UPSTREAM DEVELOPMENT OF HIGH CELL DENSITY, PERFUSION PROCESSES FOR CONTINUOUS MANUFACTURING

Transcription

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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 Product Quality #1 Growth Rate Viability Cell Specific Perfusion Rate ph DO ATF Exchange Rate

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

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

22 Outline Perspectives on Continuous Manufacturing Upstream Development Steady-State Control Approach to Process Development Scale-Up Process Knowledge Conclusions PAT & Control 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 SUB Initial Run ATF Conservative 40 Mcells/ml set-point 60+ day operation 10L satellite running concurrently

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

25 Scale-up Results Productivity Productivity Product 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 Chromatogram SDS PAGE for Capture Elution DS Harvest Day S.S. Harvest Feed Consistent Capture Duration and Frequency 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

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

28 Summary and Conclusions Simplicity 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

29 Acknowledgements 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