Edited by

Ganapathy Subramanian

Biopharmaceutical Production Technology

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Edited by Ganapathy Subramanian

Biopharmaceutical Production Technology

Volume 1

The Editor

Dr. Ganapathy Subramanian44 Oaken GroveMaidenheadBerkshire SL6 6HHUnited Kingdom

All books published by Wiley-VCH are carefully produced. Nevertheless, authors, editors, and publisher do not warrant the information contained in these books, including this book, to be free of errors. Readers are advised to keep in mind that statements, data, illustrations, procedural details or other items may inadvertently be inaccurate.

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© 2012 Wiley-VCH Verlag & Co. KGaA, Boschstr. 12, 69469 Weinheim, Germany

All rights reserved (including those of translation into other languages). No part of this book may be reproduced in any form – by photoprinting, microfilm, or any other means – nor transmitted or translated into a machine language without written permission from the publishers. Registered names, trademarks, etc. used in this book, even when not specifically marked as such, are not to be considered unprotected by law.

Composition Toppan Best-set Premedia Limited, Hong KongPrinting and Binding Markono Print Media Pte Ltd, SingaporeCover Design Adam-Design, Weinheim

Print ISBN: 978-3-527-33029-4ePDF ISBN: 978-3-527-65312-6ePub ISBN: 978-3-527-65311-9mobi ISBN: 978-3-527-65310-2oBook ISBN: 978-3-527-65309-6

Printed in SingaporePrinted on acid-free paper

V

Contents

Preface XXIII ListofContributors XXV

Volume1

PartOne UpstreamTechnologies 1

1 StrategiesforPlasmidDNAProductioninEscherichia coli 3EvaBrand,KathrinRalla,andPeterNeubauer

1.1 Introduction 31.2 RequirementsforaPlasmidDNAProductionProcess 41.3 StructureofaDNAVaccineProductionProcess 61.4 ChoiceofAntigen 71.5 VectorDNAConstruct 81.5.1 PopularAmplificationSystems 81.5.2 IntrinsicFactors 91.6 HostStrains 111.6.1 endAandrecA  121.6.2 relA  121.6.3 NucleosidePathway 141.6.4 gyrA  151.6.5 StrainsforProductionProcesses 151.7 CultivationMediumandProcessConditions 161.8 Lysis/ExtractionofPlasmidDNA 191.9 Purification 201.9.1 ClarificationoftheLysateandIntermediatePurification 211.9.2 PurificationbyChromatography 231.9.2.1 Anion-ExchangeChromatography 231.9.2.2 HydrophobicInteractionChromatography 241.9.2.3 GelFiltration 241.9.2.4 MembraneChromatography 241.9.2.5 ChromatographyonPorousMonolithicSupports 25

VI Contents

1.10 Formulation 261.10.1 Lipoplexes 271.10.2 Polyplexes 271.10.3 InorganicNanoparticles 281.11 Conclusions 28

References 28

2 AdvancesinProteinProductionTechnologies 43LindaH.L.LuaandYapPangChuan

2.1 Introduction 432.2 GlycoengineeringforHomogenousHuman-LikeGlycoproteins 452.3 BacteriaasProteinFactories 472.4 MammalianCellTechnology 502.5 YeastProteinProduction 532.6 Baculovirus–InsectCellTechnology 552.7 TransgenicAnimalProteinProduction 572.8 PlantMolecularFarming 592.9 Cell-FreeProteinProduction 622.10 FutureProspects 65

References 66

PartTwo ProteinRecovery 79

3 ReleasingBiopharmaceuticalProductsfromCells 81AntonP.J.Middelberg

3.1 Introduction 813.2 CellStructureandStrategiesforDisruption 833.3 CellMechanicalStrength 853.4 Homogenization 893.4.1 Mechanisms 903.4.2 Modeling 913.5 BeadMilling 953.5.1 Modeling 963.6 ChemicalTreatment 983.7 CellularDebris 1003.7.1 Modeling 1023.8 Conclusions 103

References 104

4 ContinuousChromatography(MulticolumnCountercurrentSolventGradientPurification)forProteinPurification 107GuidoStröhlein,ThomasMüller-Späth,andLarsAumann

4.1 Introduction 1074.1.1 OverviewoftheBiopharmaceuticalMarket 1074.1.2 OverviewofPurificationofBiopharmaceuticals 1084.1.3 IntroductiontoContinuousChromatographicProcesses 108

Contents VII

4.2 OverviewofContinuousChromatographicProcesses 1104.2.1 SMBandItsDerivatives 1104.2.1.1 ApplicationsofSMBinthePharmaceuticalIndustry:

SmallMolecules 1114.2.1.2 LimitationsofSMB 1124.2.2 MCSGPGoesBeyondSMBandMakesContinuousChromatography

PossibleforBioseparations 1124.3 PrinciplesofMCSGP 1134.3.1 TasksinBatchChromatogram 1134.3.1.1 GenericPurificationProblem 1144.3.2 Six-ColumnMCSGPPrinciple 1154.3.3 Three-ColumnMCSGPPrinciple 1154.3.4 Four-ColumnMCSGPwithSeparateCIPPosition 1164.3.5 Four-ColumnMCSGPwithaSeparatePositionforContinuous

