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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.
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Simplicity is the key Continuous purification of monoclonal antibodies L. Landric-Burtin Head of Downstream Processing Development, France Integrated Continuous BiomanufacturingConference, Barcelona, 2013
mAbsmanufacturingprocesses have reached a good level of maturity Experience Cell line platforms High titers in cell culture Purification platforms Single use QbD Capacity ≥ Demand
But ourmanufacturingprocesses are still.. LONG ! EXPENSIVE ! COMPLEX !
COMPLEX. OPERATOR INTERVENTION Equilibration buffer #1 Equilibration buffer #2 Wash buffer #1 Wash buffer #2 OPERATOR INTERVENTION Elution buffer #1 Elution buffer #2 HOLD TIME HOLD TIME pH, concentration adjustment, sampling OPEN PHASE OPEN PHASE Sampling & Storage Sampling & Storage OPEN PHASE BulkHarvest 0.2 µm filtration 0.2 µm filtration Protein A Chrom2 LowpH & Neutralization OPERATOR INTERVENTION Equilibration buffer #3 OPERATOR INTERVENTION OPERATOR INTERVENTION HOLD TIME Wash buffer #3 HOLD TIME OPEN PHASE Sampling & Storage Sampling & Storage OPEN PHASE OPEN PHASE pH, conductivityadjustment Nanofiltration TFF & storage Chrom3 ≥ 7 different buffers 7 consecutives, discontinuedsteps (12 incl. storage) Contaminations, Humanerrors
LONG, DISCONTINUED. Around 50 hr to completethe purification process Processing time (hr) « n » stepcannotstartbeforecompletion of « n-1 » Interruption betweensteps for concentration, pH or conductivityadjustement and storage.
EXPENSIVE. • ≥ 100 €/gr • Chromatographyresin = 30 to 75 % of DSP costs.
COMPLEX, LONG, EXPENSIVE... Any solution ?
SIMPLIFY, SIMPLIFY, SIMPLIFY. → SIMPLICITY is key to get a quick, reliable and cheap process.
Simplicity • & • Purification of Mabs?
Let’s go back to the essentials Equilibration buffer #1 Equilibration buffer #2 Wash buffer #1 Wash buffer #2 Elution buffer #2 Elution buffer #1 pH, concentration adjustment, sampling Sampling & Storage Sampling & Storage BulkHarvest 0.2 µm filtration 0.2 µm filtration Protein A Chrom2 LowpH & Neutralization Equilibration buffer #3 Wash buffer #3 Sampling & Storage Sampling & Storage pH, conductivityadjustment TFF & storage Nanofiltration Chrom3
Simplicitymeans NO inter-step ADJUSTMENT. →Wehave selectedresinsthatafford no inter-stepadjustment.
Simplicitymeans FEW BUFFERS. →We have imagined a buffer strategycomposed of only 3 components for the entireprocess.
Simplicitymeans FEW BUFFERS. →4 buffers in total, ensuring pH and conductivity compatibility between all steps.
SIMPLE. BulkHarvest Equilibration buffer #1 Wash buffer #1 Elution buffer #1 Equilibration buffer #1 Elution buffer #2 Wash buffer #1 Sampling & Storage Low pH TFF & storage Nanofiltration Protein A AEX Mixed mode • 4 different buffers • 6 steps (7 incl. storage), 4 beingcontinuous.
Can a simplifiedprocessmakethe job ? Endotoxins < 0.05 EU/mLalong the process Overallyield ≥ 90 %
Can a simplifiedprocessbe a platform ? Project 2 Yield : 89% Isoelectric point Project 4 Yield : 94% Project 3 Yield : 92% Project 1 Yield : 93% Project 5 Yield : 86% → Successfullyevaluated on 5 differentprojectswithoutoptimization.
SIMPLIFIED PROCESS GAINS → reducedourprocess time: 2X higherper-hourproductivity. • → reducednumberof buffers: • 30 % costsaving on buffers.
SIMPLIFIED PROCESS GAINS • →Continuous manufacturing with no human handling. • Reducedrisk of contamination • Reducedsampling plan • Reducedstorageneeds • Possibility to run the process in a multicycle mode.
SIMPLE. BulkHarvest Equilibration buffer #1 Wash buffer #1 Elution buffer #1 Equilibration buffer #1 Elution buffer #2 Wash buffer #1 Sampling & Storage Low pH TFF & storage Nanofiltration Protein A AEX Mixed mode • 4 different buffers • 6 steps (7 incl. storage), 4 beingcontinuous.
SIMPLE, SMALL and AUTOMATED. BulkHarvest Equilibration buffer #1 Wash buffer #1 Elution buffer #1 Equilibration buffer #2 Elution buffer #2 Sampling & Storage Wash buffer #1 Run X Run 3 Run 2 Run 1 Low pH Mixed mode Protein A AEX TFF & storage Nanofiltration →We have automated the process to runit in multi cycles withsmallcolumns.
ASAP process gains. • 66% of resin volume saved for an equivalent batch duration. • Up to 97% for > duration. • Perfectapplication for disposablecolumns. • No packing • Full use of resinlifetime • No storage Continuous Multi-cycling Batch Mode
SIMPLE, SMALL and CHEAP. • Smallersystems and facilities. • Limited staff requested. Batch Mode Continuous Multi-cycling
SIMPLE, SMALL and PRODUCTIVE 14.1 L resin, 4 cycles prot A, (24 hr* to 72h duration) Productivity : 2.7 g/L/hr* 1 m3 BulkHarvest (*):with shift work 1.2 L resin - 47 cycles - 71 hrsduration Productivity : 10.5 g/L/hr
SIMPLE and ADAPTATIVE → Adaptyourcolumn size to yourrequirements Processing time 50 hr X hr High productneeds, facilityconstraints: → Bigcolumns to processrapidely one batch Batch cost 0 << 100€/g ~100€/g Costconstraint: → Small columns to decreaseresincosts and optimizeresinlifetime
CASE STUDY:Continuous purification atlabscale 70 L of buffers 50 or 100 mLresin volume 43 L of supernatant
CASE STUDY:Continuous purification atlabscale • 43 L of supernatantfullypurifiedcontinuously in 69h : • 45 runsperformedautomatically. • 67g of purifiedmAbcollected • 93% yield
SCALE UP and GMP complianceevaluation (On-going) • 500 L batch • Final target : > 1Kg of purifiedmAb • Column < 10L • < 10 runswith a final duration of 16h
CONCLUSION • →If youseekfor quick, reliable and cheap process, get the superfluous off yourprocess. • → Startingfromthis perspective, we have designed • a simplified purification platform, • able to runcontinuously, • with 4 foldincrease in productivity • significant gain on resincost.
and excellent staff • Simplicity is the key • Continuous purification of monoclonal antibodies Specialthanks to: Benoit MOTHES Didier DUTHE Céline HEMET