WPC} Q~X4ۢ<--7]?<۷1wmhIXs*553+EOhS}eM}\ot[ڔk@][7Hxo<U uJgW~HXDL[Z/@R- URN"f}q1wC6KH6Я-dԋ)7m><@qO⧣A: 20G k7!! VI|㟫(͏ah_1׮JIm C ; &QIP s/5֟!@^ۋ<#OUv+:YIZiƣd>' T#U.>1jD%-F;)U0|%XtnVƒ7waG[lBA#MAg1.}7h3< 7UQ8#U. %nN[ 0L]UNw4 0h EU UBW W 0h UBX X  , m. E 0 e ^ ppU@||| 0U#UBU> Z B`<6X9`(CourierX'  X. Identification of AntibodiesLecture  0 .   #|x<6X9`(CourierXx6X@`7X@($. $      (9 Z 6Times New Roman Regulara1   z)     _9A" ) xdE9W!A X XZXXXXXXXZBXXX.IdentificationofUnexpectedAlloantibodies?T$MLAB2431#C!B# C!ԀXrX #XZXXXr# XXXXZ "0@..  # XX Xʟ##C!X #BC!Ԁ  98  #C!Bx#(@3z$ . !       ,hAZ*Helvetica Regular M z*     _A" ) xdEWxA X XZXXXC!XXZBC!0  0` (#(#0 ` (#` (##C!B#(@3z$ . !       ,hAZ*Helvetica Regular Fy MNSi HCL DTT Sda HTLARgYk Kna McCa JMHjUA, B,Level 1Level 2Level 3Level 4Level 54#,2Quick A.  .0 rjH2 (G Z(Times New Roman >$"Small Circle"0..hp LaserJet 1300 PCL 5e,,,,0,h Z*Helvetica Regular<\  `(&Roman 10cpi12ptH d . !       _XXHHb XXXXU   XXXX8TXXdd8X.IdentificationofUnexpected_Alloantibodies_#XXXX# X & %XX *X,X,` XX*& % %&A.  Introduction  <X,X XX,X,` X<  1.0  Unexpected_alloantibodies_ԀareantibodiesotherthannaturallyoccurringantiAorB. t$$   2.0  Suchantibodiesarefoundinsome0.32%ofthepopulation,dependinguponthegroupofpatientsor ~& donorstudiedandthesensitivityofthetestmethods.V$$ 0  3.0$$ImmunizationtoforeignRBCantigensmayresultfrom:  $$ BX,X, XX,X XB0    a.0$$pregnancy ^ $$ 0    b.0$$transfusion 6 $$ 0    c.0$$followingdeliberateinjectionwith_immunogenic_Ԁmaterialf  $$ 0    d.0$$Insomeinstancestheimmunizingeventisunknown.> $$ EX ,X XX,X, XE0  4.0$$Onceanunexpectedantibodyisdetectedinprenatalorpretransfusiontesting, itsspecificityshouldbe   determinedanditsclinicalsignificanceassessed. r$$ 0  5.0$$ Aclinicallysignificantantibodyisonethat :z"$$ 0    a.0$$Shortenstheanticipatedsurvivaloftransfused_RBCs_Ԁ,$$ 0    b.0$$Hasbeenassociatedwith_HDN_.$$ 0  6.0$$ Theclinicalsignificancevaries. \$$ 0    a.0$$Someantibodiescausedestructionofincompatible_RBCs_Ԁwithinafewhoursorevenminutes.h$$ 0    b.0$$Othersdecreasetheanticipatedsurvivalbyonlyafewdays.@$$ 0    c.0$$Documentedexperiencewithotherexamplesofthesameantibodyspecificitycanbeusedinassessing  therelativeclinicalsignificanceofanantibody.$$ 0  7.0$$Unfortunately,forsomeantibodiestherearenodataanddecisionsmustbebasedonthepremisethatan H antibodyisnotclinicallysignificantunlessactiveinvitroat37Cand/orbythe_IAT_.x  $$ 0  8.0$$Determiningthespecificityofantibodiesencounteredinpretransfusiontestingisimportantinassessing (" theneedtoselectantigennegativebloodfortransfusion. #$$ 0  9.0$$Patientswith clinicallysignificant antibodiesshouldreceivebloodthathasbeenfoundtolackthe !X% correspondingantigenusingpotentantisera."4 &$$ 0  10.0$$In prenatal testing,knowingthespecificityandimmunoglobulinclassofanantibodyhelpspredictthe <$!( likelihoodof_HDN_.