1. Continuous Purity™ Marc Bisschops Tarpon Biosystems Inc. Integrated Continuous Biomanufacturing Castelldefels – Spain October 20 – 24, 2013 Technological, Regulatory and Validation Considerations for single-use continuous downstream processing

2. Continuous Manufacturing Photo: courtesy Martien Tazelaar (taas.it)

3. Continuous Manufacturing Generalized Mass Balance for a single phase: Why are transient processes so hard to design and control? Dispersion – Convection + Reaction = Accumulation Gradient in SpaceTime dependent

4. Continuous Manufacturing Generalized Mass Balance for a single phase: Why are continuous processes easier to design and control? Dispersion – Convection + Reaction = Steady State Gradient in Space

5. Continuous Manufacturing Over the past 40 years, the vast majority of accidents in chemical industries happened during non-routine manufacturing operations (mainly during start-up). W. Bridges and T. Clark (2011) Chemical catastrophe in 2008 after anomalies during a start-up of a chemical facility in West Virginia. This resulted in a runaway chemical reaction, causing a pressure vessel to explode. The accident killed 2 employees of the company and eight people were injured. (US Chemical Safety Board Report 2008-08-I-WV, Jan.2011)

6. ... or Batch Manufacturing Biopharmaceutical industries: Product quality is directly related to process control (“The Process is the Product”) Batch processes are – almost by definition – transient processes So, if batch processes are more difficult to control, and if biopharmaceutical product quality is so tightly related to process control... then shouldn’t we at least consider Continuous Biomanufacturing?

7. Regulatory Aspects Batch Definition: No specific regulations or guidance for in continuous manufacturing (can be based on time or materials supply) Should be based on assurance of consistent product quality (e.g. equipment cycles or material properties) Nothing in regulations or guidance prohibiting continuous manufacturing S. Chatterjee, FDA Perspective on Continuous Manufacturing, IFPAC Meeting, Jan 2012

8. Continuous Manufacturing Translating batch to continuous: Challenges of continuous DSP may be less than continuous USP (in terms of product quality control) USPDSP Longer processing times increases chance of product heterogeneity Shorter residence times decreases chance of product degredation or contamination Impacts micro-environment & chemistry of cells Utilizes the same fundamental chemistry as batch

9. Simplifying the PFD MAb manufacturnig platform, presented by Wolfgang Berthold (2008)

10. Simplifying the PFD

11. Capital Utilization Continuous processing: Saves time in suite by 50 – 70% Minimize footprints of some of the large unit operations All unit operations sized by volume (instead of mass of protein) Batch ProcessingContinuous Processing

12. Continuous AND Disposable

13. Technological Solutions Compatibility chart for common DSP Unit Operations in Continuous and Single-Use format Process StepContinuousSingle-Use Clarification: Centrifugation Clarification: Depth Filtration Chromatography: Capture Virus inactivation Chromatography: Polishing (AEX) Chromatography: Polishing (CEX) Ultrafiltration Virus filtration      /  /        

14. Continuous Disposable Chromatography Tarpon Biosystems’ BioSMB ® Key features: Multicolumn chromatography: continuous and countercurrent process Higher specific productivity Single use valve cassette FeatureBenefit Countercurrent processImproved resin capacity utilization High specific productivityReduced resin inventory BioSMB Valve CassetteFully disposable flow path Configuration flexibility

15. BioSMB ® Process Development System

16. System Design & Segregation of Fluids pH UV C pH UV C pH UV C pH UV C Batch Chromatography SkidContinuous Chromatography Skid Segregated: one fluid throughout batch Shared: multiple process solutions throughout batch

17. System Design & Segregation of Fluids Segregation of fluids in batch and continuous systems: Continuous systems have an inherently better segregation of process solutions Process StepBatchContinuous Buffer selection valves SharedNA Pump(s) SharedSegregated Sensors (inlet) SharedSegregated Column bypass valves SharedNA Integrated valve system NAPartly shared Sensors (outlet) SharedSegregated Outlet selection valves SharedNA

