Scalability of a Single-Use Bioreactor Platform for Biopharmaceutical Manufacturing
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Presented at PepTalk 2017: San Diego, CA Niket Bubna, Principal Scientist, Process Development, KBI Biopharma Single-use Technologies And Continuous Processing (Advancing Bioprocessing Through Technological Innovation) Risk Mitigation Strategies For Single-use Technologies
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Scalability of a Single-Use Bioreactor Platform for Biopharmaceutical Manufacturing
- 1. Niket Bubna Principal Scientist, Process Development, KBI Biopharma Durham, NC Presented at PepTalk 2017: San Diego, CA Single-use Technologies And Continuous Processing (Advancing Bioprocessing Through Technological Innovation) Risk Mitigation Strategies For Single-use Technologies
- 2. Jan 2017PepTalk 2017 0 5 10 15 20 25 30 2013-14 2015 2016 New Products Mfg Batches Cell Culture Manufacturing in KBI Biopharma (Durham, NC) Reference: Medicines in Development (Biologics). 2013 PhRMA Report
- 3. • New biopharmaceuticals being developed at a record rate • Rapid development and manufacturing for FIH studies is needed to support the rate of discovery • Tox and FIH studies require smaller quantity of product • Commercial needs can be met with 2000 L-scale bioreactors leveraging high titer cell culture processes • Need for a readily available plan and plant • Plan includes process, equipment and supply chain • Equipment and supply chain are available • Scalable SUB platform needed Jan 2017PepTalk 2017
- 4. Reference: Shukla, A., Mostafa, S., Wilson, M., Lange, D. Vertical Integration of Disposables in Biopharmaceutical Drug Substance Manufacturing, Bioprocess International, 10(6), 34-47, 2012.
- 5. Jan 2017PepTalk 2017 Typical Cell Culture Manufacturing Unit Operations at KBI (Durham, NC)
- 6. • Single-use bioreactors are being used for clinical manufacturing and even commercial manufacturing • Clinical manufacturing » Wave for ~2 decades » Stirred tank 2000 L-scale since 2009 • Commercial manufacturing » Amgen, Singapore • Multiple SUB platforms are available • HyClone HyPerforma • Sartorius BIOSTAT STR • Millipore Mobius Jan 2017PepTalk 2017
- 7. • Minimal cleaning requirements • Quick turnaround leading to higher plant capacity • Flexibility of manufacturing multiple products • Fewer requirements for changeover • Reduced risk of cross-contamination Jan 2017PepTalk 2017
- 8. Jan 2017PepTalk 2017 • Agitation Rate • Homogenous mixing is important to provide uniform environment around the cell • Power input per volume is commonly used to normalize agitation rate based on the power input to the impeller • Gassing Strategy • Variables: Sparge type, sparge hole size and gas flow rate • Use of VVM (vessel volumes per minute) or the ratio of volumetric gas flow to the liquid volume is used; it depends on the actual culture volume, allowing operations at multiple volumes
- 9. • Combination of three key factors: • Power input per volume • Volumetric gas flow rate • Bioreactor geometry Jan 2017PepTalk 2017 𝑃𝑃 𝑉𝑉 = 𝑁𝑁𝑝𝑝 𝑁𝑁3 𝐷𝐷𝑖𝑖 5 𝜌𝜌 𝑉𝑉 where, Di = Impeller diameter [m] N = Agitation speed [s-1] Np = Impeller power number [-] P = Power [W] V = Volume [L] 𝑉𝑉𝑉𝑉𝑉𝑉 = 𝑄𝑄𝐺𝐺 𝑉𝑉 where, QG = gas flow rate [m3/s] V = Volume [L] VVM = Vessel volumes per minute [min-1] Parameter XDR-50 SUB XDR-200 SUB XDR-2000 SUB Impeller Diameter 0.2159 m 0.2159 m 0.4191 m Impeller Power Number 1.50 1.15 0.72 Pitched-blade Impeller 3 blades at 40° 3 blades at 40° 4 blades at 40° Turn-down Ratio 2.2:1 5:1 5:1 Aspect Ratio 1.5:1 1.5:1 1.5:1
