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Chapter5ProteininactivationsduringchromatographyIintroductionDenaturationAdsorptionisothermIIchromatographytechniquesIIchromatographytechniquesAIon-exchangechromatographyProteinsretentionmodelDisplacementmodechromatographyPurificationofmonoclonalIgMfromhybridomaSilica-basedion-exchangecolumnPoly(asparticacid)-silicaIECExample5.1SeparationofusingHPIECExample5.2SeparationofmRNAsPurificationofX-prolyldipeptidylpeptidaseNewwordsandvocabularies(1)Newwordsandvocabularies(2)PurificationofaminopeptidaseSeparationoflipaseExample5.4separationof(bFGF)andalkalinephosphatasePurificationofbFGFPurificationofβ-glucosidaseAntibioticsMicroheterogeneityMechanisticconsiderationsReversed-phaseHPLCReversed-phaseHPLCExample5.6proteinseparationusingconformationaldifferencesNewwordsandvocabularies(3)DMechanisticConsiderationsTwo-statedenaturationmodelExample5.7kineticsofdenaturationonasurfaceinRP-HPLCKineticsofdenaturationReversibledenaturationmodelSlurry-packedfuse-sillcamicrocolumnsExample:proteindenaturationbyusingSyntheticpeptidepolymerduringRP-HPLCEHydrophobicinteractionchromatographyHICprocessThermalbehaviorofstandardproteinsinHICHICselectivityExample5.8scale-upofHICpurificationofantitumorantibioticSN-07FmechanisticconsiderationsGSubunitexchangechromatographySubunitexchangechromatographyExample5.10immunoaffinitypurificationofantigen(sCDS)Example5.11Purificationofα-amylasebyimmunoaffinitychromatographywithcross-reactiveantibodyHotherchromatographytechniquesCentrifugalpartitionchromatography(CPC)CPCprocessadvantageConclusionsNewwordsandvocabulariesNewwordsandvocabulariesNewwordsandvocabularies(2)QuestionsIIchromatographytechniquesAIon-exchangechromatographyPurificationofmonoclonalIgMfromhybridomaSilica-basedion-exchangecolumnExample5.2SeparationofmRNAsNewwordsandvocabularies(2)Fmechanisticconsiderations
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