Feed 1184.3.6 MCSGPProcessforSeparationswithMoreThanThreeFractions 1194.4 ApplicationExamplesofMCSGP 1204.4.1 PolypeptidePurificationwithReversed-PhaseChromatography 1204.4.2 mAbChargeVariantSeparation 1254.4.3 mAbCaptureandPolishfromSupernatant 1274.4.4 Size-ExclusionChromatographicPurificationwithMCSGP 1294.5 EnablingFeaturesandEconomicImpactofMCSGP 1344.6 Annex1:ChromatographicProcessDecisionTree 135

References 136

5 Virus-LikeParticleBioprocessing 139YapPangChuan,LindaH.L.Lua,andAntonP.J.Middelberg

5.1 Introduction 1395.2 UpstreamProcessing 1435.2.1 IntracellularExpressionandAssembly 1435.2.2 Cell-FreeApproaches 1475.3 DownstreamProcessing 1475.3.1 GardasilDownstreamProcessing 1485.3.2 VLPAggregation 1495.3.3 PurificationofCell-AssembledVLPs 1505.3.4 PurificationforIn VitroAssembly 1525.4 Analysis 1545.5 Conclusions 1575.6 Nomenclature 158 Acknowledgments 158

References 158

6 TherapeuticProteinStabilityandFormulation 165RobertFalconar

6.1 Introduction 1656.2 ProteinStability 167

VIII Contents

6.2.1 StructuralStability 1676.2.2 ThermalStability 1686.2.3 Chaotropes,Solvents,andpH 1686.2.4 Shear 1696.2.5 Freezing 1696.2.6 Drying 1706.2.7 Air–LiquidandSolid–LiquidInterfaces 1706.2.8 ChemicalStability 1716.2.9 Precipitation,Aggregation,andFibrilFormation 1736.2.10 Leachables 1746.3 FormulationandMaterials 1756.3.1 LiquidFormulations 1756.3.2 pH 1766.3.3 AminoAcidsandOtherOrganicBuffers 1776.3.4 SugarsandPolyols 1776.3.5 Salts 1776.3.6 Surfactants 1786.3.7 SpecificBinding 1786.3.8 ChelatingAgents 1786.3.9 RedoxPotential 1796.3.10 ContainersandClosures 1796.3.11 FrozenFormulations 1796.3.12 Freeze-DriedFormulations 1806.4 ScreeningMethods 1856.4.1 DSC 1856.4.2 ThermalScanningwithSpectroscopicDetectionofProtein

Unfolding 1876.5 AcceleratedandLong-TermStabilityTesting 1886.5.1 RegulatoryPerspective 1886.5.2 AcceleratedStabilityTesting 1896.6 AnalyticalTechniquesforStabilityTesting 1896.6.1 Cell-BasedBioassaysandIn VitroBindingAssays 1906.6.2 High-PerformanceLiquidChromatographyandCapillaryZone

Electrophoresis 1916.6.3 MassSpectrometry-BasedAnalysis 1926.6.4 DetectionofProteinAggregates 1926.6.5 CrudeAnalyticalAssays:PAGE,IEF,Blotting,FTIR,CD,andUV

Fluorescence 1936.7 Conclusions 194

References 195

7 ProductionofPEGylatedProteins 199ConanJ.FeeandVinodB.Damodaran

7.1 Introduction 1997.2 GeneralConsiderations 200

Contents IX

7.2.1 EfficiencyofPEGConjugation 2007.2.2 ControlofPositionalIsomerism 2017.2.3 ControloftheNumberofPEGAdducts 2027.2.4 PurificationofTargetProducts 2037.3 PEGylationChemistry 2047.3.1 AmineConjugation 2047.3.2 ThiolConjugation 2067.3.3 OxidizedCarbohydrateorN-TerminalConjugation 2087.3.4 Transglutaminase-MediatedEnzymaticConjugation 2087.3.5 MiscellaneousConjugationChemistries 2097.3.6 ReversiblePEGylation 2097.4 PEGylatedProteinPurification 2107.4.1 RemovalofLow-Molecular-WeightContaminants 2107.4.2 RemovalofFreePEG 2127.4.3 SeparationofPEGylatedandNativeProteinForms 2137.4.4 SeparationofPEGylatedSpecies 2157.5 Conclusions 217

References 218

PartThree AdvancesinProcessDevelopment 223

8 AffinityChromatography:HistoricalandProspectiveOverview 225LauraRowe,GraziellaElKhoury,andChristopherR.Lowe

8.1 HistoryandRoleofAffinityChromatographyintheSeparationSciences 225

8.1.1 Introduction 2258.1.2 EarlyHistory 2268.1.3 BiologicalLigands 2268.1.4 SyntheticandDesignedLigands 2288.1.5 AlternativeLigands 2298.1.6 RoleofAffinityChromatographyintheSeparationSciences 2298.2 OverviewofAffinityChromatography:TheoryandMethods 2308.2.1 BasicChromatographicTheory 2308.2.2 MatrixSelectionandImmobilizationofanAffinityLigand 2328.2.3 OtherConsiderations 2378.3 AffinityLigands 2398.3.1 BiologicalLigands 2398.3.1.1 ImmunoaffinityAdsorbents 2398.3.1.2 BacterialProteins 2428.3.1.3 Lectins 2468.3.1.4 Heparin 2478.3.1.5 Glutathione 2488.3.1.6 AvidinandStreptavidin 2488.3.1.7 VitaminsandHormones 249

