Case study: Tech transfer of mammalian cell culture ...§ PID controlpossibilities ... Process scale up from 10L glass to 50L ... scale to production scale using disposable technology

Casestudy:Techtransferofmammaliancellcultureprocessfrom

labscaletoproductionscaleusingdisposabletechnology

DawidSuwałaaa.Mammalian Drug Substance Manufactruring Manager

PolpharmaBiologics

CellWorld 2017, April 26th Page 2Polpharma Biologics property

Techtransferofmammaliancellcultureprocessfromlabscaletoproductionscaleusingdisposabletechnology

AGENDA

Procesdevelopmentinlabscale

TechtransferGAPanalysis– riskevaluation

Processtransferandscale-upto50L

Processscale-upfrom50Lto250L

Final1000LscaleinSingleUseBioreactor


Processdevelopmentchallengesinlabscale

§ Cellline development§ Production mediaselection§ Clonewiththehighest

productivity propertiesselection

§ Feeding andsuplementationstrategy optimization

§ Process parametersoptimization

§ Producttiter andqualityoptimization

§ Threeconsolidation runs inR&Dbefore transfer

§ Dataalignment betweenR&DandProduction Units

Earlystage Latestage Processtransfer


TechTransferGAPanalysis– riskevaluation

Hardwarefeasibilitystudy

Rawmaterialsevaluation

Documentationanddataaligment

§ Proper quality materials usage during process development§ Risk assesment for materials - critical quality parameters analysis§ Raw materials list transfer from R&D to Production

§ Data transfer from R&D to Production – Product Trasfer planand protocols

§ R&D documentation data – clear description of process parametersand introduced changes

§ Criticalequipmentestablishment

§ R&DandProductionequipmentfunctionalitycomparison


KEYELEMENTSOFPROCESSTRANSFER

Bioreactorscharacterization

§ Hardwarefeasibility study

§ Rawmaterialsevaluation

§ Documentation anddataalignmentRisk evaluation

§ Mixing studies

§ Masstransferdetermination (kLa)

§ Powerinputdetermination(empiricaldetermination)

§ CFDcharacterization

TechTransferGAPanalysis– riskevaluation


VS

Laboratory andProduction scale – how toalign?

Two stepapproach

Processscaleupfrom10Lglassto50LSingleUseBioreactor



Equipment functional performance

Newculture environment– glass toplasticexchangeandsingleuse bioreactor bag internal layer filminfluenceonculture performance

Process optimization – operational parameters adaptationtonew environment

GMPcell banks production andcharacterization;introductiontoroutine use

Whatisthecase?

1

2

3

4

Firststep– process transfer


What is thecase?

§ Vessel andstirrer geometry§ Material contact forcell culture

§ Gassing pathways

§ Spargers types§ PIDcontrol possibilities

Equipment functionalperformance1



What is thecase?

0.00

1.00

2.00

3.00

4.00

5.00

6.00

7.00

0.0010.0020.0030.0040.0050.0060.0070.0080.0090.00100.00

0 24 48 72 96 120 144

VCC[0

6viablecells/ml]

ViabilityofCulture

[%]

ElapsedTime[h]


§ Gassing pathways



§ Cellsviability andculture density§ Productivity

§ Productprofile(chargévariants,aggregation,etc.)

2Newculture environment–SUBinternal layer film

influenceonculture performance



What is thecase?


§ Gassing pathways





2

§ Gassingstrategy

§ Spargingstrategy§ PIDcontrolstartegy

§ SingleUseBioreactorprojectcustomization

Process optimization 3

Newculture environment–SUBinternal layer film

influenceonculture performance



What is thecase?



