Downstream-processing-of-monoclonal-antibodies—App

JournalofChromatographyB,848(2007)28–39ReviewDownstreamprocessingofmonoclonalantibodies—ApplicationofplatformapproachesAbhinavA.Shuklaa,1,BrianHubbarda,TimTresselb,SamGuhanb,DuncanLowb,∗aProcessDevelopment,AmgenInc.,1201AmgenCourtW.,Seattle,WA98119,UnitedStatesbProcessDevelopment,AmgenInc.,OneAmgenCenterDrive,ThousandOaks,CA91320,UnitedStatesReceived31May2006;accepted8September2006Availableonline13October2006AbstractThispaperpresentsanoverviewoflarge-scaledownstreamprocessingofmonoclonalantibodiesandFcfusionproteins(mAbs).Thistherapeuticmodalityhasbecomeincreasinglyimportantwiththerecentapprovalofseveraldrugsfromthisproductclassforarangeofcriticalillnesses.Takingadvantageofthebiochemicalsimilaritiesinthisproductclass,severaltemplatedpurificationschemeshaveemergedintheliterature.Inourexperience,significantbiochemicaldifferencesandthevarietyofchallengestodownstreampurificationmaketheuseofacompletelygenericdownstreamprocessimpractical.Here,wedescribethekeyelementsofaflexible,genericdownstreamprocessplatformformAbsthatwehaveadoptedatAmgen.Thisplatformconsistsofawell-definedsequenceofunitoperationswithmostoperatingparametersbeingpre-definedandasmallsubsetofparametersrequiringdevelopmenteffort.TheplatformhingesonthesuccessfuluseofProteinAchromatographyasahighlyselectivecapturestepfortheprocess.Keyelementsofeachtypeofunitoperationarediscussedalongwithdatafrom14mAbsthathaveundergoneprocessdevelopment.Aspectsthatcanbereadilytemplatedaswellasthosethatrequirefocuseddevelopmenteffortareidentifiedforeachunitoperation.Abriefdescriptionofprocesscharacterizationandvalidationactivitiesforthesemoleculesisalsoprovided.Finally,futuredirectionsinmAbprocessingaresummarized.©2006ElsevierB.V.Allrightsreserved.Keywords:Monoclonalantibodies;Fcfusionproteins;Cellcultureharvest;ProteinAchromatography;Viralinactivation;Viralfiltration;Ultrafiltration/diafiltration;Processcharacterization;ProcessvalidationContents1.MonoclonalantibodiesandFcfusionproteins...............................................................................292.PurificationofmAbs—literaturereviewandtemplatedpurificationschemes....................................................293.Aplatformapproachtoprocessdevelopment................................................................................314.Aflexible,genericplatformformAbdownstreamprocessing.................................................................314.1.Cellcultureharvestoperations.......................................................................................324.2.ProteinAchromatography..........................................................................................334.3.LowpHviralinactivation...........................................................................................354.4.Polishingchromatographicsteps.....................................................................................354.5.Viralfiltration.....................................................................................................364.6.Ultrafiltration/diafiltration(UF/DF)..................................................................................364.7.Absolutefiltration..................................................................................................375.Processcharacterizationandvalidationactivities............................................................................37Thispaperisapartofaspecialissueentitled“PolyclonalandMonoclonalAntibodyProduction,Purification,ProcessandProductAnalytics”,guesteditedbyA.R.NewcombeandK.Watson.∗Correspondingauthor.Tel.:[phone];fax:[phone].E-mailaddress:[e-mail](D.Low).1Bristol-MyersSquibb,6000ThompsonRoad,EastSyracuse,NY13057,UnitedStates.1570-0232/$–seefrontmatter©2006ElsevierB.V.Allrightsreserved.doi:10.1016/j.jchromb.2006.09.026A.A.Shuklaetal./J.Chromatogr.B848(2007)28–39296.FuturedirectionsinmAbpurification.......................................................................................387.Conclusions.............................................................................................................38References..............................................................................................................381.MonoclonalantibodiesandFcfusionproteinsMonoclonalantibodiesandFcfusionproteinshaveemergedasoneofthemostexcitingtherapeuticmodalitiesinthebiophar-maceuticalindustry.Nineteenmonoclonalantibodiesand3Fcfusionprotein-basedtherapeuticshavebeenapprovedforsaleintheU.S.andtheEuropeanUnion[1]withcombinedannualsalesalreadyexceeding$9billion[2].Severalofthesemoleculesservesignificantunmetmedicalneeds(Table1).Nearlyaquar-terofbiologicsundergoingclinicaltrialsbelongtothisclassofmolecules(PhRMA2004survey,)ensuringthattheimportanceofthisproductclasswillcontinuetoincreaseoverthecomingyears.Someofthecrucialpropertiesofmonoclonalantibodiesforbiologicalapplicationsincludetheirspecificityforinvivodis-easetargetsaswellasthenearinfiniterangeoftargetsforwhichtheycanbegenerated.AlltherapeuticantibodiesareIgGswi