Tanks 10’s – 100’s L • 0.05 – 5 LPM

14. Summary • Scale, Integration, Duration • Process Development, Clinical Production, Commercial Prod’n • Technology Gaps • Miniaturization of Biotech (Facilities)

15. Acknowledgements • Rick Lawless, Associate Director, NCSU BTEC • NCSU Chem Engineering CBioP Senior Project Team 2012-13