单抗研发及理化表征分析Monoclonal Antibody Development and Ph

本文格式为Word版，下载可任意编辑——单抗研发及理化表征分析MonoclonalAntibodyDevelopmentandPh70mMsaltandcanbeproportionedover30-40%ofthepumprange,wellwithinthecapabilitiesofmodernpumps.Oftenagradientholdfor5minutesattheinitialsaltconcentrationisincluded,justincaseamethodisparticularlysensitivewhenbeingtransferredfromoneequipmenttypetoanother.Insuchcasestheholdtimecanbeadjustedtocompensatefordifferencesinthegradientdelayvolumebetweentheinstruments.

Temperaturesinsidethecolumnaredependentonovendesignandplumbingconfiguration.Havingapre-columnheatexchangerinlineoroutoflinecouldmakeaseveraldegreedifferenceinthetemperatureatwhichthecolumnchemistryoccurs.ThisisparticularlyconcerningforbufferswithwhichthepHcanchangerapidlywithtemperature.Figure14showsacomparisonofcolumncompartmenttemperaturesettingsfortwodifferentinstrumentsfromdifferentmanufacturers.Tomakethecorrelation,thermocoupleswerefittedintoT-piecesinthefluidpathinsidethecolumnoven,butjustpriortothecolumn,andtemperaturesweremeasuredforarangeofcolumncompartmentsetpointsandmobilephaseflowrates.Thesemeasurementswereusedtoestimatethetemperatureofmobilephasegoingthroughthecolumnforeachsetpoint.Byequatingthemeasuredfluidtemperaturesforeachflowrate,thecorrelationofcolumncompartmenttemperatureswereplotted.Itisnotedinthiscorrelationthattherewasalsoasignificanteffectofthemobilephaseflowrateonthecorrelation.

Differentdetectorscansometimesyielddifferencesinbaselineslope.Thiscanoccurwhenmovingfromasingle/doublewavelengthdetectorwithareferencebeamtoaphotodiodearray(PDA)detector.TheselectionofanappropriatereferencewavelengthandbandwidthonthePDAcanovercomedetectorvariance.

Fig.14.ComparisonofcolumncompartmenttemperaturesettingsrequiredtoachievethesamecolumnscompartmenttemperaturefortwodifferentHPLCmodelsatdifferentflowrates.Resultsareshownfora4x250mmDionexProPaccolumn.TheAgilent1100HPLCwasconfiguredusingonlythelefthandsideheatexchanger.TheWaters2695HPLCwasconfiguredwiththesolventpre-heaterin-line.

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11.2Methodvalidation

InnovationsinBiotechnology

Beforeananalyticalmethodcanbeincorporatedintoacharacterizationplatformoraqualitycontrolsystem,itmustfirstbedemonstratedthatthemethodissuitableforitsintendedpurpose.GuidelinesforvalidationofanalyticalmethodshavebeenpublishedintheUnitedStatesPharmacopeia,bytheInternationalConferenceonHarmonization(ICH),USFoodandDrugAdministration(FDA),andinpublishedreviews(Bakshi&Singh,2023).Methodsmustbeevaluatedconsideringregulatoryrequirementsandvalidationprocedures.Inotherwords,the“validatability〞ofthesemethodsmustbeassessedbeforeimplementation.Validationtestsincludeprecision,accuracy,andlinearity.Intermediateprecisionistestedbyusingmultipleinstruments,multipleanalysts,andmultiplecolumnlots.Methodsmustbevalidatedanddocumentedaccordingtoregulatoryrequirementspriortoimplementationintoacontrolsystemforlotreleaseofdrugsubstanceanddrugproduct.Robustnessstudiescanalsobeperformedinconjunctionwithmethodvalidation.Ithasbeenourexperiencethatthemostsignificanteffectsonmethodrobustnessare:mobilephasepHvalue,columntemperature,metalcontaminationandcolumnage.

Asystemsuitabilityrangecanbeobtainedfromrobustnessstudies.Thisrangeisoftenbasedonthestandarddeviationofthemeanforaparticularmeasuredcomponent,suchasmainpeakrelativearea.Thesystemsuitabilityrangeindicatestheprecisionofthemethod.pH-IECmaydemonstrateanimprovementinprecisionoverconventionalsalt-basedIEC(Reaetal.,2023).The6σrangesinFigure15,whichpredictsa99%methodsuccessrate,demonstratetheimprovedprecisionofthepH-gradientIECmethodoverconventionalIEC,whichcanhavea6σrangeofupto8%mainpeakrelativearea(Figure15).

Fig.15.Sixsigmarange(±3SD)formainpeakrelativeareaforsaltgradientIEC(diamonds)andpHgradientIEC(circles)foravarietyofmAbs.

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InnovationsinBiotechnologyEditedbyDr.EddyC.AgboISBN978-953-51-0096-6Hardcover,474pagesPublisherInTechPublishedonline17,February,2023PublishedinprinteditionFebruary,2023InnovationsinBiotechnologyprovidesanauthoritativecrystallizationofsomeoftheevolvingleading-edgebiomedicalresearchtopicsanddevelopmentsinthefieldofbiotechnology.Itisaptlywrittentointegrateemergingbasicresearchtopicswiththeirbiotechnologyapplications.Italsochallengesthereadertoappreciatetheroleofbiotechnologyinsociety,addressingclearquestionsrelatingtobiotechpolicyandethicsinthecontextoftheresearchadvances.Inaneraofinterdisciplinarycollaboration,thebookservesanexcellentindepthtextforabroadrangeofreadersrangingfromsocialscientiststostudents,researchersandpolicymakers.Everytopicweavesbacktothesamebottomline:howdoesthisdiscoveryimpactsocietyinapositiveway?HowtoreferenceInordertocorrectlyreferencethisscholarlywork,feelfreetocopyandpastethefollowing:JenniferC.Re