Simplicity is the key Continuous purification of monoclonal antibodies L. Landric-Burtin Head of Downstream Processing Development, France Integrated Continuous Biomanufacturing Conference, Barcelona, 2013
mabs manufacturing processes have reached a good level of maturity Experience Cell line platforms High titers in cell culture Purification platforms Single use QbD Capacity Demand 2
But our manufacturing processes are still.. COMPLEX! LONG! EXPENSIVE! 3
Equilibration buffer #1 Wash buffer #1 Elution buffer #1 COMPLEX. ph, concentration adjustment, sampling OPERATOR INTERVENTION HOLD TIME OPEN PHASE OPEN PHASE Sampling & Storage OPERATOR INTERVENTION Equilibration buffer #2 Wash buffer #2 Elution buffer #2 HOLD TIME OPEN PHASE Sampling & Storage Bulk Harvest 0.2 µm filtration 0.2 µm filtration Protein A OPERATOR INTERVENTION OPEN PHASE Low ph & Neutralization Equilibration buffer #3 Wash buffer #3 Sampling & Storage OPERATOR INTERVENTION HOLD TIME OPEN PHASE Chrom2 Sampling & Storage OPERATOR INTERVENTION HOLD TIME OPEN PHASE ph, conductivity adjustment Chrom3 Nanofiltration TFF & storage 7 different Contaminations, buffers Human errors 4 7 consecutives, discontinued steps (12 incl. storage)
LONG, DISCONTINUED. Processing time (hr) Around 50 hr to complete the purification process Interruption between steps for concentration, ph or conductivity adjustement and storage. «n» step cannot start before completion of «n-1» 5
EXPENSIVE. 100 /gr Chromatography resin = 30 to 75 % of DSP costs. 6
COMPLEX, LONG, EXPENSIVE... Any solution? 7
SIMPLIFY, SIMPLIFY, SIMPLIFY. SIMPLICITY is key to get a quick, reliable and cheap process. 8
Simplicity & Purification of Mabs? 9
Let s go back to the essentials Equilibration buffer #1 Wash buffer #1 Elution buffer #1 ph, concentration adjustment, sampling Sampling & Storage Equilibration buffer #2 Wash buffer #2 Elution buffer #2 Sampling & Storage Bulk Harvest 0.2 µm filtration 0.2 µm filtration Protein A Low ph & Neutralization Chrom2 Equilibration buffer #3 Wash buffer #3 Sampling & Storage Sampling & Storage ph, conductivity adjustment Chrom3 Nanofiltration TFF & storage 10
Simplicity means NO inter-step ADJUSTMENT. We have selected resins that afford no inter-step adjustment. 11
Simplicity means FEW BUFFERS. We have imagined a buffer strategy composed of only 3 components for the entire process. 12
Simplicity means FEW BUFFERS. 4 buffers in total, ensuring ph and conductivity compatibility between all steps. 13
How simplicity looks like. 14
SIMPLE. Bulk Harvest Equilibration buffer #1 Wash buffer #1 Elution buffer #1 Equilibration buffer #1 Elution buffer #2 Wash buffer #1 Sampling & Storage Low ph Protein A Mixed mode AEX Nanofiltration TFF & storage 4 different buffers 6 steps (7 incl. storage), 4 being continuous. 15
Can a simplified process make the job? Clarified supernatant Protein A Mixed-mode AEX High Molecular Weight species 8.7 % 1.0 % 0.2 % 0.2 % Host Cell Proteins 2.2E+05 ppm 800 ppm 40 ppm 5 ppm Host Cell DNA 3E+06 ppb 2E+04 ppb 10 ppb < 0.1 ppb Endotoxins < 0.05 EU/mL along the process Overall yield 90 % 16
Can a simplified process be a platform? Isoelectric point 9 8 7 6 Project 2 Yield : 89% Project 3 Yield : 92% Project 1 Yield : 93% Project 4 Yield : 94% Project 5 Yield : 86% 5 Successfully evaluated on 5 different projects without optimization. 17
Simplicity advantages 18