%")$$ 0  11.0$$Whileitisnotcrucialtoidentifyunexpectedantibodiesindonorbloods,suchtestingisoftenundertaken &p$+ fortheprocurementofreagentantiseraorteachingsamples.'H%,$$ % ! ,  ,B  .0    GeneralProcedures,یP)&.$$ Ќ    1.0  Anadequatequantityoftestserumand_RBCs_Ԁisessentialtotheresolutionofanyserologicalproblem. +(0 Eitherserumorplasmamaybeused.+)1$$   ,X*2 _0  2.0$$An_EDTA_Ԅ_anticoagulated_Ԁbloodsampleispreferredforstudiesofautologous_RBCs_Ԁtoavoidproblems X associatedwiththeinvitrouptakeofcomplementcomponentsby_RBCs_Ԁthatoccurswhenclottedblood 0 samplesareused.$$        |    X     0  3.0$$MedicalHistory.`$$ 0    a.0$$Itisusefultoknowapatient'sclinicaldiagnosis,numberofpregnancies,transfusionhistoryanddrug h therapy.@$$ 0    b.0$$Arecenttransfusionmaynecessitatetheuseofproceduressuchasredcellseparationtechniquesto    determinethebloodtypeoftheautologousredcells. p $$ 0  4.0$$Itisappropriatetotesttheserumunderinvestigationatalltestphasesatwhichantibodyactivitywas x  initiallydetected.P $$ 0    a.0$$Additionalantibodiesmaybecomeapparentatdifferenttestphases. $$ 0    b.0$$Reactivityofsomeantibodiesmaybeincreasedby extendingtheincubationtime,lowering X temperatures,increasingtheserumtocellratioorbyusingsensitivemethodssuchasenzyme 4 (_ficin_)techniques. d $$ 0  5.0$$Advantagesofusingenzymetechniquesare:$$ 0    a.0$$EnhancesreactivityofRhantibodiesandcomplementbindingexamplesofantiLeaandanti_Jka_.l$$ 0    b.0$$Denaturessomebloodgroupantigens,especially M,N,S,andFy ,soifmultipleantibodiesare t presentandonehasspecificityforMNSorFy,itaidsinidentificationofotherantibodies.N$$   6.0  Theserumunderinvestigationshouldbetestedbythedesiredtechniqueswithapanelofeightormore  groupOreagentRBCsamplesofknownbloodgroupphenotype.~$$ 0    a.0$$Tobefunctional,areagentRBCpanelmustmakeitpossibletoidentifywithconfidencethemost . frequentlyencountered,clinicallysignificant_alloantibodies_.^ $$ 0    b.0$$Adistinctpatternofreactivityshouldbeapparentformostexamplesofsingle_alloantibodies_."$$ 0    c.0$$TheremustbesufficientRBCsamplesthatlack,andsufficientRBCsamplesthatcarry,theantigens  f$ thatindividualsfrequentlymakeantibodiesto.!>%$$   7.0  Itisimportanttoknowhowtheserumunderinvestigationreactswiththeautologous_RBCs_Ԁ(autocontrol) F# ' todeterminewhetheralloantibody,autoantibodyorbotharepresentintheserum.$!($$ C.0  ConsiderationsinInterpretingSerologicalResults%v#*$$   1.0  _Alloantibodies_Ԁofsomebloodgroup_specificities_Ԁ frequently displayconsistentserologiccharacteristics, ~'&%, itisimportanttolookforcharacteristicssuchas:X(&-$$ 0    a.0$$Whataretheeffectsoftemperature,suspendingmediumorenzymesonthereactionwithaparticular 0)&. cellsample.*'/$$ 0    b.0$$IsthereanyvariationinthestrengthofagglutinationobservedamongreactiveRBCsamples.*(0$$ 0    c.0$$Ishemolysispresent.+`)1$$ 0    d.0$$Istheautocontrolpositiveornegative.,8*2$$  @.+4 0  2.0$$Single_alloantibodies_.X$$ 0  0$$a.0$$Itisusuallyeasytorecognizethespecificityofasinglealloantibodythatyieldsclearcutpositiveand  negativereactionswithreagentRBCsamples.