Generic Method .pdf

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1、FeasibilityofDevelopingaGenericcIEFMethodfortheDeterminationofChargeHeterogeneityofAllMabsJiaqiWu27CoronetRoad,Toronto,Ontario,CANADAM8Z2L8Introduction©MethodstodeterminechargeheterogeneityofMabs–SlabgelIEF»Timeconsuming,laborintensiveandsemi-quantitative–Conv

2、entioncapillaryIEF(cIEFperformedongeneralCE)»Longmethoddevelopmenttime(months),poorresolutionandreproducibility–Ionexchangechromatography»Longmethoddevelopmenttime(months)»variationofresolutionfromcolumntocolumn©Whole-columndetectioncIEF–Shortmethoddevelopment

3、time(hoursforMabs),–Reportofgenericmethodvalidatedfor20Mabs**XiaopingHe,et.Al.,CEPharm2006Whole-ColumnDetectioncIEF-cIEFwithoutMobilization–iCE280Analyzer+-H+-InletOHIEFColumnOutletSemi-permeableSampleZonestubeWhole-columndetectoriCE280AnalyzerFeatures©Mobiliz

4、ationprocessiseliminated–Onlyneedtooptimizefocusingprocess©Whole-columndetectorconstantlymonitorsIEFprocessinentirecolumn©Shortanalysistime(upto6runs/hour)©Acceleratedmethoddevelopment(hoursforMabs)©Reproduciblepeakpatternandhighresolution©WidepIrange(pH2-11)I

5、EFMethodDevelopmentUsingiCE280Analyzer–FlowChartInitialcondition:0.1mg/ml4%pH3-10Pharmalytes,6minAdjustsampleconcentrationIfAbs<0.05or>0.4YesNo1.Adjustfocusingtime2.TryadditivesPeakpatternreproducible?NoYesSelecttwopImarkersanddeterminesamplepI1.Narrowrangeamp

6、holytesormixtureofcarrierNeedhigherampholytesYesresolution?NoResultsIEFMethodDevelopmentforMabsUsingiCE280Analyzer©EasytoachievereproduciblepeakpatternsformostMabs–Noadditivesareneededfor>90%ofMabs–Focusingtimeisoftenoptimizedinasinglerun©FormostMabs,onlythree

7、conditionsneedtobeoptimized–Sampleconcentration–SelectionofnarrowpHrangecarrierampholytestoachievehighresolution–SelectionofpImarkersOptimizationofThreeConditionsinIEFMethodDevelopmentforMabs©Sampleconcentration–Sampleconcentrationshouldbehighenoughtodetectmin

8、orspecies,butnottosaturatethedetector–Majorpeakheightreaches0.5Ab©SelectionofnarrowpHrangecarrierampholytes(CA)–NarrowpHrangeCAsarenecessarytoachievesatisfactoryresolution–TheCAssh