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A new, integrated, continuous purification process template for monoclonal antibodies

A new, integrated, continuous purification process template for monoclonal antibodies. Alex Xenopoulos* Alison Dupont, Christopher Gillespie , Ajish Potty, Michael Phillips Processing Technologies Merck Millipore Bedford, MA (USA). Integrated Continuous Biomanufacturing A new ECI conference

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A new, integrated, continuous purification process template for monoclonal antibodies

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  1. A new, integrated, continuous purification process template for monoclonal antibodies Alex Xenopoulos* Alison Dupont, Christopher Gillespie, Ajish Potty, Michael Phillips Processing Technologies Merck Millipore Bedford, MA (USA) Integrated Continuous BiomanufacturingA new ECI conference Castelldefels, Spain October 20-24, 2013

  2. Highlights • We developed a flow-through purification train that enables an integrated, continuous process • We have novel solutions for continuous clarification and capture • Bench-scale proof of principle for several mAbs shown • Breakthrough improvements not possible unless you look at new technologies AX | ECI Castelldefels | 21Oct2013

  3. Monoclonal antibody production • A mature, robust industry • Templated process • Protein A chromatography • Yet, several issues remain • Stability • Capital and utilities • Large footprint • Frequent bottlenecks • Sterility • Cleaning validation AX | ECI Castelldefels | 21Oct2013

  4. New alternative template 2° depth filtration Protein A b/e chrom CEX b/e chrom AEX f/t chrom Virus filtration Current template UF/DF Clarification Capture Purification/polishing Bioreactor Centrifuge Protein A b/e chrom continuous Carbon f/t device AEX f/t device CEX f/t device 1° depth filtration Bioreactor w/ precipitation Alternative template AX | ECI Castelldefels | 21Oct2013

  5. Comparison of templates – icons sized by device volume Current template Clarification Capture Purification/polishing 3.3 m2 14.1 L 14.1 L 19.3 L 0.6 L each 4.4 m2 5 L 0.4 L 3 L Alternative template 1,000 L @ 2 g/L AX | ECI Castelldefels | 21Oct2013

  6. Comparison of templates – pool tanks Current template 500 L 1000 L Clarification Capture Purification/polishing 250 L Alternative template 50 L AX | ECI Castelldefels | 21Oct2013

  7. Status Three launched Clarisolve™ filters optimized for particle size Portfolio of flocculants Continuous harvesting and loading of protein A column successful and beneficial Benefits Elimination of centrifuge up to 6,000 L Increased throughput (<3x membrane area) DNA removal (1-2 LRV) Advantages persist post protein A Reduced turbidity Enhanced HCP clearance Reduced resin cleaning Clarification assisted by precipitation and using novel Clarisolve™ filters results in post-Protein A benefits AX | ECI Castelldefels | 21Oct2013

  8. Capture with continuous multicolumn chromatography and incompressible Protein A resins offers savings • Status • Two incompressible resins available • Prosep® Ultra Plus • Eshmuno® A • Continuous loading from clarified harvest and continuous loading to purification train successfully shown • Benefits • Higher productivity, especially at low residence times • Resin and buffer savings AX | ECI Castelldefels | 21Oct2013

  9. Protein A capture cannot be beaten as part of a holistic process evaluation • Why not CEX chromatography? • Cheaper resin • Cheaper unit operation • Two dilution steps – volume increase • Longer processing time • Higher water/buffer use • Lower selectivity • Less virus removal • Lower yield • Increased process development • Less templatable • More expensive • Why not precipitation? • Single-use • Buffer consumption • Processing time • More materials • Additional unit operations • Precipitant removal • No product concentration • Dilution steps • No purification • Increased process development • More expensive at commercial scale AX | ECI Castelldefels | 21Oct2013

  10. Purification in flow-through mode using novel adsorbers, minimum interventions, fewer pool tanks and one skid VF withprefiltration CEX b/e AEX f/t Traditional Process AEX Pool CEX Pool VF Pool Low pH VI Pool Carbon +AEX f/t CEX f/t +VF Proposed Process Low pH VI Pool VF Pool In-line pH AX | ECI Castelldefels | 21Oct2013

  11. Novel flow-through adsorber functionalities work synergistically to remove several classes of impurities mAb Aggregates CEX Larger acidic HCP, DNA, viruses AEX • Cell culture components • Insulin, methotrexate, Pluronic F68®, hygromycin, antifoam C • Process-related impurities • DNA, HCP, leached Protein A, viruses • Product-related impurities • Aggregates, fragments MAb Low MW high Low MW impurities (leached Protein A, HCP, fragments) Carbon acidic pI basic AX | ECI Castelldefels | 21Oct2013

  12. Benefits of flow-through purification • Disposable chromatography devices connected without pool tanks • No bind/elute chromatographic steps • Minimal interventions • Orthogonal mechanisms for impurity removal • Needed pH adjustments incorporated in skid • One skid (protein A elution  TFF) is possible • Enables integrated, continuous process template AX | ECI Castelldefels | 21Oct2013

  13. Internal bench-scale experimental case studies: Robustness of flow-through purification train (3 mAbs) AX | ECI Castelldefels | 21Oct2013

  14. External trials:Robustness of flow-through purification train (7 mAbs) AX | ECI Castelldefels | 21Oct2013

  15. Internal case studies:Product quality AX | ECI Castelldefels | 21Oct2013

  16. Cost of Goods: where is the advantage? AX | ECI Castelldefels | 21Oct2013

  17. Process modeling: advantages of proposed template AX | ECI Castelldefels | 21Oct2013

  18. Key features of the alternative template • An alternative templated process for downstream purification of mAbs is proposed • It matches performance of current templates, provides operational advantages • Features: • Novel downstream purification process for mAbs – from bioreactor through formulation • Connected unit operations – continuous operation, minimal interventions • Novel unit operations developed – leverage continuous nature • Clarification toolbox – novel depth filters, precipitating agents • Product capture with continuous multicolumn protein A affinity chromatography – efficient use of resin and buffer • Flow-through polishing – no bind/elute steps, improved simplicity and economics • Virus filtration and ultrafiltration/diafiltration – no changes • Proof of concept and feasibility data generated – performance equivalent to current, advantages in overall operational flexibility AX | ECI Castelldefels | 21Oct2013

  19. Acknowledgments • Downstream Technologies, MM • Kevin Galipeau • Meghan Higson • Jad Jaber • Mikhail Kozlov • Matthew Stone • William Cataldo • Romas Skudas • Jeff Caron • Jonathan Steen • Scott Bliss • Dennis Aquino • Wilson Moya • Analytical Technologies, MM • Rong-Rong Zhu • Michael Bruce • Team Supply, MM • Michael McGlothlen • Patricia Kumpey • Paul Hatch Business Development, MM • Fred Mann BioPharm Services, Inc • Andrew Brown AX | ECI Castelldefels | 21Oct2013

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