a case study on the application of disposable technologies
TRANSCRIPT
A Case Study on the Application of Disposable Technologies in cGMP Manufacturing Processes for a Therapeutic Antibody
Martin WrankmoreLonza Biologics (UK)
Presented at:Biologicals Manufacturing SummitFebruary 23rd - 24th 2005London, UK
Slide 2
Presentation Outline
■ Why disposables?■ Traditional process overview■ Applications
■ Tank Liners■ Bio-Process Containers – BPCs (Bags)■ Tube Welding■ Disposable Bioreactors■ Harvest Systems■ Capsule Filtrations with BPCs
■ New process overview■ Benefits & Challenges■ Acknowledgements
Slide 3
What Does Lonza Do?
LonzaGroupSwiss based life sciences driven companyCustom manufacturer of active chemical ingredients, intermediates and biotechnology solutions to the pharmaceutical and agrochemical industries
Lonza BiologicsCustom manufacturer of therapeutic monoclonal antibodies and recombinant proteins from mammalian cell cultures making bulk APIOperations range from cell line construction and process development to large scale manufacturing and regulatory supportSites in Slough (UK) & Portsmouth (NH, USA)
Slide 4
Why disposables?
Drivers for useIncreased flexibility of operations for wide range of processesMinimisation of plant validationReduced unit operation timesAbility to change processes quickly
AdvantagesNo cleaning requirementsNo sterilisation procedures needed before useFast preparation (often none) & ease of useNo product carryover/contamination risk
ConsiderationsCostsExtractables issuesReliance on external supply chain
Slide 5
Traditional Mammalian Cell GMP Process
Ampoule
FlaskFlasks
Ampoule
FlaskFlasks
ProductionFermenter
InoculumFermenter
InoculumFermenter
Feeds
HoldingTank
MixingTank
Media PreparationConcentration
HoldingTank
HoldingTank
SterileNalgene
ClarificationFilters
IntermediateFiltration
5±3 °C
Protein AChromatography
In ProcessFiltration
2nd StageChromatography
In ProcessFiltration
Concentration /Dia-filtration
In ProcessFiltration
In ProcessFiltration
Virus ReductionFiltration
3rd StageChromatography
Concentration /Dia-filtration
In ProcessFiltration
In ProcessFiltration
Concentration /Dia-filtration
In ProcessFiltration
FinalFiltration
5±3 °C
ProductNalgenes
HoldingTank
MixingTank
Buffer Preparation
Slide 6
Applications - Tank Liners & BPCs
HoldingTank
MixingTank
Buffer Preparation
ProductionFermenter
InoculumFermenter
InoculumFermenter
Feeds
HoldingTank
MixingTank
Media Preparation
Ampoule
FlaskFlasks
Concentration
HoldingTank
HoldingTank
SterileNalgene
ClarificationFilters
IntermediateFiltration
5±3 °C
Protein AChromatography
In ProcessFiltration
2nd StageChromatography
In ProcessFiltration
Concentration /Dia-filtration
In ProcessFiltration
In ProcessFiltration
Virus ReductionFiltration
3rd StageChromatography
Concentration /Dia-filtration
In ProcessFiltration
In ProcessFiltration
Concentration /Dia-filtration
In ProcessFiltration
FinalFiltration
5±3 °C
ProductNalgenes
Slide 7
Applications - Tank Liners
Replaces medium and buffer make-up tanksFlexible, pre-sterilised open bagsMounted into rigid non-contact reusable containersWFI, Stirrer assembly & additions made through top of linerMixed solution pumped outUsed currently for cell culture medium and feed preparation – planned to extend to buffer preparation
Slide 8
Applications - Bio-Process Containers (BPCs)
Replaces pre-cleaned steel tanks and glass bottlesFlexible, pre-sterilised plastic containers50mL to 500L scaleFilled bags stored in re-usable containersUsed for medium, feeds, buffers, WFI & sample collection
Slide 9
Tank Liners & BPCs - Advantages