18. System Design & Sensors Sensors are dedicated to an individual outlet: More sensors provide more information on the process Will be operated in a more narrow range and can therefore be selected to meet higher accuracy (e.g. flow path in UV flow cells) Can be selected to meet the specifics of that particular outlet (e.g. UV wavelengths)

19. FMEA Risk Ranking (General) Continuous versus Batch SeverityImpact on CQA is identical due to nature of the process Consequence may, however, not affect entire batch but only small increment (small repetitive cycles) OccurrenceMore complex equipment may lead to (perception of) higher probability of failure DetectionContinuous process will immediately detect deviations whereas batch process may only detect afterwards Overall rankingContinuous process might rank better than batch process

20. FMEA Risk Ranking (Abbreviated) SeverityOccurrenceDetection Column Failure Direct impact on CQA Very low probability (1) Immediate Pump Failure Potential impact on CQA Low probabilityImmediate Valve Failure Potential impact on CQA Very low probability (2) Immediate Detector Failure No impact on CQA Low ProbabilityImmediate (1)Probability of column failure can be significantly reduced by using smaller diameters, prepacked & pretested columns (2)Probability of valve failure can be significantly reduced by implementing valve integrity tests before running a batch

21. Experience with BioSMB – Valve Integrity Mean time to failure of disposable valve technology: Note: Main causes of failure for diaphragm valves are related to the diaphragm, particularly in combination with a steam cycle. Disposable components are generally not steamed. BioSMB Valve Technology Basis of DesignBased on traditional diaphragm valve technologies 10 5 – 10 6 cycles TestedRapid cycling tests of BioSMB valve cassette (all valves) 10 4 cycles Intended useIntended use of BioSMB cassette corresponds to column life time 10 2 cycles M. Bridge on PharmTech.com, June 2011

22. Experience with BioSMB – Consistency Rapid cycling provides repetitive response of sensors Deviations can be immediately recognized Four column BioSMB process for capture of Monoclonal Antibodies using Protein A affinity chromatography

23. Experience with BioSMB – Dynamics Start-up and shut-down cycles: Dedicated methods for accellerated start-up and shut-down cycles can be used Product concentration may vary, impurity profile remains constant (only effect is dilution) Recovering from process upsets: Response to step changes is very fast (less than one process cycle)

24. Overall Process Lay-out Integrated continuous biomanufacturing process: Large intermediate product hold tanks are eliminated Small surge bags between unit operations may address flow control and cyclic behaviour Mitigation of potential process hick-up downstream: emergency surge bag FeatureBenefit Controlled residence timesProduct quality control Shorter processing timeProduct quality control Smaller process equipmentFavors disposable bioprocessing technologies

25. Common Reasons for Batch Failure Contamination: Disposable components Minimizing residence times Segregation of fluids Operator Error: Automation Training Equipment Failure: Automation Testing protocols E. Langer, BioProcess International, September 2008

26. Common Reasons for Batch Failure Over the past five years, average batch failures have been reduced significantly (appr 50% decline). Mean causes: Improved process design (including QbD) Improved process monitoring (including PAT) Operator Training E. Langer, Pharmaceutical Manufacturing, June 2012

27. Conclusions Although more complex, continuous process technologies are likely to comply to cGMP requirements as well as batch alternatives: Better segregation of process solutions and shorter processing times minimizes risk of contamination Immediate feed back & rapid staedy state cycling limits consequence of potential process upsets Continuous processing fits naturally with PAT initiatives Continuous processing and disposable processing are natural partners

28. It requires courage to take hurdles It may well be that the first implementations of continuous processes may not deliver the full promise Redefine validation strategies Redefine quality systems Beat organizational hurdles... That should not keep us from pursuing promising technologies J.L Bower and C.M. Christensen, Harvard Business Review, Jan/Feb 1995

29. Acknowledgements Tom Ransohoff (BPTC) Lynne Frick (Tarpon Biosystems) All companies exploring continuous biomanufacturing "People are moving now to continuous manufacturing and really much more high tech modern ways and it doesn't fit the way good manufacturing practice has been thought about over the years," Woodcock said. "We have to forcibly make sure we allow the better to come about." Janet Goodwin Head of FDA Pharmaceutical Division Reuters, October 10, 2013