X Contents

8.3.1.8 NucleicAcids 2498.3.1.9 AlternativeAffinityMethods 2508.3.2 SyntheticandDesignedLigands 2518.3.2.1 ImmobilizedMetals 2528.3.2.2 HydrophobicLigands 2538.3.2.3 ThiophilicLigands 2538.3.2.4 Histidine 2548.3.2.5 Mixed-ModeAdsorbents 2558.3.2.6 Boronate 2568.3.2.7 BenzhydroxamicAcid 2568.3.2.8 DyeLigands 2578.3.2.9 Biomimetics 2588.4 AffinityLigandsinPractice:BiopharmaceuticalProduction 2698.5 ConclusionsandFuturePerspectives 271

References 272

9 HydroxyapatiteinBioprocessing 283FrankHilbrigandRuthFreitag

9.1 Introduction 2839.2 MaterialsandInteractionMechanisms 2859.2.1 ApatitesforChromatography 2859.2.2 Structure–FunctionRelationship 2899.2.3 RetentionMechanismsinApatiteChromatography 2949.3 SettingupaSeparation 3019.3.1 GeneralConsiderations 3019.3.2 ElutionMode 3059.3.3 DisplacementMode 3099.4 SeparationExamples 3139.4.1 ProteinsinGeneral 3139.4.2 Antibodies 3139.4.3 Polynucleotides 3229.4.4 Others 3239.5 Conclusions 323

References 324

10 MonolithsinBioprocessing 333AlešPodgornik,MilošBarut,MatjažPeterka,andAlešŠtrancar

10.1 Introduction 33310.2 PropertiesofChromatographicMonoliths 33310.3 MonolithicAnalyticalColumnsforProcessAnalyticalTechnology

Applications 33810.3.1 UpstreamApplications 33910.3.2 DownstreamApplications 34010.3.2.1 HPLCAnalysisofIgGProteins 34010.3.2.2 HPLCAnalysisoftheIgMSamples 341

Contents XI

10.3.2.3 HPLCAnion-ExchangeAnalysisofthePEGylatedProteins 34210.3.2.4 Viruses 34410.4 MonolithsforPreparativeChromatography 34810.4.1 ProteinPurification 34910.4.2 PurificationofViruses 35110.4.3 PlasmidDNAPurification 35410.4.4 NegativeChromatography 35710.5 EnzymeReactors 35810.5.1 ProteomeAnalysis 35810.5.2 Biosensors 36010.5.3 BioconversionofTargetMolecules 36010.5.4 StudyofEnzyme-IntrinsicProperties 36210.6 Conclusions 364

References 364

11 MembraneChromatographyforBiopharmaceuticalManufacturing 377OmarM.Wahab

11.1 MembraneAdsorbers–IntroductionandTechnicalSpecifications 377

11.1.1 Introduction 37711.1.2 MembraneAdsorberConstruction 38011.1.3 TypesofAvailableLigands 38211.1.4 UseandScaling-UpwithMembraneAdsorbers 38411.2 ComparingResinsandMembraneAdsorbers 38711.2.1 Flow-ThroughPolishingApplications 38911.2.2 Bind-and-EluteApplications 39011.2.3 EconomicalModelingandCaseStudies 39111.3 MembraneChromatographyApplicationsandCaseStudies 39311.3.1 ValidationofMembranesintoaPurificationProcess 39311.3.2 VirusPurificationandVaccineManufacture 39511.3.3 VirusRemoval 39611.3.4 EndotoxinRemoval 39911.3.5 HCPRemoval 40211.3.6 DNARemoval 40411.3.7 AggregateReduction 40411.4 Conclusions 406

References 407

12 ModelingandExperimentalModelParameterDeterminationwithQualitybyDesignforBioprocesses 409ChristophHellingandJochenStrube

12.1 Introduction 40912.2 QbDFundamentals 41012.3 ProcessModelingandExperimentalModelParameter

Determination 411

XII Contents

12.3.1 Modeling 41312.3.2 ExperimentalModelParameterDetermination 41412.3.2.1 IsothermParameters 41412.3.2.2 FluidDynamics 41612.3.2.3 MassTransferKinetics 41712.4 ProcessRobustnessStudy 42512.4.1 ModelError 42512.4.2 ModelParameterDeterminationError 42612.4.3 VariationofProcessConditions 43112.5 Conclusions 43912.6 Nomenclature 440 Acknowledgments 441

References 442

Volume2

PartFour AnalyticalTechnologies 445

13 BiosensorsintheProcessingandAnalysisofBiopharmaceuticals 447SriramKumaraswamy

13.1 Introduction 44713.2 PrinciplesandCommercialApplicationsofBiosensors 44813.2.1 LabeledversusLabel-FreeBiosensors 44913.2.2 Label-FreeBiosensors 45113.2.2.1 Label-FreeBiosensorsinCommercialUse 45113.2.2.2 IntroductiontoBLI 45313.2.2.3 IntroductiontoSPR 45313.2.2.4 IntroductiontoRWG 45513.2.3 SampleHandlingConsiderations 45513.2.3.1 SampleHandlingbyBLI 45613.2.3.2 SampleHandlingbySPR 45613.2.3.3 SampleHandlingbyRWG 45813.2.4 ComparisonofBiosensorChips 45813.2.4.1 OctetDipandReadBiosensors 45913.2.4.2 BiacoreChips 45913.2.4.3 EpicMicroplates 46213.2.5 ComparisonofThroughput 46213.3 UseofBiosensorsinBiopharmaceuticalProductionand