No pre-use cleaning or sterilising of equipment required nor post-use cleaningSignificant reduction in both duration and resource requirementsAbility to be much more flexible in volumes preparedBPCs easier to use on balances for monitoring amounts used/deliveredStorage of filled BPCs much easier and more flexibleHas enabled more complex feeding regimes to be usedBPCs with integral filters have replaced separate filtration units and multiple stagesCreation of a new or expansion of existing media or buffer preparation facility now easier and quickerNo need to prepare sampling devices in VLFC
Slide 10
Tank Liners & BPCs – Challenges
Tank liners can be easily split by impellerImpeller system must be cleanedDifferent films used in BPCs by different suppliers and problems when film changes are madeSupply chain issues can cause problems especially if wide range of BPCs usedIncorrect manufacture of BPCs (wrong filters, wrong fittings or in incorrect positions)Care with not letting bag out of containerCost of BPCsBag splits – never happened so far
Slide 11
Applications - Tube Welding & Disposable Bioreactors
HoldingTank
MixingTank
Buffer Preparation
ProductionFermenter
InoculumFermenter
InoculumFermenter
Feeds
HoldingTank
MixingTank
Media Preparation
Ampoule
FlaskFlasks
Concentration
HoldingTank
HoldingTank
SterileNalgene
ClarificationFilters
IntermediateFiltration
5±3 °C
Protein AChromatography
In ProcessFiltration
2nd StageChromatography
In ProcessFiltration
Concentration /Dia-filtration
In ProcessFiltration
In ProcessFiltration
Virus ReductionFiltration
3rd StageChromatography
Concentration /Dia-filtration
In ProcessFiltration
In ProcessFiltration
Concentration /Dia-filtration
In ProcessFiltration
FinalFiltration
5±3 °C
ProductNalgenes
Slide 12
Applications - Tube Welding
Replaces local SIP stainless steel connectionsUse C-flex tubingHot blade (≈200°C) used to cut tubeJaws re-align tubes such that appropriate connection is madeBlade removed and tubing anneals to form sterile sealUsed in fermentation and planned for use in purification
Slide 13
Tube Welding
AdvantagesTube welder has provided considerable savings
Old SIP system took > 1½ hTube welder takes ≈ 2 min
Far greater flexibility – more connections possible
ChallengesEnsuring weld is good before proceedingBlades expensiveIncreased use of non-permanent tubing around plantMay increase number of aseptic operations
Slide 14
Applications – Disposable Bioreactors
Replaces two stages of inoculum fermenter hardwareUses disposable bags (each up to 50L capacity)Temperature controlledBag pressurised by 5% CO2 in air (gives pH & DO control)Mixed by rocking motion of bedUsed like a large shake flask
Slide 15
Disposable Bioreactors – Comparability
1.0E+5
1.0E+6
1.0E+7
0 20 40 60 80 100 120 140
Elapsed Time (Hours)
Viab
le c
ell c
once
ntra
tion
(/mL)
S200#2
S200#1
S200#4
S200#5
S200#6
AirliftInoculum
Slide 16
Disposable Bioreactors
AdvantagesPreparation times reduced from 3 or 4 days to less than oneCan be used as production system with multiple bags in parallelMore flexibility in operational volumesContamination rate very low
ChallengesGas filters wetting outCare over cell culture transfersIncreased 5% CO2 in air gas usageTemperature validation
Slide 17
Applications - Harvest Systems
HoldingTank
MixingTank
Buffer Preparation
ProductionFermenter
InoculumFermenter
InoculumFermenter
Feeds
HoldingTank
MixingTank
Media Preparation
Ampoule
FlaskFlasks
Concentration
HoldingTank
HoldingTank
SterileNalgene
ClarificationFilters
IntermediateFiltration
5±3 °C
Protein AChromatography
In ProcessFiltration
2nd StageChromatography
In ProcessFiltration
Concentration /Dia-filtration
In ProcessFiltration
In ProcessFiltration
Virus ReductionFiltration
3rd StageChromatography
Concentration /Dia-filtration
In ProcessFiltration
In ProcessFiltration
Concentration /Dia-filtration
In ProcessFiltration
FinalFiltration
5±3 °C
ProductNalgenes
Slide 18
Applications - Harvest Systems
