High Productivity Membrane Chromatography: Enabling the Next Generation Bioprocessing Paradigm
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A novel single-use chromatography platform enabling cost-efficient manufacturing to support the growing global demand for affordable biologics. A new single-use (per batch) chromatography platform employs traditional, proven chemistries in an inventive hydrogel polymer membrane format that enables the next generation bioprocessing paradigm. The experimental Protein A membrane, featuring a 10-fold improvement in productivity over resin columns and high selectivity (i.e. 3 LRV HCP), is combined with high performance membranes with mixed mode and ion exchange modalities for a fully single-use membrane-based purification process. The membrane columns show protein capacities similar or superior to reference resins in bind and elute, and up to 7 LRV clearance of MVM at 20kg/L in flow through mode. A membrane-based process allows a holistic process strategy involving small footprint, high throughput processing by means of a rapid multi-cycling capture step (up to 100 cycles per batch) and high capacity flow through polishing. This single-use (per batch) platform results in simple, compact, flexible, yet robust and safe downstream operations for cost-efficient manufacturing to support the growing demand for affordable biologics. In this webinar, you will learn about: • Advantages of a fully single-use membrane-based purification processes • High capacity flow through polishing with an inventive membrane adsorber combining the best of resins (DBC) and membranes (30x of flow rate)
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High Productivity Membrane Chromatography: Enabling the Next Generation Bioprocessing Paradigm
- 1. Merck KGaA Darmstadt, Germany Renaud Jacquemart, Ph.D. Principal Scientist Director Vaccines Process Sciences Enabling the next generation bioprocessing paradigm High productivity membrane chromatography
- 2. 2 The life science business of Merck KGaA, Darmstadt, Germany operates as MilliporeSigma in the U.S. and Canada.
- 3. Next Generation Bioprocessing Paradigm
- 4. Next Generation Processing Any technology, expendable, service, or system which significantly changes the existing monoclonal antibody manufacturing template to deliver: Speed Flexibility Quality Cost4
- 5. Performance Drivers Flexibility Reduce product change- over time by 90% Speed Reduce facility construction, mAb production and product release times Cost 90% reduction in cost to manufacture and capital expenditure Quality Increased robustness and reliability BioPhorum Operations Group (BPOG) Technology Roadmap The Industry Requires Step Change in Performance Process Analytics Process Intensification Single Use Enablers 5
- 6. Driver Metric Current State 5-yr target 10-yr target Flexibility Facility utilization <70% >85% >95% Product changeover time 3 days <18 hours <8 hours Time to reconfigure suite for new process >2 weeks <1 week <2 days Speed Facility build speed 3 years 2 years 1 year Time to make product 4-6 months 2 months 1 month Quality Cost of quality >10% of operating cost 10% of operating cost 2% of operating cost Inventory cover 3-6 months 2 months 2 weeks Cost Total cost to supply $100/g $50/g $10/g Cost of facility construction $500M+ $100M $50M From the 2017 BioPhorum Operations Group (BPOG) Technology Roadmap (Page 18, Table 1) Next Generation Processing Tough Goals: Reduce Facility Cost and mAb CoGs New technologies & processing paradigms required6
- 7. Facilities are the biggest Cost Driver… but they depend on the Process! Holistic Approach to Biomanufacturing 7 Total cost of ownership NPC (including facility construction costs, capital, and operating costs with depreciation) for mAb production at 2000 L scale with fed-batch process (3 g/L titer) ▪ Facility cost was 40% of cost of ownership ▪ Implementation of single-use technologies reduced Net Product Cost by 40% Pralong & Pollard (2017) Single-Use Technology Implementation For Biologics and Vaccines Production, p.724 Cost drivers in traditional facilities 7
