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| www.genzyme.com Jade (with her mother) Fabry disease USA UPSTREAM DEVELOPMENT OF HIGH CELL DENSITY, PERFUSION PROCESSES FOR CONTINUOUS MANUFACTURING.

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Presentation on theme: "| www.genzyme.com Jade (with her mother) Fabry disease USA UPSTREAM DEVELOPMENT OF HIGH CELL DENSITY, PERFUSION PROCESSES FOR CONTINUOUS MANUFACTURING."— Presentation transcript:

1 | Jade (with her mother) Fabry disease USA UPSTREAM DEVELOPMENT OF HIGH CELL DENSITY, PERFUSION PROCESSES FOR CONTINUOUS MANUFACTURING Tim Johnson, Ph.D. October 21, 2013

2 Perspectives on Continuous Manufacturing Upstream Development −Steady-State Control −Approach to Process Development −Scale-Up Conclusions Discussion Points

3 Continuous Integrated Biomanufacturing Drivers Predictable Performance Simplicity UniversalStandardization Flexible Core Drivers Manufacturing, Process, & Business Drivers Reduced Tech Transfer Risks Efficient time Steady State Processes & Product Quality Reduced Footprint Variable Steady state Quality indicator Variable Problem

4 Capture Intermediate Purification Polish Clarified Harvest Bioreactor Media Harvest Hold Clarification Unform DS Perfusion Fed-Batch Current State – Biomanufacturing Processes Limited Standardization, large and complex

5 Capture Clarified Harvest Bioreactor Media Harvest Hold Clarification Perfusion Continuous Biomanufacturing Action Steady-State High Cell Density High Productivity Key Technology High Sp. Production Rate Low Perfusion Rate

6 Continuous Biomanufacturing Action Benefit Steady-State High Cell Density High Productivity Capture Clarified Harvest Bioreactor Media Harvest Hold Clarification Perfusion Reduced Bioreactor Size SUBs now feasible Standardized Size Universal – mAbs/Enz Key Technology High Sp. Production Rate Low Perfusion Rate

7 Continuous Biomanufacturing Action Benefit Continuous flow Bioreactor  Capture Capture Bioreactor Media Perfusion Removes: Hold steps Clarification Ops. Simplified Process Key Technology Simultaneous Cell Separation and Clarification

8 Continuous Biomanufacturing Action Benefit Continuous capture Capture Bioreactor Media Perfusion Reduced column size and buffer usage Key Technology Periodic Counter-Current Chromatography

9 Capture Bioreactor Media Future State – Continuous Biomanufacturing Standard, Universal, Flexible Integrated Continuous Biomanufacturing Unform. Drug Substance Unform. Drug Substance Predictable Performance UniversalStandardization Flexible Reduced Tech Transfer Risks Efficient time Steady State Processes & Product Quality Reduced Footprint Variable Steady state Quality indicator Variable

10 Future State – Continuous Biomanufacturing Standard, Nearly Universal, Flexible PAT & Control Process Knowledge Robust Equipment & Design Facilitating Aspects Predictable Performance UniversalStandardization Flexible Reduced Tech Transfer Risks Efficient time Steady State Processes & Product Quality Reduced Footprint Variable Steady state Quality indicator Variable

11 Steady-state cell density Steady-state nutrient availability  Steady-state metabolism  Steady-state product quality Steady-State Upstream Control VCD Cell Specific Perfusion Rate = Perfusion Rate Cell Density Viable Cell Mass Indicator

12 Cell Density Control Strategies 12 r 2 = 0.88r 2 = 0.73 r 2 = 0.70 Viable Cell Mass Indicators  Capacitance  Oxygen sparge  Oxygen uptake rate  Others

13 Steady cell density and growth Steady-State Upstream Demonstration Steady-state metabolism Steady-state production and product quality CQA #3 Volumetric Productivity CQA #1 CQA #2

14 Glycosylation Profiling Steady-State Product Quality Over 60 days Peak 1Peak 4Peak 5 Peak 7Peak 8Peak 11

15 break- even OPEX drivers for continuous biomanufacturing Vs. fed-batch −High cell density −High volumetric productivity High Cell Density – High Productivity mAb Demonstration −Low perfusion rate −Low media cost Viable cell density Cell-Specific Perfusion Rate OPEX Savings Favorable to Perfusion VCD Productivity Volumetric Productivity (g/L-d)

