a case study on the application of disposable technologies...

A Case Study on the Application of Disposable Technologies in cGMP Manufacturing Processes for a Therapeutic Antibody.

Jeremy M Tong (presenting),Mark S Kettel, Edward M Perry,David J Pain, David M Valentine,Ashley J Westlake, Martin Wrankmore& Michael Brown

Lonza Biologics (UK)

ACS 227th National Meeting2nd Apr 2004

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

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

Ampoule

FlaskFlasks

Ampoule

FlaskFlasks

Traditional Mammalian Cell GMP Process

ProductionFermenter

InoculumFermenter

InoculumFermenter

Feeds

HoldingTank

MixingTank

Media PreparationConcentration

HoldingTank

HoldingTank

SterileNalgene

ClarificationFilters

IntermediateFiltration

5±3 °C

Protein AChromatography

In ProcessFiltration

2nd StageChromatography


Concentration /Dia-filtration



Virus ReductionFiltration

3rd StageChromatography






FinalFiltration

5±3 °C

ProductNalgenes

HoldingTank

MixingTank

Buffer Preparation

Slide 6

HoldingTank

MixingTank

Buffer Preparation

ProductionFermenter

InoculumFermenter

InoculumFermenter

Feeds

HoldingTank

MixingTank

Media Preparation

Ampoule

FlaskFlasks

Applications - Tank Liners

Concentration

HoldingTank

HoldingTank

SterileNalgene



5±3 °C















FinalFiltration

5±3 °C

ProductNalgenes

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

Applications - Tank Liners

Slide 8

Applications - Bio-Process Containers (BPCs)

Replaces pre-cleaned steel tanks and glass bottlesFlexible, pre-sterilised plastic containers20 to 500L scaleFilled bags stored in re-usable containersUsed for medium, feeds, buffers & WFI

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 quicker

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 (wrong filters, wrong fittings or in incorrect positions)Care with not letting bag out of containerBPCs are quite costlyBag splits – never happened so far

HoldingTank

MixingTank

Buffer Preparation

ProductionFermenter

InoculumFermenter

InoculumFermenter

Feeds

HoldingTank

MixingTank

Media Preparation

Ampoule

FlaskFlasks

Applications - Tube Welding

Concentration

HoldingTank

HoldingTank

SterileNalgene



5±3 °C















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

Tube Welder

AdvantagesTube welder has provided considerable savings

Old SIP system took > 1½ h; generated chart trace that required reviewTube welder takes ≈ 2 min and generates no data for review

Far greater flexibility – more connections possibleChallenges

Ensuring weld is good before proceedingBlades expensiveIncreased use of non-permanent tubing around plant

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

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

HoldingTank

MixingTank

Buffer Preparation

ProductionFermenter

InoculumFermenter

InoculumFermenter

Feeds

HoldingTank

MixingTank

Media Preparation

Ampoule

FlaskFlasks

Applications - Harvest Systems

Concentration

HoldingTank

HoldingTank

SterileNalgene



5±3 °C















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 fermentation

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

HoldingTank

MixingTank

Buffer Preparation

ProductionFermenter

InoculumFermenter

InoculumFermenter

Feeds

HoldingTank

MixingTank

Media Preparation

Ampoule

FlaskFlasks

Applications - Capsule Filters with BPCs

Concentration

HoldingTank

HoldingTank

SterileNalgene



5±3 °C















FinalFiltration

5±3 °C

ProductNalgenes

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



5±3 °C















FinalFiltration

5±3 °C

ProductNalgenes

HoldingTank

MixingTank

Buffer Preparation

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













Process Overview – Revised Process

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

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-authorsManufacturingContinuous ImprovementPilot & Scale-Up SupportQA & QCDocumentation

Various Disposables Suppliers

Thank you for your attention

Questions?

a case study on the application of disposable technologies...

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