- 11. Jan 2017PepTalk 2017 Project Cell Line Type Cell Line Vendor Basal Medium Doubling Time (hours) Forward Processing VCC (10^6 cells/mL) Passage Duration Can Platform Approach Be Used? A CHO-DG44 A & KBI OptiCHO / Dynamis 25-35 2-3 3-4 days No B CHO-K1 B & KBI ProCHO 5 21-24 1.7-2.5 3 days No C-F CHO-S A & KBI Dynamis 14-20 3-7.5 3-4 days Yes G Proprietary C BalanCD Growth A 18-21 3.5 3 days Yes H CHO-DG44 D BalanCD Growth A 16-17 4 3 days Yes I CHO-GS E FortiCHO 25-40 None 2-4 days No J CHO-GS F CD CHO 20-24 3 3 days No K Proprietary C BalanCD Growth A 15-20 2-6 3 days Yes L CHO-GS G Client Medium 37-42 3 3-4 days No M CHO-GS F OptiCHO / FortiCHO 22-23 3 3 days Yes N Proprietary (CHO-K1) H OptiCHO / PowerCHO-GS 20-30 3.5 4 days No O Proprietary C BalanCD Growth A 17-21 3.5 3 days Yes P CHO-DG44 E ExCell CHO 30-40 1.5 2-3 days No Q-S CHO-DG44 A & KBI OptiCHO 20-30 2.5-4.5 3-4 days Yes T CHO-DG44 D ExCell ACF CHO 21-24 4 3-4 days No U Proprietary I OptiCHO 25-35 1.5-5 3-4 days Yes
- 14. Jan 2017PepTalk 2017 Rapid process development to support IND filing & FIH Studies • Top Candidate Clone to final cell culture process in 3.5 months • Applied platform process (medium-feed and process parameters) • Required feed media change to support increase in titer • Process scale-up and cGMP manufacturing completed in 5 months • Right First Time without a non-GMP Engineering Run • 5 cGMP runs at 2000 L-scale carried out successfully Proprietary CHO Cell Line
- 15. Jan 2017PepTalk 2017 ViableCellCount(10^6cells/mL) Days 3 L-Scale (n=5) 200 L-Scale (n=1) 2000 L-Scale (n=5) mAbTiter(g/L) Days 3 L-Scale (n=5) 200 L-Scale (n=1) 2000 L-Scale (n=5) Proprietary CHO Cell Line
- 17. Jan 2017PepTalk 2017 Process Transfer from a Client for a mAb • Final cell culture process transferred from client CMO • Gaps were identified in transferred process • Development runs were necessary to establish process conditions • Production bioreactor harvest day chosen based on product quality • Re-optimized process was scaled-up to 2000 L-scale • Successfully implemented strategy of harvesting bioreactor based on product quality target CHO-K1 Cell Line
- 18. Jan 2017PepTalk 2017 CHO-K1 Cell Line ViableCellCount Days 2000 L-Scale 2000 L-Scale 50 L-Scale 15 L-Scale 3 L-Scale 3 L-Scale Glucose Days 2000 L-Scale 2000 L-Scale 50 L-Scale 15 L-Scale 3 L-Scale 3 L-Scale Ammonia Days 2000 L-Scale 2000 L-Scale 50 L-Scale 15 L-Scale 3 L-Scale 3 L-Scale ProductConcentration Days 2000 L-Scale 2000 L-Scale 50 L-Scale 15 L-Scale 3 L-Scale 3 L-Scale %AcidicSpecies Days 50 L-Scale 2000 L-Scale 2000 L-Scale Lactate Days 2000 L-Scale 2000 L-Scale 50 L-Scale 15 L-Scale 3 L-Scale 3 L-Scale ProductQualityAttribute
- 20. Jan 2017PepTalk 2017 Application of Platform Cell Culture Process for 4 mAbs • Expedited process development using an un-optimized platform process • Pre-determined medium-feed combination and bioreactor process parameters • Key objective was speed and consistency rather than quantity of product • Process transferred to manufacturing scale without non- GMP Engineering Run for all 4 mAbs • Gene to IND filing in less than 14 months CHO-S Cell Line