$$ 0  0$$b.0$$Whilethetestphaseatwhichaserumreactsissuggestiveofspecificity,itisimportanttoremember 8 thatexceptionalexampleswillbeencountered.h$$ 0  0$$c.0$$Thestrengthofobservedreactionsmayvarydueto dosageaffect ,variationintheamountofantigen   onthecell,deteriorationoftheantigenduringstorageorthepresenceofmultipleantibodies.  $$ 0  0$$d.0$$Althoughaserumdisplaysareactionpatternindicatingasingleantibody, itisimportantto  J  rememberthatadditionalantibodiesmaybepresent. ~ & $$ 0  3.0$$SpecialconsiderationswithRhantibodies.2 $$ 0    a.0$$IfantiEisidentifiedintheserumofatransfusioncandidate,theadditionalpresenceofanticshould   beconsidered.b$$ 0    b.0$$DeterminetheRhphenotypeofthepatient.Evenwhenanticisnotdetectable,itisadvisabletoselect j c,E(R1R1)bloodfortransfusiontoR1R1patientswithantiEsinceanticisa commoncauseof B delayed_HTR_. $$ 0  0$$c.0$$Thereversesituationcauseslessofaproblem.ItanticisidentifiedtheadditionalpresenceofantiE v maynotbedeterminedunlessrareRzR1_RBCs_Ԁareused.Also,almostallcdonorunitswillbeE.N$$   4.0  Phenotypeofautologousredcells.V$$ 0    a.0$$Onceanalloantibodyhasbeenidentifiedinaserumitisnecessarytotestthepatient_RBCs_Ԁforthe  correspondingantigen.$$ 0    b.0$$Whenanalloantibodyispresentintheserum,thecorrespondingantigenwillbeabsentonthe 6 autologous_RBCs_.f $$ K"X ,X  hXX ,X XK0  0$$^^1)0 $$Whenthepatienttypesnegativefortheantigentowhichtheantibodyisdirected,thisprovides " additionalconfirmationofantibodyspecificity.# $ $ 0  0$$^^2)0 $$Ifthepatienttypespositivefortheantigen,thisindicatesmisinterpretationoftheantibodywork !F% up.v" & $ $ 0    c.0$$Antigentyping cannot beperformedonbloodsamplesofrecentlytransfusedindividualsunlessitis &$!( doneona_pretransfusion_Ԁsample or acellseparationtechniqueisperformed(separatingpatient_RBCs_ %") fromtransfused_RBCs_).%#*$$ 0  5.0$$Probability'4%,$$ 0    a.0$$Forconclusiveantibodyidentification,theremustbesufficientreagentRBCsamplestestedthatlack, <)&. andsufficientthatcarry,theantigentowhichanantibodyappearstodisplayspecificity.*'/$$ 0    b.0$$ Theremustbeaminimumofthreecellswhichpossesstheantigenthatreactandthreecellswhich +l)1 lacktheantigenwhichdonotreact .,F*2$$   x- +3 0    c.0$$Itisimportanttorememberthelimitationswhenapatienthasanantibodyagainstamoderatelyhigh X  incidenceantigensuchase.ItisnecessarytotestadditionaleRBCsamplesbeforeconclusively 0 assigningspecificity,andofequalimportance,identifypossible"hidden"or"masked"antibodies.$$ D.0  Multipleantibodies.`$$   1.0  Whenaserumcontainstwoormore_alloantibodies_,itmaybedifficulttointerprettheresultsofserum h studiesusingasinglepanelofreagent_RBCs_.@$$   2.0  Multiple_alloantibodies_Ԁusuallypresentinoneormoreofthefollowingways:   $$ 0    a.0$$Theobservedpatternofreactiveand_nonreactive_Ԁtestsdoesnotfitthatofasingleantibody. J $$ 0    b.0$$ReactionsofvariablestrengthareobservedwiththereactiveRBCsamplesthatcannotbeexplained R  onthebasisofdosage.