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Complete single-use ADC technology from development through scale-up
The document discusses the growth of the antibody drug conjugates (ADC) market, projecting a CAGR of 21.82% from 2017-2022 due to increasing cancer cases and a robust pipeline of ADC drugs. It outlines the advantages of single-use technology in ADC production, including safety, efficiency, and cost-effectiveness, and details various Mobius® systems for ADC processing, such as mixing and chromatography. Additionally, it emphasizes successful scaling up of bioprocess chemistry for ADC-X, aiming for consistency and high yield for clinical trials.
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Complete single-use ADC technology from development through scale-up
- 1. Merck KGaA Darmstadt, Germany GangYao, Ph.D. Principal Scientist Mary Robinette Principal Project Engineer Kevin Reyer Project Engineer
- 2. 2 The life sciencebusiness of Merck KGaA, Darmstadt, Germany operates as MilliporeSigma in the U.S. and Canada.
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- 4. The Future ofADCs 4 According to the research report, "Global Antibody Drug Conjugates (ADC) Market - Analysis By Drugs (Adcetris, Kadcyla), Pipeline Drugs, Regulations: Opportunities and Forecasts (2017-2022)", global market is projected to display a robust growth represented by a Compound annual growth rate (CAGR) of 21.82% during 2017-2022, chiefly driven by large number of ADC drugs in pipeline, rising number of cancer patients and wider therapeutic window offered by antibody drug conjugates. (https://www.researchandmarkets.com/research/q6jlz9/glo bal_antibody)
- 5. ADC Process Flow Single-UseTechnology can be applied to all ADC processing steps mAb mAb Rxn Mix Final BDS Crude ADC Purified ADC mAb Intermediate Dilution / Buffer Exchange (UF, Filter) Reaction #1 – mAb Modification (Reactor) Reaction #2 – Payload Conjugation(Reactor) Purified ADC UF/DF (UF, Filter) Chromatography (Chromatography, Filter) Formulation (Filter) UF/DF + Formulation (UF, Filter) 5
- 6. 6 Single-Use Technology: TheBenefits Safe • Minimizes operator exposure to potent materials Integral • Decreased risk of contamination • Validation, quality and regulatory compliance support Novel • Process automation enables reproducible commercial scale quantities • Highly customized solutions from our expansive component library Greater Flexibility • Closed, sterile sampling system enables representative samples • Compact footprint and mobile carrier construction for enhanced flexibility Lower Cost • Single-use flow paths provides maximum adaptability to your changing operational needs • Eliminates cleaning validation Efficient • Easy setup and use • Reduced down-time for cleaning • Broad working volumes
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- 8. Mobius® Single-Use JacketedMixing System ADC Single-Use Reactor sizes Custom Jacketed Mobius® Mixer design Sizes: 10 L, 50 L, 100 L, 200 L, 500 L, 1000 L Solvent compatible materials Sample ports Instrumentation Temperature pH Batch weight Mixing speed Mobius® Single-Use Jacketed Mix 100 L 8
- 9. Turnover Time ScaleUp Volume (L) 0.8 8 80 Reactor 1 L Glass 10 L single-use 100 L single-use Mixer Speed (rpm) Turnover Time (min) Turnover Time (min) Turnover Time (min) 75 0.25 0.45 1.16 135 0.14 0.25 0.65 350 0.05 0.10 0.25 Mobius® Single-Use Jacketed Mixing System: Mixing Scale Up Geometrically similar scale up Typical mixing parameters Power per unit volume Impeller pumping rate/Turnover time 53 DnNpPower 9
- 10. Mobius® Single-Use JacketedMixing System: Mixing Studies Blend Time Setup Blend time Volume 10%, 50%, 100 Mixer speed 25%, 50%, 100% Shear Particle formation % aggregate 10