- 8. Incentives, Challenges and Pathways 8 Low CoGs needed to The challenge? Supply developing countries with affordable biologics Reach emerging markets profitably Improve competitiveness and long-term sustainability Enable business model for new biologics and novel therapies (political pressure on pricing, cost is a real factor) Great progress in USP (high titer, perfusion reactors) DSP still done the traditional way: slow, low yield, large expensive units USP & DSP not integrated Facilities are large, expensive, not flexible and long lead time to construct Holistic Approach to Biomanufacturing Better Processes = Smaller Facilities = Lower Costs
- 9. Consensus in the Industry? Next Generation Processing is enabled by High Productivity 9
- 10. A High Productivity DSP that matches Intensified USP 10 What is our Proposal? TRADITIONAL APPROACH • Resin columns need oversizing to match USP productivity Large footprint, not flexible, labor/quality-intensive • High CAPEX & OPEX Needs multi-batch amortization NEW PARADIGM • High Productivity DSP • Increased flexibility at lower CapEx • Cost efficient solution to match high USP productivity
- 11. What is our Proposal? High Productivity Membrane Chromatography 11 High Productivity Membrane Chromatography • High throughput achieved without over-sizing • Utilization of rapid multi- cycling to reduce media volume • No amortization needed • Single use per batch bind & elute Protein A • Right-sized flow-through salt-tolerant CEX with HIC modality • High loading flow-through salt-tolerant AEX Bioreactor CEX AEX Primary separation Capture Intermediate / polishing VF UF/DF Protein AFiltration
- 12. Poll 1
- 14. High Productivity Chromatography Membrane High Binding & Short Residence Time Natrix® HD membrane 1. Reinforcing mesh “skeleton” • Provides mechanical strength & durability to composite membrane • Polypropylene – good chemical stability 2. Porous polymer gel • 3D macroporous structure provides: • A large surface area that contains a high density of protein binding groups • Interconnected pores that provide convective flow channels 14
- 15. Natrix® Membranes Flow and Binding Dynamics (1/2) Combining the best of Resin & Membrane Natrix® Membranes o Single use o Dominated by advective flow o High binding capacity for proteins, virus and DNA o High flow rates Conventional Column Chromatography Natrix® Advective Chromatography Resins o High protein binding capacity o Limited capacity for large targets (virus, DNA) o Diffusional limitations o Low flow rates Conventional Membranes o Single use o High flow rates o Limited surface area o Low binding capacity Conventional Membrane Adsorber Chromatography 15
- 16. Natrix® Membranes Flow and Binding Dynamics (2/2) Binding Capacity maintained over wide Residence Time range (1.2s - 12s) Membrane: maximum 10% loading capacity loss for 10x faster loading Resin: loading capacity decreased by 80% when residence time decreased by 4x (86s to 21s) 0 20 40 60 80 100 120 0 5 10 15 NormalizedLoadingcapacity% Residence Time (seconds) Membrane Resin Natrix® membranes are flow rate insensitive Operate at 6s residence time or less with no capacity loss 16
- 17. Natrix® Affinity Membranes Composition & Properties NatriPur™ HD-A: high productivity Protein A [NOT YET COMMERCIAL] • Developed for rapid multi-cycling Bind & Elute operation • Productivity (g mAb/h.L) 30-fold higher than Protein A resins • Demonstrated for 20+ mAbs • Single-use per batch • Caustic stable • Reactive membrane platform can accommodate multiple types of ligands • Demonstrated for virus and recombinant proteins purification 17
- 18. Natrix® Ion Exchange Membranes Composition & Properties NatriPur™ HD-Sb: salt-tolerant CEX with HIC modality [NOT YET COMMERCIAL] • CEX group – Sulfonic acid • HIC group – t-Butyl • High binding capacity and salt tolerance Flexible operating conditions • Caustic stable (1M NaOH) • Autoclavable 18
- 19. Natrix® Ion Exchange Membranes Composition & Properties NatriFlo® HD-Q: salt-tolerant AEX [COMMERCIALLY AVAILABLE] • AEX group – Quaternary amine • High binding capacity • Buffers and salt tolerance Flexible operating conditions • Caustic stable (1M NaOH +2M NaCl) • Autoclavable (Gamma-stable product in development) 19