Processing 46413.3.1 QuantificationofTherapeuticsandOtherMinorImpurities 46413.3.2 PurificationonChromatographyColumnsinDownstreamProcess

Development 46513.3.3 KineticAnalysisforCharacterizationofBiopharmaceuticals 466

Contents XIII

13.3.4 VaccineDesignandEfficacy 46813.4 Conclusions 469

References 470

14 ProteomicsToolkit:ApplicationsinProteinBiologicalProductionandMethodDevelopment 473GlenwynKempandAchimTreumann

14.1 Introduction 47314.1.1 ProblemofAvailability 47414.1.2 WhatIsProteomics? 47414.2 ApplicationsofProteomics 47514.2.1 ProteinIdentificationandCharacterization 47514.2.2 ProteinModifications 47614.2.3 ProteinInteractions 47614.2.4 ProteinQuantitation 47714.3 MythsandMisconceptions–PerceivedDrawbacksofProteomics 47714.3.1 HighSet-UpCost 47714.3.2 Time-Consuming/LowThroughput 47814.3.3 ExpertiseandTraining 47814.3.4 Reproducibility 47914.4 CriticalFactorsforIndustrializationofProteomics 48014.4.1 QualityControl 48014.4.2 RobustnessandReliability 48114.5 CaseStudies 48114.5.1 Two-DimensionalPAGE 48114.5.2 MassSpectrometryasaProcessDevelopmentTool 48214.5.2.1 Matrix-AssistedLaserDesorptionIonizationBiotyping 48314.5.3 QuantitativeProteomics 48414.5.3.1 StableIsotopeLabeling 48414.5.3.2 IsobaricLabeling 48514.6 Conclusions 486

References 487

15 ScienceofProteomics:HistoricalPerspectivesandPossibleRoleinHumanHealthcare 489NawinMishra

15.1 Scienceof“Omics” 48915.2 MajorAdvancesinBiologyThatLedtotheSciencesof“Omics” 48915.3 Mendel’sPrinciplesofInheritance 49015.4 OneGene/OneEnzymeConceptofBeadleandTatum 49015.5 Watson–CrickStructureofDNA 49015.6 DevelopmentofDifferentTechnologiesResponsiblefortheEmergence

ofGenomicsandProteomics 49115.6.1 Genomics-SpecificTechnologies 491

XIV Contents

15.6.2 ProteinSeparation,ProteinSequencing,andTheirThroughputTechnologies 492

15.7 Genomics 49215.8 Proteomics 49315.8.1 StartofProteomics 49615.8.2 DevelopmentofProteomics 49815.8.2.1 Two-DimensionalGelElectrophoresis 49815.8.2.2 MassSpectrometry 49915.8.2.3 X-RayCrystallographyandNuclearMagneticResonance

Spectroscopy 50115.8.3 ProteomicsasaBasisforDifferentiation 50115.9 Interactomics:ComplexityofanOrganismBasedontheInteractionsof

Proteins 50115.10 RelationbetweenDiseases,Genes,andProteins:Diseasome

Concept 50315.11 ProteinsasBiomarkersofHumanDiseases 50315.11.1 ModificationofProteins 50315.12 Metabolomics 50515.13 ProteomicsandDrugDiscovery 50615.14 CurrentandFutureBenefitsofProteomicsin

HumanHealthcare 50615.14.1 UnderstandingComplexDiseasesandPossibilityofPersonalized

Medicine 50615.14.2 BetterDrugsforHumanDiseases 50715.14.3 IdentificationofProteinBiomarkers 50715.14.4 DrugDevelopment 50715.14.5 DiscoveryofNewProteinsasDrugs 50715.14.6 ProteinsLinkedtoBrainDiseases 508

References 508

PartFive QualityControl 511

16 ConsistencyofScale-UpfromBioprocessDevelopmenttoProduction 513StefanJunne,ArneKlingner,DirkItzeck,EvaBrand,andPeterNeubauer

16.1 InhomogeneitiesinIndustrialFed-BatchProcesses 51316.2 EffectsofConditionsinIndustrial-ScaleFed-BatchProcessesonthe

MainCarbonMetabolism 51516.3 EffectsofConditionsinIndustrial-ScaleFed-BatchProcessesonAmino

AcidSynthesis 51816.4 Scale-DownReactorsforImitatingLarge-ScaleFed-BatchProcess

ConditionsattheLaboratoryScale 52016.5 ImprovedTwo-CompartmentReactorSystemtoImitateLarge-Scale

ConditionsattheLaboratoryScale 523

Contents XV

16.6 DescriptionoftheHydrodynamicConditionsinthePFRPartofthePresentedTwo-CompartmentReactor 526

16.7 DescriptionofOxygenTransferinthePFRPartoftheTwo-CompartmentReactor 529

16.8 E. coliFed-BatchCultivationsintheTwo-CompartmentReactorSystem 531

16.9 FuturePerspectivesfortheApplicationofaTwo-CompartmentReactor 537References 538

17 SystematicApproachtoOptimizationandComparabilityofBiopharmaceuticalGlycosylationThroughouttheDrugLifeCycle 545DarylL.Fernandes

