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Platforms for mAb Commercialization
The document discusses advancements in mammalian antibody (mAb) commercialization, highlighting KBI Biopharma's capabilities in cell line development, manufacturing, and analytical services. It emphasizes the shift from clinical to commercial process development and the integration of high-throughput systems for efficient product development cycles. Additionally, it outlines the importance of robust scientific understanding and innovative chromatography techniques in optimizing mAb processes for scalability and productivity.
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Platforms for mAb Commercialization
- 1. Platforms for mAb commercialization AbhinavA. Shukla, Ph.D. Senior Vice President Development & Manufacturing KBI Biopharma, Durham NC BPI West, San Francisco, February 27 – March 2, 2017
- 2. Durham, North Carolina -Cell Line Development - Cell Culture cGMP Manufacturing - Analytical QC, Formulation, Stability - Mass Spec Core Facility RTP, North Carolina - Mammalian Process Development - Analytical development Boulder, Colorado - Cell Line Development - Microbial Process Development - Microbial cGMP Manufacturing - Analytical, QC, Formulation, Stability - Particle Characterization Core Facility The Woodlands, Texas - Cell Therapy Manufacturing - Cell based assays Contract Development & Manufacturing Organization
- 3. Programs in mammalianclinical manufacturing at KBI • Primary emphasis 2011 – 2016 on clinical entry stage programs • ~ 10-14 IND filings per year supported via development & manufacturing efforts • Also supported several stand-alone programs for process characterization studies • Now emphasis shifting to include commercial launch process development & manufacturing for mammalian cell culture programs • Boulder Colorado site already commercial ready for microbial products
- 4. KBI Biopharma Upstream Train I UpstreamTrain II ProA VI Polish VF Bulk fill KBI’s Cell Culture Manufacturing Facility Purification Suite 2000L Prodn BRX200L Seed BRXWaveSF Harvest 2000L Prodn BRX200L Seed BRXWaveSF Harvest
- 5. 3/9/20175 | Mammalian cell culture expansion • Leveragingsignificant capabilities in cell line development, analytical methods • Development, formulation development & process development • Adding additional 2000L bioreactor in train II • Dedicated downstream purification for each train • Expanded buffer and media preparation integral with commercial suite • Increasing capacity to > 50 batches per year 0 5 10 15 20 25 30 2013‐14 2015 2016 New Products Mfg Batches Cell Culture Manufacturing in KBI Biopharma (Durham, NC)
- 6. 6 Biologics Commercialization Pre-Clinical PhaseI Phase II Phase III Process Development Process Characterization Process Validation Process Monitoring & Improvement FIH Process • Deliver clinical process quickly • Platform process • Clinical Supply Submission & Approval Lifecycle management BLA Prep & PAI Commercial Process • Deliver manufacturing process for registrational trials and market • Design keeping large-scale manufacturing in mind • Improve productivity, efficiency, robustness, manufacturability, COGs • Analytical characterization and method development Process Characterization and Validation • Develop IPC strategy through understanding of process inputs and outputs (design space) • Scale-down characterization and validation studies • Large-scale process validation to demonstrate process consistency • BLA preparation • Supporting documents for licensure inspections • Post-commercial process improvements (CI) • Post-commercial process monitoring FIH process Commercial process
- 7. 7 Improvements in platformtechnology can enable one process development cycle • Essential for biosimilars or highly accelerated programs • Streamlined process characterization/validation effort Process Characterization Lifecycle Management Platform Application IND BLA Commercial Platform Technology Development
- 8. 8 Outline • Where aremAb platforms today and do they enable a single cycle of development? • FIH process • Commercial process • Can a platform approach be taken for commercialization • Process characterization studies leading to definition of an in- process control strategy (IPC) – how fast can these be completed? • Scale-down validation studies • Conformance lots • Commercialization in single-use manufacturing facilities
