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Vellore 2011 Vellore 2011 Presentation Transcript

  • “Current Best Practise in Biomanufacturing and thePlatzhalter Bild Critical Role of Innovation” International Vellore Symposium “Bioprocess Industry-Academia Interaction” July 2011 Dr. Uwe Gottschalk, VP Purification Technologies, Sartorius Stedim Biotech
  • What are the hot Topics? 7th Annual Survey of Biopharmaceutical Manufacturing. Eric S. Langer, BioPlan Associates Inc.
  • Existing Facilities to Meet Current Challenges View slide
  • The USDP/DSP Interface in a World of High Titers Data adapted from: F. Wurm Production of recombinant Protein Therapeutics in Cultivated Mammalian Cells. Nature Biotechnology 22, 1-6 (2004) View slide
  • DSP is Mass not Volume driven Jim Davis, Lonza Economics of Monoclonal Antibody Production: The relationship between upstream titer and downstream costs; IBC San Diego March 2008
  • Current Best Practise in DSP High Titer Implications: Increasing biomass and contaminant levels Protein A pool volumes and step cost DNA & HCP levels post Capturing Polishing load volumes and conductivity Pathogen clearance as a moving target
  • Chromatography Technologies for DSP Polishing (Membranes) • Highly porous structure • Pore size: 3 – 5μm • Convective Flow Capturing/IP (Resins) • Minimal buffer use • Bead size distribution: 15 -160 μm • Average pore size: 15 - 40 nm • Diffusion limited flow • High capacity
  • Capture Costs: Why bother? Jim Davis, Lonza Economics of Monoclonal Antibody Production: The relationship between upstream titer and downstream costs; IBC San Diego March 2008
  • Emerging capture technologies expected to have limited market potential inupcoming years Technology Description Maturity Risk Relevance • CIM, BIA Separations, methacrylate based monoliths Monoliths • Similar to membrane adsorbers • Purification of large biomolecules (viruses, plasmid DNA, conjugates), good resolution Expanded • Upfront/DSM work on single use technology for MABs Capture and intermediate purification bed ad- • Already used in depletion of valuable biomolecules from particle • No significant sorption containing feedstreams at large scale (milk, juice, etc.) market in upcoming years expected – • Membrane adsorber technology Potential for Direct Capture • Own development and IP, depletion of valuable biomolecules from Niches (e.g. MA particle containing feedstreams Vaccines, DNA) • Currently low priority, combines cell harvest and capture chromatography step Ligands • instAction, Prometic (mimetic ligands), BAC, GEHC pipeline, Repligen Protein A Precipitation/ • Used at large scale in plasma fractionation (precipitation) Crystallization/ and APIs (crystallization) • No immediate Extractionn • Not developed for mABs commercialization possible • Potential for Affinity • Polybatics disruptive nano- • Disruptive technology technology particles • Single use alternative to Protein A • Platform character Already used In development Start of development Low risk Moderate risk High riskSource: Sartorius
  • Alternatives to Protein A Capture• Product precipitation batch/continuous• Impurity precipitation (followed by non-Protein A process)• Alternative Capturing (Protein A Mimetics, Mixed Mode, CEX)Issues: Selectivity, Scale up, Reproducibility, Comparability
  • addresses: Protein A pool volumes and step costD. Low BioManufacturing Paris 2007
  • Alternative Protein A Chromatography Formats:Goal: Intensified Use/Volume Reduction• Simulated Moving Bed (SMB) and related: » Tarpon („single use flow path“) » Novasep » Chromacon » Chromatan » ... ______________________________________________________________________• Expanded Bed Chromatography » DSM/Upfront („single use flow path“)Issues: Complexity, Scale up, Reproducibility, Comparability
  • Alternative Protein A Formats:Goal: Low Cost – Real Single Use 2000: Oleosin Platform 2005: TMV Nanoparticles 2010: Bio Polyester Platform Polyester Synthase Polyester Granule Immunoabsorbent nanoparticles based on a 100-300 nm tobacco mosaic virus displaying protein ALimitation: Oleosin yields < 1kg/ha S. Werner et al. PNAS 103, 17678 - 17683 Grage, K. and Rehm, B.H.A. (2008) Bioconj. Chemistry, 19(1):254-62.