17.1 CostsofInconsistent,UnoptimizedDrugGlycosylation 54517.2 Scheme1:TraditionalApproachtoComparabilityofDrug

Glycosylation 54717.2.1 IncomparableGlycosylationDuringScale-UpofMyozyme® 54817.2.2 WhyIncomparableGlycosylationOccurswithTraditionalDrug

Scale-Up 54917.3 Scheme2:ComparabilityofDrugGlycosylationUsingQbDDS 55117.3.1 QbDApproachtoGlycosylationintheA-MAbCaseStudy 55217.4 Scheme3:EnhancedQbDApproachtoComparabilityofDrug

Glycosylation 55417.4.1 InformaticsToolsforEnhancingQbDforGlycoproteinDrugs 55417.4.2 CaseforaPopulationModelforComparabilityofGlycoprotein

Therapeutics 55517.4.3 DomainOntologyModelforDrugRealization 55717.4.4 OntologyMap 55717.4.5 ElementsViewoftheOntologyMap 56017.4.6 BuildingaPopulationComparabilityModelforDrug

Glycosylation 56117.4.6.1 SEBoard 56217.4.6.2 Step1:CategorizetheBiologicalBehaviorsoftheDruginTermsof

SafetyandEfficacy 56317.4.6.3 Step2:DetermineandPrioritizetheGlycosylationCriticalQuality

Attributes 56317.4.6.4 Step3:DevelopaTunedGlycoprofilingSystemtoMeasurethe

GCQAs 57117.4.6.5 Step4:DescribingandOptimizingtheGlycosylationQTPPby

GlycoformActivityModeling 57317.4.6.6 UsingGlycanActivityModelinginGlycosylationOptimizationand

ComparabilityStudies 57717.5 Conclusions 580 Acknowledgments 581

References 581

XVI Contents

18 QualityandRiskManagementinEnsuringtheVirusSafetyofBiopharmaceuticals 585AndyBailey

18.1 Introduction 58518.2 QRMandVirusSafety 58618.2.1 ProductComplexityandRisk 58718.3 PillarsofSafety 59018.3.1 Sourcing–DefiningtheBaselineRisk 59018.3.1.1 Epidemiology–APowerfulToolforReducingRiskforHuman-and

Animal-DerivedComponents 59218.3.1.2 AdditionalMeasuresforControllingAnimal-DerivedMaterials 59618.3.2 Testing–ReducingFurthertheBaselineRisk 59618.3.2.1 In VitroandIn Vivo AdventitiousAgentTests–Advantagesand

Disadvantages 59718.3.2.2 InfectivityTestsforEndogenousRetroviruses 59718.3.2.3 ElectronMicroscopyTestsforRetroviruses 59818.3.2.4 ReverseTranscriptaseAssays 59818.3.2.5 PCRTesting–AdvantagesandDisadvantages 59918.3.3 SourcingandTesting–IsItEnough? 59918.3.4 PathogenClearance–ControllingtheResidualRisk 60018.3.5 ControllingSuppliersofMediaandOtherActivePharmaceutical

Ingredients 60118.4 CommitteeforProprietaryMedicinalProductsGuidelinesfor

InvestigationalMedicinalProducts–RiskManagementinPractice 60218.4.1 UsingGenericDatatoReduceVirusSafetyTesting 60318.4.2 ExperiencewithWell-CharacterizedCellLines 60318.4.3 ReducingVirusValidationRequirementsforIMPs 60418.4.4 PlatformPurificationProcesses 60518.5 DevelopingaRobustRiskMinimizationStrategy–WhatIstheCorrect

Paradigm? 607References 609

19 EnsuringQualityandEfficiencyofBioprocessesbytheTailoredApplicationofProcessAnalyticalTechnologyandQualitybyDesign 613HelmutTrautmann

19.1 Introduction 61319.2 PATandQbDinBioprocessing–EngineeringMeetsBiology 61419.2.1 PATandQbD 61419.2.2 EngineeringMeetsBiology 61619.3 AspectsofBiologicalDemands–SelectedExamples 61719.3.1 BasicPatternsofNutrientMetabolism:GlucoseandGlutamineas

ComplementaryMajorCarbonandEnergySources 61819.3.1.1 GlucoseUtilization 61919.3.1.2 GlutamineMetabolism 62519.3.1.3 GlucoseandGlutamineConcentrationsinBatchCultures 625

Contents XVII

19.3.2 EffectofCultureStatesonGlycosylation 62619.3.2.1 DissolvedOxygenPartialPressureandpH 62719.3.2.2 ConcentrationsofNutrients 62919.3.2.3 ConcentrationsofMetabolicByproducts:LactateandAmmonia 62919.3.2.4 SupplementingSuitablePrecursors 63219.3.2.5 EffectsonSecretedGlycoproteinsintheMedium 63219.3.3 Cell–CellAdhesionandAggregation:InfluenceontheGrowth

BehaviorofCHOCells 63219.3.3.1 Conclusions 63719.4 TechnicalandEngineeringSolutions 63819.4.1 PATandQbDCompliantProcessUnderstandingandProcessControl:

FromDatatoInformationandKnowledge,andItsTransferfromBioprocessDevelopmenttoManufacturing 639