Replaces multi stage stainless steel harvest systemUses 5 stage filtration processAll filters as disposable capsulesConcentration step omittedUsed for harvest of 200L fed batch fermentations
Slide 19
Harvest Systems
AdvantagesRemoves need for several days of preparation prior to harvestMore flexibility in harvest timingGreater capability to increase filter area if unexpected blocking occursProcess can be visually monitored more easily
ChallengesDiluted purification process streamConsumable costs high
Slide 20
Applications - Capsule Filters with BPCs
HoldingTank
MixingTank
Buffer Preparation
ProductionFermenter
InoculumFermenter
InoculumFermenter
Feeds
HoldingTank
MixingTank
Media Preparation
Ampoule
FlaskFlasks
Concentration
HoldingTank
HoldingTank
SterileNalgene
ClarificationFilters
IntermediateFiltration
5±3 °C
Protein AChromatography
In ProcessFiltration
2nd StageChromatography
In ProcessFiltration
Concentration /Dia-filtration
In ProcessFiltration
In ProcessFiltration
Virus ReductionFiltration
3rd StageChromatography
Concentration /Dia-filtration
In ProcessFiltration
In ProcessFiltration
Concentration /Dia-filtration
In ProcessFiltration
FinalFiltration
5±3 °C
ProductNalgenes
Slide 21
Applications - Capsule Filters with BPCs
Replaces cartridge elements and stainless steel housingsAvailable in wide range of filter areas and capacitiesLinked to BPCsUsed to perform in-process filtrations of product streamIn Purification on each filtration step:
reduced three process steps to onesaved over 2h duration
Slide 22
Process Overview – Original Process
Ampoule
FlaskFlasks
Ampoule
FlaskFlasks
ProductionFermenter
InoculumFermenter
InoculumFermenter
Feeds
HoldingTank
MixingTank
Media PreparationConcentration
HoldingTank
HoldingTank
SterileNalgene
ClarificationFilters
IntermediateFiltration
5±3 °C
Protein AChromatography
In ProcessFiltration
2nd StageChromatography
In ProcessFiltration
Concentration /Dia-filtration
In ProcessFiltration
In ProcessFiltration
Virus ReductionFiltration
3rd StageChromatography
Concentration /Dia-filtration
In ProcessFiltration
In ProcessFiltration
Concentration /Dia-filtration
In ProcessFiltration
FinalFiltration
5±3 °C
ProductNalgenes
HoldingTank
MixingTank
Buffer Preparation
Slide 23
Process Overview – Revised Process
5±3 °C
Buffer Preparation
BPC
HarvestFiltration
BPC
Media Preparation
BPC
ProductionFermenter
Wave Bioreactor
Wave Bioreactor
BPC
Feeds
Ampoule
FlaskFlasks
FinalFiltration
5±3 °C
ProductNalgenes
3rd StageChromatography
In ProcessFiltration
Concentration /Dia-filtration
In ProcessFiltration
Virus ReductionFiltration
Concentration /Dia-filtration
In ProcessFiltration
In ProcessFiltration
In ProcessFiltration
In ProcessFiltration
Concentration /Dia-filtration
In ProcessFiltration
Slide 24
Summary of Benefits
Process simplification opportunitiesIncreased process robustnessSignificantly reduced onsite preparation workReduction in reliance on utilities and servicesMore flexibilityMinimises risk of product cross contaminationExtensive use of gamma sterilisationPotential for completely disposable process especially upstream – disposable downstream systems catching up rapidly
Slide 25
Ongoing Challenges
Reliance on suppliers for both delivery and flexibility in design / customisationOperating disposables under GMPKeeping up with rapid changes in available technologiesBalancing increased consumable costs against potential/realised savings in time and resource costsValidation issues – what can be taken direct from supplier and what needs completion in house
Slide 26
Acknowledgements
Lonza Biologics:Co-authors
Jeremy M Tong, Mark S Kettel, Edward M Perry, David J Pain, David M Valentine, Ashley J Westlake & Michael Brown
ManufacturingContinuous ImprovementPilot & Scale-Up SupportQA & QCDocumentation
Various Disposables Suppliers
Thank you for your attention
Questions?