- 9. 9 Investments in fundamental understandingof bioprocesses • Robust platforms can only be developed if there is a strong understanding of the science of developing bioprocesses • Multimodal chromatography • Platforms for non-mAbs (HIV vaccine proteins) • Improved Protein A resins • Creates the ability to react quickly if an “unusual” observation is made • All process decisions need to be made keeping large- scale production in mind
- 10. 10 0 50 100 150 200 250 300 350 400 450 500 0.0% 20.0% 40.0%60.0% 80.0% 100.0% HCP (ppm) Recovery Capto MMC HCP Clearance 25mM Tris pH 7.0 (baseline) 25mM Tris pH 7.0, 5% ethylene glycol 25mM Tris pH 7.0, 50mM arginine 25mM Tris pH 7.0, 50mM NaSCN 25mM Tris pH 7.0, 1M urea 25mM Tris pH 7.0, 1M ammonium sulfate 25mM Tris pH 7.0, 0.1M NaCl 25mM Tris pH 7.0, 0.5M ammonium sulfate 25mM Tris pH 7.0, 0.1M NaCl, 1M urea Washes that disrupt protein-protein interactions Conventional washes log k’ = A – Blog(csalt) + C(csalt) k’ = (tr – tm )/tm Wolfe, L., Barringer, C., Mostafa, S., Shukla, A. Multimodal chromatography: characterization of protein binding and selectivity enhancement through mobile phase modulators, Journal of Chromatography A, 1340, 151-156, 2014. Multimodal chromatography
- 11. 11 High capacity ProteinA chromatography resins Resin Vendor Matrix Ligand Modified Protein A domain Mean particle size (µm) MabSelect SuReTM GE Healthcare Highly cross‐linked agarose Alkali‐stabilized rProtein A B domain 85 MabSelect SuReTM LX GE Healthcare Highly cross‐linked agarose Alkali‐stabilized rProtein A B domain 85 ToyopearlTM AF‐rProtein A HC‐650F† Tosoh Polymethacrylate Alkali‐stabilized rProtein A C domain 45 EshmunoTM A† EMD Millipore Cross‐linked Polyvinyl Ether Alkali‐stabilized rProtein A C domain 50 AmsphereTM A3† JSR Life Sciences Polymethacrylate Alkali‐stabilized rProtein A C domain 50 1 2 3 4 5 6 0 10 20 30 40 50 60 70 80 mAb2 DBC(g/L) Residence Time (min) MabSelect SuRe MabSelect SuRe LX rProtein A HC-650F Eshmuno A Amsphere A3 mAb1 mAb2 mAb3 mAb4 0 1000 2000 3000 8000 9000 10000 HCPLevel(ppm) MabSelec SuRe MabSelect SuRe LX rProtein A HC-650F Eshmuno A Amsphere A3
- 12. CH505 Envelopes selectedas vaccine immunogens CH505 transmitted-founder (TF) and Env variants generated during viral evolution drove affinity maturation of CH103 bnAb lineage Antibody: UCA T/F gp120 Kd = ~200 nM Env: CH103 CH505 wk53 CH505 wk78 CH505 wk100 CH103 lineage intermediate antibodies CH505 TF CH505 wk136 12 H.X. Liao et al. Nature 496: 469; 2013
- 13. • Parameters shaded in gray are defined across molecules. Parameters shaded in yellow require molecule specific optimization • For all Envmolecules the operating parameters, basal medium, feed type and some of the supplement additions have defined • The need for additional supplements is molecule specific • Reasons for supplement addition: » Biocompatability in SU bioreactors » Increase in productivity Scale Temperature Set point 37.0 ± 0.5°C Temperature Shift 33.0 ± 0.5°C on Day 6 DO Set point 30% pH Set point 6.90 ± 0.1 Agitation (1-impeller) 50 rpm → 55 rpm Air overlay 1.6 SLPM Air Sparge 0.5 SLPM Max. Oxygen Sparge 5 SLPM Max. CO2 Sparge 5 SLPM Medium CD OptiCHO + 8 mM Glutamine Target VCD 0.50 x 106 cells/mL Base 1M Sodium carbonate Feed Type: LTI Feed A+B (1:1) 15% on Day 0, 10% current wv each on Days 3, 6, and 9 Supplement 1 addition: HT Supplement 1X current wv each on Days 0 and 4 Supplement 2 addition: Cystine Supplement 3 addition: Tyrosine Supplement 4 addition: Soy:Yeastolate Hydrolysate (2:3) 5g/L current wv each on Days 4 and 8 Supplement 5 addition: C1615 Harvest Add 10g/L Hydrolysate on harvest
- 14. • Parameters shaded in gray are defined across molecules. Parameters shaded in yellow require molecule specific optimization • Load and elution conditions for three of the unit operations require molecule specific definition given the heterogeneity of this class of molecules •Env antigens structurally sensitive to hydrophobic surfaces, hence HIC not employed
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- 17. 17 Next generation mAbplatforms • Driver • High cell culture productivity is increasing interest in ultra- high loading polishing steps (> 100 mg/mL loading) Genentech Biogen Millipore proposal Protein A Viral Inactivation Cation-exchange chromatography Anion-exchange chromatography Viral Filtration UF/DF Protein A Viral Inactivation AEX flowthrough No salt Hydrophobic Interaction Chromatography flowthrough Viral Filtration UF/DF Protein A Viral Inactivation Anion-exchange flowthrough Overloaded cation- exchange chromatography Viral Filtration UF/DF
- 18. 18 Which platform shouldI use?