Newculture environment–SUBinternal layer film

influenceonculture performance2

§ Cellbanks characterization

§ Cellbanks quality§ Newcell banks influenceon

process performance

MCB&WCBproductionandintroductiontoroutine use

4

§ Gassingstrategy

§ Spargingstrategy§ PIDcontrolstartegy

§ SingleUseBioreactorprojectcustomization

Process optimization 3


§ Gassing pathways





Processscaleupfrom50Lto250LSingleUseBioreactor

50LSUBTechnicalTrial Dataanalysis 250LSUBTechnicalTrial

§ Riskevaluation§ R&DDataevaluation§ Techtransferprotocols

establishment§ Technicaltrialsperformance

andexecution§ Technicaltrialdatacollection

§ Technicaltrialdataevaluation

§ Analyticaldataevaluation§ Processmismatches/

unexpectedeventsrootcauseinvestigations

§ Transferreportpreparationincludingconclusionstoachieveddataandrecommendationsfornexttrials

§ TechnicaltrialrepetitionifneededorprocessoptimizationinR&Dformoredatacollection

§ Preliminaryprocessparametersestablishment

§ Techtransferprotocolsestablishment

§ Technicaltrialsexecution§ Technicaltrialdatacollection

CASE:WCBqualityinfluenceonprocessperformance

Bioreactors characterization is akey element

Secondstep– process scale-up


250LSUBTechnicalTrial 1000LSUBTechnicalTrial

§ Preliminaryprocessparametersestablishment

§ Techtransferprotocolsestablishment

§ Technicaltrialsexecution§ Technicaltrialdatacollection

§ Technicaltrialdataevaluation

§ Analyticaldataevaluation§ Processmismatches/

unexpectedeventsrootcauseinvestigations

§ Transferreportpreparationincludingconclusionstoachieveddataandrecommendationsfornexttrials

§ ProcessPrametersestablishment

§ Engeeneringbatchesperformanceandexecution

§ Processoptimization§ IPCstrategyestablishment


Bioreactors characterization is akey element

Secondstep– process scale-upDataanalysis


Parameter Specification 1000LBatch Consolidation runs*

mAb content 18-22mg/ml 21,3mg/ml 19,8÷20,3mg/ml

Purity (SEC)

HMWI≤0,2% N/A N/A

dimers ≤1,0% 0,3% 0,2÷ 0,4%

Purity ≥97,0% 99,0% 98,4÷ 98,9%

Purity (CE) ≥92,0% 97,7% 98,0÷ 98,3%

CEX

Sumofacidic peaks ≤18,0% 13,4% 14,8÷ 15,8%

Main Peak ≥70% 72,6% 70,8÷71,6%NA 14,0% 13,213,613,4

HCP ≤100ppm 20ppm 9÷ 20ppm

DNA ≤33ppb Below LOD<0.075 0,1÷ 0,3ppb

ProteinA ≤10ppm 0,1ppm 0,2ppm

Glycan mappingG0F:38,8-71,1% 48,2% 54,1÷ 53,9%

SumofG1F,G1F',G2F:32-58% 45,9% 38,3÷ 38,7%

Polisorbate 80content 0,1- 0,3mg/ml 0,2mg/ml N/A

Potency 80-125% 103% 100÷ 102%

Endotoxin ≤7,3IU/ml <0,125IU/ml N/A

Bioburden ≤1CFU/10mL <1CFU/10mL N/A

pH 5,5÷ 5,9 5,7 5,7÷ 6,1

Osmolarity ≥240mosmol/kg 296mosmol/kg 281÷ 291mosmol/kg

*Minandmaxresults fromtheconsolidation round 3(Run1,2and3)



Riskevaluationandbioreactorscharacterizationarekeyforprocesstransferandscale-up

Twostepapproachgavepositiveresultofprocessoptimizationindisposablesystems

Finalscale-updonewithproductqialityspecificparametrspreservationafterallproblemweresolvedinpreviousactivities

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SUMMARY


TakeHomeMessage

To avoid mismatches between lab scale and production platform during technology transfer it is needed to predict production performance at early stage of lab scale process description and hardware setup. Wellknown bioreactors characteristic and risk evaluation are key elements

during process transfer and scale-up.




Thank you for your attention

Documents

Case study: Tech transfer of mammalian cell culture ...§ PID controlpossibilities ... Process scale up from 10L glass to 50L ... scale to production scale using disposable technology