* $$ 0    c.0$$DifferentRBCsamplesreactatdifferenttestphases. $$ 0    d.0$$Unexpectedreactionsareobtainedwhenattemptsaremadetoconfirmthespecificityofasuspected 2 singleantibody.Forexample,ifaserumsuspectedofcontainingantieisfoundtoreactwith b  additionaleRBCsamples,itispossiblethateitheranotherantibodyispresent, or thesuspected : antibodyisnotreallyantie.InsuchasituationadditioneRBCsamplesmustbetested.$$ E.0  Antibodiestohighincidenceantigens.n$$   1.0  AnalloantibodytoahighincidenceantigenshouldbesuspectedwhenallreagentRBCsamplesare v reactive, buttheautocontrolis_nonreactive _.N$$   2.0  Oncemultipleantibodiesareruledouttheseproblemsareoftenreferredtoan_immunohematology_  referencelaboratory,whichhavethenecessaryrarecellsforperformingsuch_workups_.$$   3.0  Thepatient'ssiblingsareoftenthebestsourceofserologicallycompatiblebloodforpatientswith 0 antibodiestohighincidenceantigens.` $$ F.  Antibodiestolowincidenceantigens. "   1.0  Whenaserumsamplereactsonlywith_RBCs_Ԁfromasingledonorunitthemostlikelypossibilitiesto  h$ considerare:theunitisABOincompatible,thedonorcellshaveapositiveDATorare_polyagglutinable_.!@%$$   2.0  Reactionsbe_tween_ԀaserumandasingledonororreagentRBCsamplemayalsobecausedbyantibodies H# ' tolowincidenceantigens. $!($$   3.0  If_RBCs_Ԁknowntocarrylowincidenceantigensareavailable,theserummaybetestedagainstthem.%x#*$$   4.0  Itisinappropriatetodelaytransfusionwhilesuchstudiesareundertaken,sincefindingcompatibleblood '(%, willnotbeaproblem.X(&-$$ G.0  Anomalousserologicalreactions.*'/$$   1.0  Antibodiestoavarietyofdrugsandadditivescancausepositiveresultsinantibodydetectionand +`)1 identificationtests.,8*2$$   h-+3   2.0  Therearemanytimesthatreactionswillbeobtained,andattemptstoincreasethestrengthofreactivity X  areunsuccessful.0$$       0  a.0$$Aftereveryattempttoidentifythespecificityareunsuccessful,aninterpretationof unableto  determineantibodyspecificityismade.   `   `$$ 0  0$$b.0$$Thepatientmustbetransfusedwithserologically(_crossmatch_)compatibleblood.h$$ ,  ,'LH  .0    SelectedSerologicalProcedures.,'LBLی $$ Ќ  0  1.0$$Whenapatternofreactionsfailstoindicatespecificity,orwhenthepresenceofanantibodyissuspected  p  butcannotbedemonstrated,useofthefollowingproceduresmaybehelpful. H $$ 0  2.0$$ Enzymetechniques areveryusefulinantibodyidentificationstudies.P $$ 0    a.0$$Treatmentof_RBCs_ԀwithproteolyticenzymesenhancesthereactivityofRh,P,I,Lewisand   complementbinding_alloantibodies_Ԁsuchasanti_Jka_. $$ 0    b.0$$AntigensofM,N,SandFyaredepressedordestroyed.4$$ 0    c.0$$EnzymetechniquesshouldbeusedwheneveraweaklyreactiveantibodymaybeanRhantibodyor < whenapatienthasmultipleantibodiesandoneofthemisaFy.$$ 0  3.0$$ Temperaturereduction isusefulfor_alloantibodies_Ԁ(e.g.,antiM,P1)thatreactbetteratcold l temperatures.Specificitymaybecomeapparentatorbelow22C.H$$ 0    a.0$$Anautocontrolisespeciallyimportantfortestsatcoldtemperatures,becausemanyseracontaincold P reactiveautoantibodies.