16 Perspectives on Continuous Manufacturing Upstream Development −Steady-State Control −Approach to Process Development −Scale-Up Conclusions Outline PAT & Control Process Knowledge Robust Equipment & Design

17 F1 F2 F3 F4 SET 1SET 2SET 3SET 4 40 weeks Unrealistic timelines required to study full process (60 days/run) Leverage steady-state to condense experiments Process Development Design of Experiments S.S. Perfusion Fed-batch ~11-15 weeks 15 weeks SET 1SET 2SET 3SET 4 F1 F2 F3 F4 Measure response shift SET 1SET 2SET 3SET 4 F1 F2 F3 F4

18 Approach −Four factors determined from screening studies − Cell Specific Perfusion Rate − pH − Dissolved Oxygen − ATF Exchange Rate −Custom design with interaction effects  24 conditions Process Development Design of Experiments ATF Exchange Rate

19 Design of Experiments Results Culture generally stable over the ranges tested Cell Specific Perfusion Rate is the most significant factor Little interaction effects SPR Growth Rate Viability Product Quality #1 Cell Specific Perfusion Rate pH DO ATF Exchange Rate

20 Operational Space Determine acceptable operational space −Fixed cell specific perfusion rate ATF Exchange Rate Acceptable Space pH Out of Spec Regions Green – Viability Red – Growth rate Blue – Product Quality #1 Dissolved Oxygen

21 Reactor Productivity Capture Yield Combined Productivity Optimum pH Integrated Operating Spaces Example  Integrating upstream and downstream process knowledge  Upstream: Productivity ↓ below critical pH value  Downstream: Yield recovery ↓ as pH ↑ Solution  Optimal pH exists to maximize productivity and yield Productivity Yield pH

22 Perspectives on Continuous Manufacturing Upstream Development −Steady-State Control −Approach to Process Development −Scale-Up Conclusions Outline PAT & Control Process Knowledge Robust Equipment & Design

23 Scale-up to Single Use Bioreactor Skid −Custom HyClone 50L Turnkey System −Bioreactor customized for perfusion −Nine control loops Scale-up approach −Match scale independent parameters −Accounted for scale dependent parameters − Agitation: match bulk P/V Initial Run −Conservative 40 Mcells/ml set-point −60+ day operation −10L satellite running concurrently SUB ATF

24 Scale-up Results Growth and Metabolism Cell Density Oxidative Glucose Metabolism Growth rate and metabolism are as expected

25 Scale-up Results Productivity ProductivityProduct Quality #1 Productivity and product quality are as expected

26 Scale-up Results Continuous Chromatography Integration Capture operation using three column PCC −Fully automated −Steady-state performance UV ChromatogramSDS PAGE for Capture Elution Harvest Day DS Warikoo, Veena, et al. Integrated continuous production of recombinant therapeutic proteins. Biotech. & Bioeng. v109, ; 2012 Godawat, Rahul, et al. Periodic counter-current chromatography – design and operational considerations for integrated and continuous purification of proteins. Biotech. Journal v7, ; 2012 S.S. Harvest Feed Consistent Capture Duration and Frequency

27 Reactor Scale Considerations Productivity Possibilities 50L can meet some low demand products 500L can meet average demand products * Kelly, Brian. Industrialization of mAb production technology: The bioprocessing industry at a crossroads. mAbs 1:5, ; 2009 * 50L 500L Further optimization #

28 Summary and Conclusions  Core drivers achieved  Achieved robust and steady-state control  Developed methodology for efficient process understanding  Successfully scaled-up upstream process to 50L SUB  Platform routinely being applied to mAbs and Enzymes  Simplicity and design for manufacturability considerations are a cornerstone of our continuous & integrated platform  Additional challenges remain Simplicity

29 Genzyme/Sanofi Industrial Affairs Late Stage Process Development Commercial Cell Culture Development Purification Development Process Analytics Early Process Development Analytical Development Translational Research Many other colleagues at Genzyme GE Healthcare Acknowledgements


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