Comparison of biosensor platforms in the evaluatio

Comparisonofbiosensorplatformsintheevaluationofhighaffinityantibody-antigenbindingkineticsDanlinYanga,1,AjitSingha,b,1,HelenWua,RachelKroe-Barretta,*aDepartmentofImmuneModulationandBiotherapeuticsDiscovery,BoehringerIngelheimPharmaceuticals,Inc.,Ridgefield,CT06877,USAbTheFuFoundationSchoolofEngineeringandAppliedScience,ColumbiaUniversity,NewYork,USAarticleinfoArticlehistory:Received19April2016Receivedinrevisedform15June2016Accepted24June2016Availableonline27June2016Keywords:BiacoreProteOnOctetMX96Antibody-antigeninteractionsOpticalbiosensorabstractTheacquisitionofreliablekineticparametersforthecharacterizationofbiomolecularinteractionsisanimportantcomponentofthedrugdiscoveryanddevelopmentprocess.Whileseveralbenchmarkstudieshaveexploredthevariabilityofkineticrateconstantsobtainedfrommultiplelaboratoriesandbio-sensors,adirectcomparisonoftheseinstruments'performancehasnotbeenundertaken,andsystematicfactorscontributingtodatavariabilityfromthesesystemshavenotbeendiscussed.Toaddressthesequestions,apaneloftenhigh-affinitymonoclonalantibodieswassimultaneouslyevaluatedfortheirbindingkineticsagainstthesameantigenonfourbiosensorplatforms:GEHealthcare'sBiacoreT100,Bio-Rad'sProteOnXPR36,ForteBio'sOctetRED384,andWasatchMicrofluidics'sIBISMX96.Wecomparedthestrengthsandweaknessesofthesesystemsandfoundthatdespitecertaininherentsys-tematiclimitationsininstrumentation,therankordersofboththeassociationanddissociationrateconstantswerehighlycorrelatedbetweentheseinstruments.Ourresultsalsorevealedatrade-offbe-tweendatareliabilityandsamplethroughput.BiacoreT100,followedbyProteOnXPR36,exhibitedexcellentdataqualityandconsistency,whereasOctetRED384andIBISMX96demonstratedhighflexi-bilityandthroughputwithcompromisesindataaccuracyandreproducibility.Ourresultssupporttheneedfora“fit-for-purpose”approachininstrumentselectionforbiosensorstudies.©2016TheAuthors.PublishedbyElsevierInc.ThisisanopenaccessarticleundertheCCBY-NC-NDlicense().IntroductionThecharacterizationofantibody-antigeninteractionsisessen-tialforthesuccessfuldevelopmentofantibody-basedtherapeutics.Label-freeopticalsurfaceplasmonresonance(SPR)biosensors,thegoldstandardformeasuringthebindingaffinityandkineticsofmolecularinteractions,areusedacrossmultiplestagesofdrugdiscoveryanddevelopment[1e3].SPRbiosensorshavebeenusedtocharacterizeantibody-antigeninteractionsforapproximatelytwodecades[4e6],withapplicationsrangingfromthelow-resolutionaffinityscreeningofantibodysupernatants[7],totherigorous,high-resolutionkineticconstantdeterminationsofpuri-fiedantibodies[8],andtheclassificationofantibodybindingepi-topesviaepitopebinningstudies[9].Theavailabilityofhigh-qualitybindingdataenablestheearlyselectionofcriteria-meetingdrugcandidatesandprovidescrucialinformationforpharmacokinetic/pharmacodynamicmodelinginthedesignofclinicaldosingstrategies[10].Withtherapidlyexpandinguseofmonoclonalantibodies(mAbs)andantibody-likescaffoldsforvarioustherapeuticin-dications[11,12],thedemandfortheefficient,rapidandaccurateidentificationoftherapeuticcandidatesthatmeetspecificabbreviations:mAb,monoclonalantibody;SPR,surfaceplasmonresonance;BLI,Bio-LayerInterferometry;CFM,continuousflowmicrofluidics;PCSK9,proproteinconvertasesubstilisinkexintype9;EDC,1-ethyl-3-(3-dimethylaminopropyl)carbodiimidehydrochloride;NHS,N-hydroxysuccinimide;sulfo-NHS,N-hydrox-ysulfocussinimide;EDTA,ethylenediaminetetraaceticacid;CHO,Chinesehamsterovary;Ni-NTA,nickel-nitrilotriaceticacid;SEC,sizeexclusionchromatography;UPLC,ultra-performanceliquidchromatography;ka,associationrateconstant;kd,dissociationrateconstant;KD,equilibriumdissociationconstant;RU,responseunit;Rmax,maximalbindingresponse;RL,ligandresponse.*Correspondingauthor.DepartmentofImmuneModulationandBiotherapeuticsDiscovery,BoehringerIngelheimPharmaceuticals,Inc.,900RidgeburyRoad,Ridgefield,CT06877-0368,USAE-mailaddress:[e-mail](R.Kroe-Barrett).1Contributedequallytothiswork.ContentslistsavailableatScienceDirectAnalyticalBiochemistryjournalhomepage:://dx.doi.org/10.1016/j.ab.2016.06.0240003-2697/©2016TheAuthors.PublishedbyElsevierInc.ThisisanopenaccessarticleundertheCCBY-NC-NDlicense().AnalyticalBiochemistry508(2016)78e96requirementshasdramaticallyincreased.Duringrecentyears,awidevarietyofinnovativeinstrumentshavebeendevelopedtoaddressthisincreasingdemand,manyofwhichhavedemonstratedsignificantthroughputimprovementsoverthetraditionalBiacoreSPRplatform[13].Theseinstrumentsdifferineitherthedesignofmicrofluidicchannelconfigurationsand/ortheopticalprinciplesinthedetectionofbimolecularinteractions.Examplesincludethe6
6crisscrossmicrofluidicsconfigurationinBio-Rad'sProteOnXPR36,the96-microarrayprintingbytheContinuousFlowMicrospotter(CFM)inWasatchMicrofluidics'sIBISMX96[14,15],andtheBioLayerInterferometry(BLI)opticaldetectiontechniqueinForteBio'sOctetRED384[16]thatiscoupledtoa384-wellhigh-throughputformat.Whilethewell-establishedSPR-based(GE