  • Two Birds – one Stone: Contaminant Precipitation at Pfizer and Medarex addresses: Fig 7a. precipitation based process Fig 7b. TFF based process Contaminant precipitation Protein A pool volumes TFF and step cost DNA & HCP levels post HCP < 1000 ng/mg CEX HCP < 10ng/mg CEX Dilution Capturing Dilution Q Membrane Q Membrane HCP < 1000 ng/mg HCP BDL 2 g/ml 20 g/ml Dilution VF Mix Mode VF Process Scale Precipitation of Precipitation of Process-Derived Impurities in Mammalian Cell Culture Impurities in Non-Protein A Broth; J. Glynn et al. In: Gottschalk U Purification Schemes for MAb; J. Wang (ed) Process-scale Purification of et al. BioPharm Intl. 10/2009, 2-9 Antibodies. Wiley, NY.
  • Chromatography Technologies for DSP Polishing (Membranes) • Highly porous structure • Pore size: 3 – 5μm • Convective Flow Capturing/IP (Resins) • Minimal buffer use • Bead size distribution: 15 -160 μm • Average pore size: 15 - 40 nm • Diffusion limited flow • High capacity
  • Pete Gagnon 2007
  • Selection Guide Convective Media Q,S Low salt Polishing in flowthrough: STIC viruses, DNA, Host ng is hi cell proteins, Pol High salt endotoxins, HIC aggregates Convective Media Cap Purification: large t ure proteins (Factor Q,S VIII), viruses (vaccines), phages...
  • Sartobind STIC® - Next Generation of Membrane AdsorbersShares same cellulose base membrane as Q: >3 μm pore size0.5 μm Here is the binding capacity Sartobind Q Grafted Quaternary ammonium Sartobind STIC Direct derivatisation Binding capacity + ligand density is distributed more evenly + pore accessibility Primary amine (Sartobind STIC PA) I. Tatárova, I., R. Fáber, R. Denoyel, M. Polakovic, J. Chromatography A 1216 (2009) 941March 7, 2011 Page 9
  • Host cell protein removal from up 10 kg mAb per L (pH 8, 500 ppm HCP load, 10MV/minApplication Note Sartorius-Stedim Biotech: 85032-540-18, 05/2011
  • Sartobind STIC Sartobind Q Sartobind STICBinding Capacity (g/m²)BSA 2 mS/cm @ 0 mM NaCl 9 19BSA 20 mS/cm @ 200 mM NaCl 1 10DNA 7 mS/cm @ 50 mM NaCl 2 6Removal of ΦX174 (LRV)LRV 1.4 mS/cm @ 0 mM NaCl 3,7 5,1LRV 6.7 mS/cm @ 50 mM NaCl 0,1 5,1LRV 16.8 mS/cm @ 150 mM NaCl 0,1 4,8Removal of MVM (LRV) at Wuxi ApptecTrial 1 16.8 mS/cm @ 150 mM NaCl 2.10 3,82Trial 2 16.8 mS/cm @ 150 mM NaCl 1.81 >4,96
  • Source: 2nd Annual Survey of the Bioprocessing Marketfor Single-Use SolutionsAspen Brook Consulting, 2010
  • Current Challenge in UF • Process efficiency should be HIGH • Membrane cleaning should be EASY • Final Mab concentration may reach 20% • Maximum system pressure is LIMITED E ECO Pumping Requirements 8 L/m2/min 2 L/m2/min Viscosity High Low MAb Concentration >15% <10%
  • Select the Right Products Flux vs. Concentration for Mab Sartorius ECO & E cassette Crossflow Rate: 360 L/m2-Hr 100 90 ECO 80 eate FLux (LMH) 70 60 50 40 E Perm 30 20 10 0 10 100 1000 Concentration (g/l)
  • Yes we can!
  • Upcoming Alternatives Przybycien, Pujar, Steele: Current Opinion in Biotechnology 2004, 15 469-478
  • Disruptive Technologies from Inception to Maturation Konstantinov, K. Towards fully continuous bioprocessing: What can we learn from Pharma? Cell Culture Engineering XII, Banff, Canada (2010)
  • Trend in New Drug Production Scales
  • Flexible and Disposable
  • New Facilities to meet Future Challenges
  • Thank you!Uwe.Gottschalk@sartorius-stedim.com