19.4.1.1 AcquisitionofPrimaryData 64019.4.1.2 Gaining/DerivingInformationfromData 64419.4.1.3 ProcessUnderstandingBasedonKnowledge 64619.4.1.4 DemonstrationofProcessUnderstandingandProof-of-Concept 64719.4.1.5 ProcessControl 64819.4.2 ChallengeofSpeedandQualityinBioprocessDevelopment 64919.5 Conclusions 653 Acknowledgments 653

References 654

PartSix ProcessDesignandManagement 657

20 BioprocessDesignandProductionTechnologyfortheFuture 659JochenStrube,FlorianGrote,andReinhardDitz

20.1 Introduction 65920.2 AnalysisofBiomanufacturingTechnologies 66220.2.1 ProcessConceptsinBiomanufacturing 66320.2.2 TotalProcessAnalysis 66620.2.2.1 mAbs 66720.2.3 BatchtoContinuousManufacturing 67220.2.3.1 Discussion 67720.3 AAC:AnythingandChromatography 67920.3.1 Expanded-BedChromatography 67920.3.2 MembraneChromatography 68120.3.3 Liquid–LiquidExtraction 68220.3.4 Crystallization/Precipitation 68420.4 ProcessIntegration 68520.5 ProcessDesignandQbD 68920.6 PackageUnitEngineeringandStandardization 69120.7 DownstreamofDownstreamProcessing 69420.7.1 HumanInsulin 695

XVIII Contents

20.7.2 Antibiotics(Penicillin) 69620.8 Conclusions 699 Acknowledgments 699

References 700

21 IntegratedProcessDesign:CharacterizationofProcessandProductDefinitionofDesignSpaces 707RichardFrancis

21.1 IntroductoryPrinciples 70721.2 OriginalProcessDevelopmentParadigm 70721.3 TheEssentialQbDConcepts 71021.4 Conclusion 715

References 715

22 EvaluatingandVisualizingtheCost-EffectivenessandRobustnessofBiopharmaceuticalManufacturingStrategies 717SuzanneS.Farid

22.1 Introduction 71722.2 ScopeofResearchonDecision-SupportToolsfortheBiotech

Sector 71922.2.1 Challenges 72022.2.2 TypicalStagesofAnalysisandApproaches 72222.3 CapturingProcessRobustnessUnderUncertainty 72322.3.1 Fed-BatchversusPerfusionCultureStrategies 72322.3.2 RobustnessofLegacyPurificationFacilitiestoHigherTiter

Processes 72522.4 ReconcilingMultipleConflictingOutputsUnderUncertainty 72822.4.1 StainlessSteelversusSingle-UseFacilitiesforClinicalTrials 72822.5 SearchingLargeDecisionSpacesEfficiently 73122.5.1 PortfolioManagement:PortfolioSelectionandCapacitySourcing 73122.5.2 ChromatographySizingOptimizationforFutureFacilities 73522.6 IntegratingStochasticSimulationwithMultivariateAnalysis 73622.6.1 PredictingShort-TermFacilityFitUponTechTransfertoLarger

Facilities 73722.7 Conclusions 737 Acknowledgments 739

References 740

PartSeven ChangingFaceofProcessing 743

23 FullPlastics:ConsequentEvolutioninPharmaceuticalBiomanufacturingfromVialtoWarehouse 745RolandWagnerandDethardtMüller

23.1 IncreasedDemand,ReducedVolumes,andMaximumFlexibility–DrivingForcetoPlasticDevices 745

Contents XIX

23.2 Plastic–TheFlexibleAll-RoundReplacer:FromMaterialtoFunction 747

23.3 PollutionwithPlastics:LeachablesandExtractables 75323.4 PlasticsforStorage:VialandBag 75523.4.1 Vial 75523.4.2 Bag 75523.5 PlasticsforCultivation:Flask,Tube,andUnstirredandStirred

Bioreactor 75723.5.1 Flasks 75723.5.2 Tubes 75723.5.3 Bioreactors 75723.6 PlasticsforPurification:ColumnandMembrane 76023.6.1 Column 76023.6.2 Membrane 76123.7 CaseStudy:ComparabilityofPlasticBag-BasedBioreactorsin

CultivationProcesses 76123.8 ConclusionsandProspects 763

References 765

24 BioSMB™Technology:ContinuousCountercurrentChromatographyEnablingaFullyDisposableProcess 769MarcBisschops

24.1 Introduction 76924.1.1 EvolutionofContinuousCountercurrentChromatography 76924.1.2 ContinuousChromatographySystems 77324.1.3 IndustrialApplicationsofContinuousChromatography 77424.1.3.1 FractionationChromatography 77424.1.3.2 ContinuousIon-ExchangeChromatography 77524.2 ContinuousChromatographyinBiopharmaceuticalIndustries 77624.2.1 IndustryDrivers 77624.2.2 PotentialApplicationAreas 77824.2.3 KeyChallenges 77924.2.4 BioSMB™Technology 78024.2.4.1 DisposableFormat 78024.2.4.2 PrepackedColumns 78024.2.4.3 AlternativeChromatographyFormats 78124.3 ProcessDesignPrinciples 78124.3.1 ProcessDesignFundamentals 78124.3.1.1 ThermodynamicEquilibrium 78124.3.1.2 MassTransferKinetics 78224.3.1.3 OtherPhenomena 78324.3.1.4 PerformancePrediction 78324.3.2 ProcessDesignFeatures 78324.3.2.1 FractionationChromatography 78424.3.2.2 CaptureChromatography 785

