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Platform downstream processes in the age of continuous chromatography: A case study

Platform downstream processes in the age of continuous chromatography: A case study. Mark Brower BioProcess Technology & Expression Bioprocess Development Kenilworth, NJ. Integrated Continuous Biomanufacturing Castelldefels , Spain 20-24 October 2013. Transition to Future Concepts.

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Platform downstream processes in the age of continuous chromatography: A case study

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  1. Platform downstream processes in the age of continuous chromatography: A case study Mark Brower BioProcess Technology & Expression Bioprocess Development Kenilworth, NJ Integrated Continuous Biomanufacturing Castelldefels, Spain 20-24 October 2013

  2. Transition to Future Concepts To meet increasing global demands requires… PROCESS INTENSIFICATION Batch Stainless Batch Stainless / Single Use Continuous Single Use Enabled Next Generation

  3. Primary Recovery (Centrifugation / MF + DF) Bulk Purification Protein A Chromatography Viral Inactivation (Low pH Hold) DNA / HCP / Viral Adsorption Anion Exchange Chromatography Variant and Aggregate Clearance Cation Exchange Chromatography Viral Filtration Nanofiltration Concentration / Buffer Exchange Microfiltration / Diafiltration Bioburden Reduction Sterile Filtration mAb Downstream Purification 6H Bulk 12H 18H 24H • Increased flexibility • Reduced footprint • Reduced capital spend • Better resource utilization 30H Fine 36H 42H 48H Formulation 54H 60H

  4. p p p p p p Continuous Processing Vision - 2,000L SUB* Formulation: BRF/DiaF Continuous UF A E X M Anion Exchange Membrane Surge Bag S U B* BRF p Viral Filtration Continuous Viral Inactivation p Single-Use Centrifugation Surge Bag Surge Bag Surge Bag Surge Bag Depth /BRF Filtration Polishing Step BioSMB Protein A BRF BRF Overall DSP Time Cycle is Dictated by the Longest Step Other Steps are Lengthened to Compensate *Single-Use Bioreactor

  5. Continuous Processing Case Study mAb 1- Non-platform AEX Membrane SMB Protein A SU Centrifuge Harvest Bag SPTFF SUB DF / BRF Viral Inactivation Mixed Mode

  6. C1 C2 C4 C3 Strip Elute Eq Wash 2 Product Waste Depleted Feed Wash 2nd pass 2nd pass Feed C6 C8 C5 C7 MCC for Bind & Elute Applications • Methods based on batch process • Loading, washing, elution, CIP carried out simultaneously • Flexibility in loading zone Switch Time

  7. CEX CMCC Load Zone Design Feed 2nd Pass Feed Longer residence time in the elution zone Similar column cycling compared with protein A Productivity 3.7X batch process 2nd Pass W1 W1 • 2 methods designed to maximize time in the elution zone • Wash 1 in parallel 8 columns (shorter / continuous feed) • Wash 1 in series 6 columns (longer / discontinuous feed)

  8. SMB Transformation of Platform CEX Step • 1.2cm x 3cm pre-packed columns • Poros HS Adsorbent • qbatch=50mg/mL • Feed = 11-13g/L • 2 different load zone configurations • Good agreement between experimental and theoretical capture efficiency • CMCC loading was 60-73mg/mL at high yield >95% 3.7 x Specific Productivity Design Equations* *Miyauchi and Vermeulen (1963)

  9. Aggregate Clearance – Wash in Series Configuration • Effect of column height investigated • 1.2 x 3.4cm, 1.2 x 6.8cm, 0.5 x 20cm • Feed aggregation varied (low and high) • Six 1.2 x 3.4cm columns for MCC • 4th cycle fractionation (20 fractions per column pooled) • Similar pre-peak observed in batch and MCC Process • Similar pool aggregate levels observed • Little difference observed at different column heights

  10. Integration of MCC CEX into Continuous DSP- 100L platform harvest p p p p p p Formulation: BRF/DiaF CRITICALITY pH Continuous UF Continuous UF A E X M Surge Bag S U B* BRF p Viral Filtration VI p Surge Bag Surge Bag Surge Bag Surge Bag Depth /BRF Filtration BioSMB CEX BioSMB Protein A BRF BRF

  11. Continuous CEX Performance Column • 16 Overlaid CEX Elution Profiles • AEXM Effluent Feed STDEV(%) Between Columns =1.01%

  12. Continuous Processing Case Study mAb 2- Platform Mass Balance = 93%

  13. DSP Productivity Enhancement • MCC steps enjoy a modest specific productivity increase • Other steps suffer from lower specific productivities because they are slowed to accommodate the incoming flow rate • The overall DSP will be 2-4x more productive (g/day) by operating in parallel (dependent on Protein A column sizing)

  14. Conclusions and Future Work • A platform cation exchange step was transformed into a MCC process • 3.6X specific productivity increase • Maintained consistent aggregate separation performance compared to the batch process • Integrated into continuous DSP top reflect platform operation with 84% yield at the 100L scale • Matched cycles with protein A step • Interface CEX step with continuous viral filtration • Scale up process to 2000L in 24hours

  15. Acknowledgements • BTE • Ying Hou • David Pollard • Analytical Support • Joe Fantuzzo • John Troisi • Jun Heo • Fermentation Support • Patty Rose • Chris Kistler • Rachel Bareither • Protein Purification Process Development • Nihal Tugcu • Thomas Linden • Marc Bisschops • Steve Allen

  16. Questions?

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