- 20. Poll 2
- 21. The Natrix® Paradigm Shift
- 22. New template enabled by Natrix® membrane technologies Traditional mAb process template Natrix® Membranes Towards The New Paradigm 22
- 23. Towards The New Paradigm Natrix® Membranes Unlock A New Way Of Processing: Case Studies Enabling fully single-use process and true flexible facilities Rapid multi-cycling, right-sizing and full utilization of media life in 1 batch Designed for flow-through operation to further increase productivity Modular and Flexible: can be scaled out rather than scaled up 1 2 3 4 23
- 24. Towards The New Paradigm Natrix® Membranes Unlock A New Way Of Processing: Case Studies Enabling fully single-use process and true flexible facilities Rapid multi-cycling, right-sizing and full utilization of media life in 1 batch Designed for Flow-Through operation to further increase productivity Modular and Flexible: can be scaled out rather than scaled up 1 2 3 4 24
- 25. 25 Rethink Protein A Options exist to replace Protein A… BUT constraints on process development time and the need for a robust manufacturing are strong incentives to keep but rethink Protein A Enabling Fully Single-Use Processes (1/3)
- 26. HCP clearance for Protein A membrane comparable to reference resins 26 Enabling Fully Single-Use Processes (2/3)
- 27. 27 • NatriPur™ HD-Sb & NatriFlo® HD-Q provide state-of-the-art chromatography performance and are single use • All other unit operations are already single use • HD-A Capture is the missing link • Once implemented: all benefits of flexibility will be unlocked Enabling Fully Single-Use Processes (3/3) True fully flexible facilities become reality… BUT how to handle chromatography cost? Bioreactor HD-Sb HD-Q Primary separation Capture Intermediate / polishing VF UF/DF HD-AFiltration
- 28. Towards The New Paradigm Natrix® Membranes Unlock A New Way Of Processing: Case Studies Enabling fully single-use process and true fully flexible facilities Rapid multi-cycling, right-sizing and full utilization of media life in 1 batch Designed for Flow-Through operation to further increase productivity Modular and Flexible: can be scaled out rather than scaled up 1 2 3 4 28
- 29. Cycling Study mAb 8: 50 cycles @ 20 g/L Load Avg Leached ProA <2ppm Avg Delta Column Pressure <2psi Media life fully utilized within a batch improves economics 29 High Productivity Enables Rapid Multi-Cycling & Right-Sizing (1/4)
- 30. High Productivity Enables Rapid Multi-Cycling & Right-Sizing (2/4) 30 Up to 5kg mAb produced over 10 days Perfusion bioreactor • 100 L • 2 volumes/day • Titer: 2.5 g/L Biosimilar for developing country • Cost must be low CV: 23.9L BC: 35 g/L CV: 1.2L BC: 35 g/L MV: 0.5L BC: 45 g/L High flow rate to match perfusion productivity BUT too expensive with long and uncertain amortization Expend full life of media/batch cost efficient BUT too slow to match perfusion rate Meets perfusion rate with no need to oversize Rapid Multi-Cycling • Expend full life of media/batch Cost Efficient Traditional, large column approach Residence time (sec): 240 Cycle time (min): 180 Hours per day: 3h every 2 days Cycles over 10 days: 5 Cost-efficient resin column Residence time (sec): 240 Cycle time (min): 180 Hours per day: 28.4 Cycles over 10 days: 110 Right-sized membrane column Residence time (sec): 6 Cycle time (min): 12 Hours per day: 4.5 Cycles over 10 days: 200
- 31. 31 Daily purification train matching perfusion flow rate MV: 0.5L Load 45g/L 20 cycles 2.5 hours 95% yield 90% yield 98% yield HD-Sb Polish HD-Q Polish HD-A Capture MV: 0.5L Load 300g/L 3 cycles 1.5 hour MV: 0.03L Load:12500gg/L 1 cycle 3.5 hours Adapted from Jacquemart et al., CSBJ, 2016 4 suites in a 2,000 m2 pod-based micro-facility costing <€20M 500 kg mAb purified annually! High Productivity Enables Rapid Multi-Cycling & Right-Sizing (3/4) CoGs <$50/g compatible with biosimilar markets in developing countries enabled by right-sizing and rapid multi-cycling Right-sized high productivity Protein A membrane capture (400 g/h•L-media) (30X resin productivity) Straight-Through Processing in subsequent high throughput DSP steps 4.5 kg purified for every batch (10 days) in 20 ft2 of GMP suite.