XX Contents

24.4 CaseStudies 78624.4.1 ProteinAChromatography 78624.4.2 AggregateRemovalUsingHydrophobicInteraction

Chromatography 78724.4.3 VaccinePurificationUsingSize-ExclusionChromatography 78824.5 Conclusions 789

References 790

25 Single-UseTechnology:OpportunitiesinBiopharmaceuticalProcesses 793MaikW.Jornitz,DetlevSzarafinski,andThorstenPeuker

25.1 CurrentSingle-UseTechnologies 79325.1.1 LiquidHoldBags 79425.1.2 Mixing 79525.1.3 ProductandComponentTransfer 79725.1.4 Purification 79825.1.5 Filtration 80025.1.6 SterileConnections 80125.1.7 Filling 80225.2 FutureSingle-UseOperations 80225.2.1 UpstreamOpportunities 80325.2.2 DownstreamOpportunities 80425.2.3 Single-UseProcessEngineering 80425.3 AutomationRequirementsinSingle-UseManufacturing 80625.3.1 DataAcquisition 80825.3.2 MonitoringandControl 80825.3.3 Facility-WideAutomationStructure 80825.4 QualificationandValidationExpectations 80925.4.1 EquipmentQualification 80925.4.2 ProcessValidation 81125.5 OperatorTraining 815

References 815

26 Single-UseBiotechnologiesandModularManufacturingEnvironmentsInviteParadigmShiftsinBioprocessDevelopmentandBiopharmaceuticalManufacturing 817AlfredLuitjens,JohnLewis,andAlainPralong

26.1 Introduction 81726.2 ParadigmShiftatCrucell 81926.2.1 IntroductiontoCrucell 81926.2.2 EvolutionofSingle-UseBiotechnology 82126.2.2.1 PhaseI:Single-UseTechnologyDevelopment–Successwith

Small-ScalePlasticCellCultureUnits 82126.2.2.2 PhaseII:Single-UseBiotechnologiesDevelopment–Scale-Up,

Capsules,andCoupling 824

Contents XXI

26.2.2.3 PhaseIII:Single-UseBiotechnologiesDevelopment–IndustrializationandSimplification 829

26.2.2.4 CrucellManufacturingofmAbswiththePER.C6®CellLine:ACompletelySingle-UseFed-BatchProcess 835

26.2.2.5 MissingElementsandOutlook 83926.2.3 AdaptationofFacilityLayouttoSingle-UseTechnology 84226.2.4 ProcessDevelopmentValueStream 84926.2.5 AssessmentoftheCrucellParadigmShift 85426.3 ConclusionsandGeneralOutlook 856

References 857

Index 859

XXIII

Preface

Over a few decades the advancement of technologies and our understanding and demands has rejuvenated the biotechnology industries in finding biologicals with therapeutic value. Thus, currently over 12 000 large-molecule biotherapeutic prod-ucts are in preclinical discovery or clinical trials around the world today; however, less than one-third of these are in clinical development and very few have found a successful market. As the demand for healthcare products increases around the globe, the need to produce cost-effective therapeutic solutions for the world community has to be met by the biotechnology industries. It is a challenge that the industries have to embrace to face the future and it is clear that the industries have to adapt in order to survive.

The issues at stake are as complex as they are well known. With the current global situation, serious questions of facility financing, and a shift in health-care policy and reimbursement all create a massive burden on strategic plan-ning. The industries realize the need to adapt to face the future in effective manufacturing.

Volume 1 of this book is organized into three parts containing 12 chapters contributed by experienced international scientists. The first two chapters give an overview of strategies for plasmid DNA production from Escherichia coli and advances in protein production technology. Chapters 3–7 give a perspective of the methodologies for protein recovery. An overview of process development is given in Chapters 8–12.

My thanks to all of the authors who have devoted their spare time, and also for their diligence, patience, and goodwill during the production of the volume. They deserve the full credit for the source of the volume.

It is hoped that this volume will be of great value to all those who are involved in the processing and production of bioproducts, and that it will stimulate further progress and advances in this field to meet the ever-increasing demands and challenges.

I should be most grateful for any suggestion that could serve to improve future editions of this volume.

XXIV Preface

Finally, my deep appreciation to Dr. Frank Weinreich of Wiley-VCH for inviting me to edit the volume, and also to Lesley Belfit and her colleagues for their sus-tained support and help.

G. SubramanianMaidenhead, UKJune 2012

XXV

ListofContributors

Lars AumannChromaCon AGTechnoparkstrasse 18005 ZürichSwitzerland

Andy BaileyVirusure GmbHWissenschafts- und TechnologieparkDonau-City-Strasse 11220 WienAustria

Miloš BarutBIA Separations d.o.o.Teslova 301000 LjubljanaSloveniaandThe Center of Excellence for BiosensorsInstrumentation and Process Control – COBIKVelika pot 225250 SolkanSlovenia

Marc BisschopsTarpon Biosystems Inc.Batavia Bioservices B.V.Zernikedreef 92333 CK LeidenThe Netherlands

Eva BrandTechnische Universität BerlinDepartment of BiotechnologyAckerstrasse 71–7613355 BerlinGermany