- 19. 19 Next generation mAbplatforms • Platform processes for mAbs have hugely facilitated the growth of mAbs as therapeutic agents • Rapid clinical entry with lower cost & resource burden & significant time savings (gene to IND in ~ 12-14 months) DS Process Platform DS Process Platform Cell line diversity Cell line diversity Media/feed type diversity Media/feed type diversity HCP level variability HCP level variability Cell density variability Cell density variability HMW level variability HMW level variability Protein A Viral Inactivation AEX Based Polishing (Flow Through mode) CEX Based Polishing Viral Filtration UF/DF
- 20. 20 Multimodal chromatography innext generation mAb platforms • Mixed-mode has the simultaneous ability to clear HMW and HCP leading to mAb platforms with wider coverage • Added advantage of ability to operate over wider conductivity range for loading 93.0 94.0 95.0 96.0 97.0 98.0 99.0 100.0 0.0 10.0 20.0 30.0 40.0 50.0 60.0 70.0 80.0 90.0 100.0 Capto S ImpAct pH 5.0 Eshmuno CPX pH 5.0 Fractogel SO3 pH 5.0 Capto MMC pH 7.5 Selectivity Curves for HMW Clearance MainPeak(%) Increase selectivity Accumulated Yield (%) Hydrophobicity scale: Capto S < Fractogel SO3 < POROS HS50 < Nuvia cPrime < Capto MMC
- 21. 21 Success of mAbplatforms that include multimodal chromatography • Can successfully accommodate wide range of cell lines and cell culture feed streams into a single downstream platform • Cell lines from KBI, Bioceros, Selexis, Cellca, Excellgene, Antitope, Life Technologies 0.0% 1.0% 2.0% 3.0% 4.0% 5.0% 6.0% 7.0% 8.0% ProA AEX CEX BDS %HMW mAb Platform HMW Clearance mAb A mAb B mAb C mAb D mAb E mAb F mAb G mAb H mAb I 0 5000 10000 15000 20000 25000 ProA AEX CEX BDS rHCP(ppm) mAb Platform rHCPClearance mAb A mAb B mAb C mAb D mAb E mAb F mAb G mAb H mAb I
- 22. 22 MAPPING PROCESS DESIGN SPACE(PROCESS CHARACTERIZATION AND SCALE-DOWN VALIDATION)
- 23. 23 Quality by Design(QbD) • “Quality by design means designing and developing manufacturing processes during the product development stage to consistently ensure a predefined quality at the end of the manufacturing process.” ICH Q10, FDA 2006 Process Design (Process Development) Process Control Strategy Definition Process Validation Continued Process Verification
- 24. 24 Process Design Space Higher level of assurance of product quality Manufacturing Efficiency and Flexibility Continuous process improvement while maintaining product quality Characterization Space Design space Control space Design Space (ICH Q8, 2006): The multidimensional combination and interaction of input variables (e.g., material attributes) and process parameters that have been demonstrated to provide assurance of quality. Need a high throughput scale-down model for the process
- 25. 25 Accelerating the EntireProduct Development Lifecycle
- 26. 26 Accelerating process characterization& scale-down validation studies • Small-scale bioreactors (1-10L working volume) have been the traditional scale-down model in industry till date • Accelerating PC/PV studies requires a high- throughput scale-down model • Ambr250 as a scale-down model for cell culture processes
- 27. 27 Mimicking “Cellular Environment”in SDMs. Given the large sets of variables in a cell culture process, establishing a cost and time-efficient SDM, mimicking a cellular environment similar to large scale production bioreactor, is critical for conducting successful PC studies.
- 28. 28 Matching key processindicators in SDMs Comparison of time courses for viable cell growth and lactate profiles for two recombinant CHO cell lines in ambr™ SDMs for a mAb and a Biosimilar. Matching cell growth and lactate profiles for CHO cell lines producing a mAb and Biosimilar respectively were key process indicators and in turn dictated the process yield and product quality.
- 29. 29 Comparison of SDMsacross scales
- 30. 30 Accelerated Upstream PCTimelines with high-throughput SDMs Month 1.5 SDMQ USP Month 5.5 N-1/N-2 Screening (40 x 3L Seed) Harvest PC Work 12 -15 Harvest conditions Month 0 Raw Materials and Worst Case (20 x 3L and 1 round of ambr250 runs: 24 vessels) Main Stage PC (3 rounds of ambr250 runs: 72 vessels) Inoculum Studies (100 Shake Flasks and 4 Wave Runs) Worst-case Linkage USP/DSP Month 7.0
- 31. 31 Scale-Down Process ValidationStudies • Scale-down validation studies in addition to large- scale process validation (conformance lots) • Probe extremes in the process and demonstrate them to be acceptable • Examples • Reprocessing validation – combine hold times with process conditions that create the greatest stress on the protein • Intermediate hold times – combine hold times and demonstrate releasable drug substance • Viral clearance studies • Impurity clearance studies
- 32. 32 Conclusions • Robust scientificunderstanding is a pre-requisite for developing robust platforms that can make single cycle development possible • Highly generic and manufacturing friendly mAb platforms have been designed (gene to IND in 12-14 months) • Process characterization & scale-down validation studies can be accelerated (6-9 months) by using high throughput cell culture platforms as the scale-down model • Single-use manufacturing is now a viable commercial launch platform (long term may combine single use manufacturing & higher productivity continuous manufacturing)
- 33. 33 mAb platforms SU manufacturing Highthrough process development tools Rapid process characterization & validation tools & approaches Henry David Thoreau 1817-1862, writer Walden Pond Mahatma Gandhi 1869 - 1948
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