- 21. • HCCF supply runs for purification and analytical method development are typically carried out in 50 L SUB using a stable pool of cells and a platform cell culture process • This approach ensures that compatibility of cell line and process with SUB bags is tested early in development • Wave Cellbags for inoculum expansion • Xcellerex SUB bags for production-stage Jan 2017PepTalk 2017 CHO-S Cell Line Stable Pool HCCF Supply Run (50 L SUB) Clone Selection Process Optimization Platform Process Assessment Process Confirmation (50 L SUB) Scale-up (200 L SUB) Expedited Cell Culture Development Top 3 Clones Top Clone SUB performance evaluation Wave or Seed bioreactor performance evaluation
- 22. Jan 2017PepTalk 2017 ViableCellCount Days 2000 L-Scale 200 L-Scale 50 L-Scale 3 L-Scale 3 L-Scale 3 L-Scale ProductConcentration Days 2000 L-Scale 200 L-Scale 50 L-Scale 3 L-Scale 3 L-Scale 3 L-Scale Glucose Days 2000 L-Scale 200 L-Scale 50 L-Scale 3 L-Scale 3 L-Scale 3 L-Scale Lactate Days 2000 L-Scale 200 L-Scale 50 L-Scale 3 L-Scale 3 L-Scale 3 L-Scale CHO-S Cell Line
- 24. Development and Application of Platform Cell Culture Process for Recombinant Vaccine Proteins • Challenges during process development: • Low productivity & stable-clone selection • Cell viability drop during production-stage • Product cleavage • Challenges during initial scale-up attempts (Cell Line A): • Lack of scalability from glass bioreactor to SUBs » Cell growth and cell viability were significantly divergent • Challenges were overcome; increasing robustness and reproducibility of the platform process (shown by Cell Line B) Jan 2017PepTalk 2017 CHO-DG44 Cell Line
- 25. Jan 2017PepTalk 2017 ViableCellCount Days 200 L-Scale 200 L-Scale 200 L-Scale 200 L-Scale 15 L-Scale 3 L-Scale 3 L-Scale Glucose Days 200 L-Scale 200 L-Scale 200 L-Scale 200 L-Scale 15 L-Scale 3 L-Scale 3 L-Scale CellViability Days 200 L-Scale 200 L-Scale 200 L-Scale 200 L-Scale 15 L-Scale 3 L-Scale 3 L-Scale Lactate Days 200 L-Scale 200 L-Scale 200 L-Scale 200 L-Scale 15 L-Scale 3 L-Scale 3 L-Scale Cell Line A CHO-DG44 Cell Line
- 26. • Observed poor cell growth and drop in cell viability upon scale-up in single-use bioreactor bag • Two key hypotheses • Leachable/extractables • Adsorption of nutrients from basal medium • Steps taken to mitigate both potential risks • Pre-treatment of bioreactor bag (wash) prior to medium addition • Addition of feed media and supplements to overcome depletion of nutrients Jan 2017PepTalk 2017 CHO-DG44 Cell Line
- 27. Jan 2017PepTalk 2017 ViableCellCount Days 100 L-Scale 100 L-Scale 100 L-Scale 3 L-Scale 3 L-Scale 3 L-Scale ProductConcentration Days 100 L-Scale 100 L-Scale 100 L-Scale 3 L-Scale 3 L-Scale 3 L-Scale Glucose Days 100 L-Scale 100 L-Scale 100 L-Scale 3 L-Scale 3 L-Scale 3 L-Scale Lactate Days 100 L-Scale 100 L-Scale 100 L-Scale 3 L-Scale 3 L-Scale 3 L-Scale Cell Line B CHO-DG44 Cell Line
- 28. • Single-use bioreactor scale-up platform has been successfully developed and implemented for a variety of cell lines, cell culture processes and biopharmaceutical products • This single-use bioreactor platform ensures that process scale-up remains smooth and seamless; and cell culture process development can focus on key product quality or titer requirements • KBI Biopharma is uniquely placed with the right knowledge base and experience to serve needs of the biopharmaceutical industry Jan 2017PepTalk 2017
- 29. • Abhinav Shukla • Sigma Mostafa • Cell Culture Process Development • Brian Baker • Lynwel Cunanan • Bryan Howarth • Kathryn Olson • Shaunak Uplekar • Analytical Development & Testing • Jimmy Smedley • Amanda Hoertz’s Group • Nate Oien’s Group Jan 2017PepTalk 2017 • Manufacturing & Manufacturing Sciences • Carnley Norman • Aaron Anders • Ian Bales • Seth Moye • Ronnie Nichols