- 11. Mobius® FlexReady Chromatography:ADC Purification Pump cart capability Low flow: 0.1 – 2.2 L/min High flow: 1.6 to 8 L/min Dual pumps: accurate gradient Pressure up to 4 bar 2 x 5 inlets and 4 outlets 11 Mobius® FlexReady Solution with Smart FlexwareTM Assemblies
- 12. Mobius® FlexReady Chromatography:ADC Purification Batch Size Flexibility Range of pump flowrate 3 flowpath sizes, optimized for flow rate Accurate Gradient Capability <2% error over flow range, 10-90% Flow control pump speed basis conductivity basis Gradient at 0.5 L/min 12
- 13. Mobius® FlexReady Chromatography:ADC Purification Operator Interface Accurate Column Qualification Minimal axial dispersion of injected solution Achieve tight asymmetries and low HETP Full automation and data collection 21 CFR Part 11 compliant 13
- 14. Mobius® FlexReady TFF:ADC Purification Mobius® FlexReady TFS2 skid Cross flow: 2 – 18 L/min Membrane area up to 5m² Retentate 50L and 100L mix bag Pressure up to 4 bar Mobius® FlexReady TFS3 skid Cross flow: 4 – 40 L/min Membrane area up to 10m² Retentate 200L mix bag Pressure up to 4 bar 14 Mobius® FlexReady Solution with Smart FlexwareTM Assemblies
- 15. Mobius® FlexReady TFF:ADC Ultrafiltration Batch Size Flexibility Range of crossflow and membrane area Retentate tank volume 10-100% for active mixing 0-10% mixing via pump recirculation Retentate return through diverter plate Eliminates retentate bypass to feed Low concentrate volume Dye Test - Retentate Diverter Plate 15
- 16. Mobius® FlexReady TFF:ADC Ultrafiltration High Product Recovery Low system holdup volume Sloped piping with zero dead-leg valves 4 step recovery process with air or buffer Full automation and data collection 21 CFR Part 11 compliant Product Recovery Optimized Flush 16
- 17. 17 Applicability of single-useTechnology for ADCs Compatibility/Extractable Studies including ADC solvents single-use Assembly Testing Leak testing LAL and particulate Class VI USP Product Specific Testing BioReliance® Extractable and Leachable Validation Services Solvent Component
- 18. After 24 hourexposure to 20% DMSO and 20% DMAc No leaks and no physical changes observed by visual inspection Smart FlexwareTM and connectors passed all functionality testing Extractables from 20% DMSO and 20% DMAC samples at ppb levels Solvent Testing – ADC single-use Assemblies Recovery Line To Transfer Pump Smart Flexware® Assembly for TFF 100L Single-use Feed Container Assembly Filtrate Assembly with Single-use Sensors Flow Cell Retentate Sampling Port Assembly Dow Corning® Pharma-80 Tubing, used to create bypass loop around TFF liners and cassettes Extractable Test Setup on Mobius® FlexReady Solution with Smart Flexware® Assemblies for TFF Single-use Feed Pump Assembly 18 Extractable Test Setup on Mobius® FlexReady Solution with Smart FlexwareTM Assemblies for TFF TM
- 19. 19 Case Study • Technologytransfer and optimization of bioprocess chemistry and associated analytical methods to enable the production of antibody-drug conjugate bulk drug substance ADC-X with 1000 gram antibody under cGMP. • This material will be used as an Active Pharmaceutical Ingredient for human Phase 1 clinical trials. • The ADC-X construct features a novel IgG1 isotype antibody conjugated to a new drug-linker.
- 20. 20 Development Strategy • Theprimary goal of the development work was to deliver consistent and scalable bioprocess chemistry that routinely affords functional ADC-X with the desired final product specifications in high yield. • This objective was achieved by performing evidence driven development campaign resulting in the production of a pilot / toxicology batch, followed by the technology transfer to cGMP production.