SIMPLIFIED PROCESS GAINS reduced our process time: 2X higher per-hour productivity. reduced number of buffers: 30 % cost saving on buffers. 19
SIMPLIFIED PROCESS GAINS Continuous manufacturing with no human handling. Reduced risk of contamination Reduced sampling plan Reduced storage needs Possibility to run the process in a multicycle mode. 20
SIMPLE. Bulk Harvest Equilibration buffer #1 Wash buffer #1 Elution buffer #1 Equilibration buffer #1 Elution buffer #2 Wash buffer #1 Sampling & Storage Low ph Protein A Mixed mode AEX Nanofiltration TFF & storage 4 different buffers 6 steps (7 incl. storage), 4 being continuous. 21
SIMPLE, SMALL and AUTOMATED. Bulk Harvest Equilibration buffer #1 Wash buffer #1 Elution buffer #1 Equilibration buffer #2 Run X Elution buffer #2 Wash buffer #1 Sampling & Storage Run 3 Run 2 Run 1 Protein A Low ph Mixed mode AEX Nanofiltration TFF & storage We have automated the process to run it in multi cycles with small columns. 22
ASAP process gains. Batch Mode Continuous Multi-cycling 66% of resin volume saved for an equivalent batch duration. Up to 97% for > duration. Perfect application for disposable columns. No packing Full use of resin lifetime No storage 23
SIMPLE, SMALL and CHEAP. Batch Mode Continuous Multi-cycling Smaller systems and facilities. Limited staff requested. 24
SIMPLE, SMALL and PRODUCTIVE 14.1 L resin, 4 cycles prot A, (24 hr* to 72h duration) 1 m3 Bulk Harvest Productivity : 2.7 g/l/hr* 1.2 L resin - 47 cycles - 71 hrs duration (*):with shift work Productivity : 10.5 g/l/hr 25
SIMPLE and ADAPTATIVE Adapt your column size to your requirements 50 hr Processing time X hr High product needs, facility constraints: Big columns to process rapidely one batch 0 << 100 /g ~100 /g Cost constraint: Small columns to decrease resin costs and optimize resin lifetime Batch cost 26
CASE STUDY: Continuous purification at lab scale 50 or 100 ml resin volume 70 L of buffers 43 L of supernatant 27
CASE STUDY: Continuous purification at lab scale 43 L of supernatant fully purified continuously in 69h : 45 runs performed automatically. 67g of purified mab collected 93% yield X3 24 ml min Charge 40min X9 (1012)001:10_UV1Col1 X3 24 ml min Charge 40min X9 (1012)001:10_UV1Col2 X3 24 ml min Charge 40min X9 (1012)001:10_UV1Col3 X3 24 ml min Charge 40min X9 (1012)001:10_Logbook mau X3 24 ml min Charge 40min X9 (1012)001:10_UV1Col1 X3 24 ml min Charge 40min X9 (1012)001:10_UV1Col2 X3 24 ml min Charge 40min X9 (1012)001:10_UV1Col3 X3 24 ml min Charge 40min X9 (1012)001:10_Logbook mau 3000 mau mau 2500 3500 3000 2500 3000 2000 2500 2000 2500 2000 1500 2000 1500 1500 1000 1500 1000 1000 1000 500 500 500 500 0 0 0 0 100 200 300 400 min 0 500 1000 1500 2000 2500 3000 3500 4000 min 0 28
SCALE UP and GMP compliance evaluation (On-going) 500 L batch Final target : > 1Kg of purified mab Column < 10L < 10 runs with a final duration of 16h 29
CONCLUSION If you seek for quick, reliable and cheap process, get the superfluous off your process. Starting from this perspective, we have designed - a simplified purification platform, - able to run continuously, - with 4 fold increase in productivity - significant gain on resin cost. 30
and excellent staff Simplicity is the key Continuous purification of monoclonal antibodies Special thanks to: Benoit MOTHES Didier DUTHE Céline HEMET 31
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