Yap Pang ChuanUniversity of QueenslandAustralian Institute for Bioengineering and NanotechnologyCorner College and Cooper RoadsBrisbane, Queensland 4072Australia

Vinod B. DamodaranColorado State UniversityDepartment of ChemistryFort Collins, CO 80523USA

Reinhard DitzMerck KGaAPerformance & Life Science Chemicals R&DFrankfuter Strasse 25064293 DarmstadtGermany

XXVI ListofContributors

Graziella El KhouryUniversity of CambridgeInstitute of BiotechnologyDepartment of Chemical Engineering and BiotechnologyTennis Court RoadCambridge CB2 1QTUK

Robert FalconarUniversity of SheffieldChELSI InstituteDepartment of Chemical and Biological EngineeringMappin StreetSheffield S1 3JDUK

Suzanne S. FaridUniversity College LondonAdvanced Centre for Biochemical EngineeringDepartment of Biochemical EngineeringTorrington PlaceLondon WC1E 7JEUK

Conan J. FeeUniversity of CanterburyBiomolecular Interaction CentreDepartment of Chemical and Process EngineeringPrivate Bag 4800Christchurch 8020New Zealand

Daryl L. FernandesLudger LtdCulham Science CentreAbingdon OX14 3EBUK

Richard FrancisFrancis Pharma38 LongmeadowRiverhead, Kent TN13 2QYUK

Ruth FreitagUniversity of BayreuthProcess BiotechnologyUniversitätsstrasse 3095440 BayreuthGermany

Florian GroteClausthal University of TechnologyInstitute for Separation and Process TechnologyLeibnizstrasse 1538678 Clausthal-ZellerfeldGermany

Christoph HellingClausthal University of TechnologyInstitute for Separation and Process TechnologyLeibnizstrasse 1538678 Clausthal-ZellerfeldGermany

Frank HilbrigUniversity of BayreuthProcess BiotechnologyUniversitätsstrasse 3095440 BayreuthGermany

Dirk ItzeckTechnische Universität BerlinDepartment of BiotechnologyAckerstrasse 71–7613355 BerlinGermany

ListofContributors XXVII

Maik W. JornitzSartorius Stedim North America Inc.5 Orville DriveBohemia, NY 11716USA

Stefan JunneTechnische Universität BerlinDepartment of BiotechnologyAckerstrasse 71–7613355 BerlinGermany

Glenwyn KempDream Laboratory LtdMulgrave TerraceGateshead NE8 1AWUK

Arne KlingnerTechnische Universität BraunschweigInstitute of Biochemical EngineeringGaussstrasse 1738106 BraunschweigGermany

Sriram KumaraswamyForteBio Inc.Suite 2011360 Willow RoadMenlo Park, CA 94025USA

John LewisCrucell Holland BVPO Box 20482301 CA LeidenThe Netherlands

Christopher R. LoweUniversity of CambridgeInstitute of BiotechnologyDepartment of Chemical Engineering and BiotechnologyTennis Court RoadCambridge CB2 1QTUK

Linda H.L. LuaUniversity of QueenslandUQ Protein Expression FacilityAIBN BuildingCorner College and Cooper RoadsBrisbane, Queensland 4072Australia

Alfred LuitjensCrucell Holland BVPO Box 20482301 CA LeidenThe Netherlands

Anton P.J. MiddelbergUniversity of QueenslandAustralian Institute for Bioengineering and NanotechnologyCorner College and Cooper RoadsBrisbane, Queensland 4072Australia

Nawin MishraUniversity of South CarolinaDepartment of Biological Sciences715 Sumter StreetColumbia, SC 29208USA

Dethardt MüllerRentschler Biotechnologie GmbHErwin-Rentschler-Strasse 2188471 LaupheimGermany

XXVIII ListofContributors

Thomas Müller-SpäthChromaCon AGTechnoparkstrasse 18005 ZürichSwitzerland

Peter NeubauerTechnische Universität BerlinDepartment of BiotechnologyAckerstrasse 71–7613355 BerlinGermany

Matjaž PeterkaBIA Separations d.o.o.Teslova 301000 LjubljanaSloveniaandThe Center of Excellence for BiosensorsInstrumentation and Process Control – COBIKVelika pot 225250 SolkanSlovenia

Thorsten PeukerSartorius Stedim Biotech GmbHSchwarzenberger Weg 73–7934212 MelsungenGermany

Aleš PodgornikBIA Separations d.o.o.Teslova 301000 LjubljanaSloveniaandThe Center of Excellence for BiosensorsInstrumentation and Process Control – COBIKVelika pot 225250 SolkanSlovenia

Alain PralongCrucell Holland BVPO Box 20482301 CA LeidenThe Netherlands

Kathrin RallaTechnische Universität BerlinDepartment of BiotechnologyAckerstrasse 71–7613355 BerlinGermany

Laura RoweUniversity of CambridgeInstitute of BiotechnologyDepartment of Chemical Engineering and BiotechnologyTennis Court RoadCambridge CB2 1QTUK

Aleš ŠtrancarBIA Separations d.o.o.Teslova 301000 LjubljanaSloveniaandThe Center of Excellence for BiosensorsInstrumentation and Process Control – COBIKVelika pot 225250 SolkanSlovenia

Guido StröhleinChromaCon AGTechnoparkstrasse 18005 ZürichSwitzerland