- 21. Conjugation Reactors Demonstration Batch (1L Glass Reactor) Toxicology Batch (10 L SU Reactor) GMP Batch (100 L SU Reactor) 21
- 22. Temperature in ConjugationReactors SolutionTemp(°C) Time (hour) 10 L for Toxicology Batch 100 L for GMP Batch 22
- 23. Scale Up Basedon Turnover Time Can be Achieved for 1 L Glass Reactor, 10 L single-use Reactor and 100 L single-use Reactor Turnover time verses Mixer Speed and Reactor volume Working Volume (L) 0.8 8 80 Reactor 1L Glass 10L Singe Use 100L Single Use Mixer Speed rpm 1 L Glass Reactor (min) Mix 10 turnover (min) Mix 100 turnover (min) 25 0.74 1.34 3.49 40 0.46 0.84 2.18 55 0.34 0.61 1.59 75 0.25 0.45 1.16 100 0.18 0.34 0.87 135 0.14 0.25 0.65 150 0.12 0.22 0.58 170 0.11 0.20 0.51 200 0.09 0.17 0.44 260 0.07 0.13 0.34 350 0.05 0.10 0.25 500 0.04 0.07 0.17 Mixing in Conjugation Reactors 23
- 24. Similar HPLC 1Chromatograms Between Crude ADC from Demonstration Batch and Toxicology Batch Crude ADC from Demonstration Batch and Toxicology Batch – HPLC 1 24
- 25. Crude ADC fromDemonstration Batch and Toxicology Batch – HPLC 2 Similar HPLC 2 Chromatograms Between Crude ADC from Demonstration Batch and Toxicology Batch 25
- 26. Preparation Chromatograms FromDemonstration Batch and Toxicology Batch Demonstration Batch Toxicology Batch Similar Separation Profiles Obtained between Demonstration Batch and Toxicology Batch 26
- 27. Similar HPLC 1Chromatograms Between Final ADC from Demonstration Batch and Toxicology Batch Comparison of Final ADC from Demonstration Batch and Toxicology Batch – HPLC 1 27
- 28. Comparison of FinalADC from Demonstration Batch and Toxicology Batch – HPLC 2 Similar HPLC 2 Chromatograms Between Final ADC from Demonstration Batch and Toxicology Batch 28
- 29. Analytical Results ofFinal ADC from Demonstration Batch and Toxicology Batch Test Demo Batch Tox Batch Appearance - Color B6 not more intensely colored than B6 Appearance - Clarity 9.5 10.4 Appearance - Particles comparable pH within 0.1 unit difference Protein within 1.0 mg/mL difference DAR within 0.1 unit difference Monomer comparable Aggregate comparable Unconjugated mAb comparable Finger Print 1 comparable Finger Print 2 comparable Finger Print 3 comparable Osmolality comparable Residual Free Drug Linker comparable Endotoxin 0.011 EU/mg 0.007 EU/mg Bioburden (TYMC/TAMC) TYMC 0 CFU/20 mL TAMC 0 CFU/20 mL TYMC 0 CFU/20 mL TAMC 0 CFU/20 mL Comparable Analytical Results Obtained for Products from Demonstration Batch and Toxicology Batch All Test Results Met Target Specifications 29
- 30. 30 Process Evaluation • ConjugationProcess Chemistry scaled up from Demonstration Batch to Toxicology Batch. o Excellent agreement of the mass balance and product quality attributes o Alignment of the product quality attributes with the target values o Cumulative data on process scalability and consistency • Provided a decision on proceeding to Technology Transfer to GMP Manufacturing.
- 31. Preparation Chromatograms FromToxicology Batch and GMP Batch Toxicology Batch GMP Batch Similar Separation Profiles Obtained between Toxicology Batch and GMP Batch 31
- 32. Comparison of FinalADC by HPLC 1 Similar HPLC 1 Chromatograms Between Final ADC from Different Batches 32
- 33. Comparison of FinalADC by HPLC 2 Similar HPLC 2 Chromatograms Between Final ADC from Different Batches 33
- 34. Test Demo BatchTox Batch GMP Batch Appearance - Color B6 not more intensly colored than B6 not more intensly colored than B6 Appearance - Clarity 9.5 10.4 10.1 Appearance - Particles comparable pH within 0.1 unit difference Protein within 1.0 mg/mL difference DAR within 0.1 unit difference Monomer comparable Aggregate comparable Unconjugated mAb comparable Finger Print 1 comparable Finger Print 2 comparable Finger Print 3 comparable Osmolality comparable Residual Free Drug Linker comparable Endotoxin 0.011 EU/mg 0.007 EU/mg 0.006 EU/mg Bioburden (TYMC/TAMC) TYMC 0 CFU/20 mL TAMC 0 CFU/20 mL TYMC 0 CFU/20 mL TAMC 0 CFU/20 mL TYMC 0 CFU/20 mL TAMC 0 CFU/20 mL Analytical Results of Final ADC Comparable Analytical Results Obtained for Products from Demonstration Batch, Toxicology Batch and GMP Batch All Test Results Met Target Specifications Process Successfully Scaled up from Demonstration Batch to Toxicology Batch, then Transferred to GMP Production 34
- 35. Mobius® Technical support •R&D • Applications Engineering • Bioprocessing Bio-Conjugation Center of Excellence St. Louis • Global Process & Analytical Development • ADC Production manufacturing
- 36. Summary We offer abroad range of single-use equipment, custom designed with our experts to meet the challenging needs of ADC production. ADC processes can be proportionally scaled up with single-use equipment. ADC made with single-use equipment met the target quality attributes. 36
- 37. The Vibrant Mis a trademark 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.