- 32. Benoit Mothes (Sanofi) Fully automated, high productivity DSP Multi-unit operation system with 3 types of Natrix® membranes • Consistent performance (100 cycles) with & without sanitization every cycle • Sanitization not required since columns are disposable? Source: “ASAP: Toward a Fully Disposable Continuous Process”, Mothes et al., BPI Conference 2014 32 Adrian Gospodarek (MSD) First demonstrations of continuous multi- membrane chromatography SMB multi-column system with 5 Natrix® test cells • 91 cycles • Load: 30 g/L @ 4 sec RT • Avg yield: 87% • Avg HCP reduction: 4.1 LRV • DNA in eluate: <1 ppm Source: “High Capacity Protein A Membranes for mAb Capture: An Alternative to Column Chromatography”, Gospodarek et al., 2016 AIChE Annual Meeting Continuous purification platforms further optimize productivity High Productivity Enables Rapid Multi-Cycling & Right-Sizing (4/4)
- 33. Towards The New Paradigm Natrix® Membranes Unlock A New Way Of Processing: Case Studies Enabling fully single-use process and true fully flexible facilities Rapid multi-cycling, right-sizing and full utilization of media life in 1 batch Designed for Flow-Through operation to further increase productivity Modular and Flexible: can be scaled out rather than scaled up 1 2 3 4 33
- 34. 34 Designed for Flow-Through Operation To further Increase Productivity (1/3) HCP = 247 ppm Aggregates = 10.35% Yield = 93% HCP = 47 ppm Aggregates = 0.49% Yield = 93% HCP = 3 ppm Aggregates = 0.42% NatriPur™ HD-Sb pH 5.5 10 mS/cm Load: 300 g/L NatriFlo® HD-Q pH 7.5 2 mS/cm NatriPur™ HD-Sb: optimized hold-up volume to enable multi-cycling mode (even in FT operations) NatriFlo® HD-Q: designed for single cycle, extended load at high flow rate and low pressure drop 34
- 35. 35 Designed for Flow-Through Operation To further Increase Productivity (2/3) HCP = 1123 ppm Aggregates = 1.91% Yield = 88% HCP = 162 ppm Aggregates = 0.75% Yield = 96% HCP = 26 ppm Aggregates = 0.74% NatriPur™ HD-Sb pH 7.5 4 mS/cm Load: 300 g/L NatriFlo® HD-Q pH 7.5 4 mS/cm 2 flow through steps provide further optimization of media utilization and DSP time Improved productivity and process economics No feed adjustment between the flow-through steps for simple, continuous operation 35
- 36. Feed: ProA & CEX Purified mAb • Titer: 10g/L • HCP: 70 ppm Condition: 25mM Tris, pH 7.5, 10ms/cm NatriFlo® HD-Q FT pool achieves • HCP <10ppm • No detectable DNA • 7 LRV MVM • Great purification performance at very high loading capacity = Downsize the unit • Performance maintained at high conductivity = Reduce operations & risks 36 Designed for Flow-Through Operation To further Increase Productivity (3/3)
- 37. Towards The New Paradigm Natrix® Membranes Unlock A New Way Of Processing: Case Studies Enabling fully single-use process and true fully flexible facilities Rapid multi-cycling, right-sizing and full utilization of media life in 1 batch Designed for Flow-Through operation to further increase productivity Modular and Flexible: can be scaled out rather than scaled up 1 2 3 4 37
- 38. • Combination of multi-cycling and modular units : only the number of cycles and/or number of columns change • No technical or regulatory scale up required • Minimizes investment when risks are high (early phases) • Maximizes PD efficiency when moving through late stages • Scaling out enables seamless transition from early stage to late stage / manufacturing Modularity Enabled By Productivity38 SCALING OUTSCALING UP Modular and Flexible Can be Scaled Out rather than Scaled Up (1/2)
- 39. 39 800 32 Phase 1 Batch size (g) # of cycles Batch processing time (hrs) Phase 2 Phase 3 Commercial No change in unit operation: no process development or regulatory work From bench to bedside: one size fits all! 2000 80 4000 80 8000 80 80 80 80 80 Modular and Flexible Can be Scaled Out rather than Scaled Up (2/2) 2 4.9 5 5.1
- 40. Poll 3
- 41. Conclusions
- 42. Cost savings are confirmed even for large scale manufacturing 42 Reproduced from Pollard et al., BPI, 2016 The BioSolve® model developed by MSD supports earlier conclusions
- 43. Natrix® High Productivity Membrane Chromatography Enabling the Next Generation Bioprocessing Paradigm 1 Critical quality attributes comparable to reference resin processes 2 Single use, high productivity chromatography enables • Fully single-use, modular processes • Unlocking true flexible facilities that promote better facility utilization • Rapid multi-cycling, right-sizing, full utilization of media life in 1 batch 3 Cost-efficiency is achievable for both clinical and commercial manufacturing by realizing a holistic strategy 43
- 44. New paradigm enabled by Natrix® membrane technologies Traditional mab process template The future
- 45. The vibrant M, Natrix, NatriPur, and NatriFlo are trademarks of Merck KGaA, Darmstadt, Germany or its affiliates. All other trademarks are the property of their respective owners. Detailed information on trademarks is available via publicly accessible resources. © 2018 Merck KGaA, Darmstadt, Germany and/or its affiliates. All Rights Reserved. Renaud Jacquemart, Ph.D. Director, Vaccines Process Sciences Natrix Separations, Inc. 5295 John Lucas Drive – Unit 6 Burlington, ON Canada L7L6A8 renaud@natrixseparations.com Cell: 289-828-0728