($$ 0    b.0$$AntiIspecificityisconfirmedbytestingthepatientserumwithadult(I+)andcord(i+)cellsat4C.  PositiveswithadultcellsandnegativewithcordcellsconfirmsantiIspecificity.X$$ 0  4.0$$ Increasingtheserumtocellratio increasestheamountofantibodyinthetestsystemwhichmayincrease `  thestrengthofreactivityofantibodiespresentinlowconcentrations.<!$$ 0    a.0$$Increasingtofourdropsisrecommended,butanincreaseof510dropsmaybeindicatedand # extendingincubationtimeupto60minutes,mixingthesolutionperiodically. l$$$ 0  0$$b.0$$Itisimportanttoremovetheserumpriortowashing,as3washeswouldnot h  !  $  @$  @$  t" & adequatelyremovethisvolumeofserum.L# '$$ 0  0$$c.0$$ NOTE :Becautiouswhenattemptingtoincreasetheserumtocellratioin_LISS_Ԁteststhatrequire $") equalvolumesofserumandcells.%#*$$ 0  5.0$$ Increasingincubationtime to3060minutesmayimprovereactivityandhelpclarifytheobservedpattern '0%, ofreactions.d( &-$$   6.0  DecreasingthepHofthereactionmedium to6.5bytheadditionof0.2NHCLtothepatientserum *'/ enhancesthereactivityofcertainantibodies,mostnotable,someexamplesofantiM.*(0$$  P.+4 0  7.0$$Somebloodgroupantibodiesreactpreferentiallyintestsystemsutilizing LowIonicStrengthSalt(_LISS_) X  solutions.4$$ 0    a.0$$_LISS_Ԁreagentaccelerateantibodyuptake(IE,thefirststateofthe_hemagglutination_Ԁreactionthat  involvesassociationofantibodymoleculesto_RBCs_).d$$ 0    b.0$$Avarietyof_LISS_Ԁprocedureshavebeendescribed.l$$ 0  8.0$$ Useof_thiol_Ԁreagents suchas_dithiothreitol_Ԁ(DTT)and2_mercaptoethanol_Ԁ(2ME)cleaveintersubunit   disulfidebondsof IgM molecules.  $$ 0    a.0$$TheIgGmoleculesarerelativelyresistanttosuchcleavage,sotreatmentresultsindestructionofIgM  T  butleavesIgGintactabletoreactinvitro. , $$ Ѐ \   0    b.0$$ TheapplicationsofDTTand2MEin_immunohematology_Ԁinclude: 4 $$ 0  0$$0$$1)0 $$Determiningtheimmunoglobulinclassofanantibody,especiallyiftheantibodyhasthe   potentialofcausing_HDN_.f $ $ 0  0$$0$$2)0 $$DissociatingRBCagglutinatescausedbyIgMantibodies(e.g.,spontaneousagglutinationof n _RBCs_Ԁcausedbypotentcoldreactiveautoantibodies).F $ $ 0  0$$0$$3)0 $$Identifying_specificities_ԀinamixtureofIgMandIgGantibodies,particularlywhenan  agglutinatingIgMantibodymasksthepresenceofIgGantibodies.v $ $ 0  9.0$$_ Prewarmed_ԀTechnique ~&$$ 0    a.0$$Sometimescoldautoagglutininsmaydemonstratehighthermalamplituderesultinginfalsepositive 0 reactionsat37Cand_AHG_.$$ 0    b.0$$Afterconfirmationofthecoldagglutininitisacceptabletoperforma_prewarmed_Ԁantibodyscreenand ` _crossmatch_:8$$ 0  0$$0$$1)0 $$Place_RBCs_Ԁtobetestedinappropriatetesttubesandplacein37Cheatblock.@! $ $ 0  0$$0$$2)0 $$_Prewarm_Ԁtubeofserumto37C." $ $ 0  0$$0$$3)0 $$Add3to4dropsof_prewarmed_Ԁserumto_prewarmed_Ԁcellsandincubate1hour.# $ $ 0  0$$0$$4)0 $$Withoutremovingtubesfromheatblockaddsalinethathasbeen_prewarmed_Ԁto37C. p$ $ $ 0  0$$0$$5)0 $$Immediatelyspinandwash2additionaltimeswiththewarmsaline.!H% $ $ 0  0$$0$$6)0 $$Performthe_AHG_Ԁprocedure.x" & $ $ 0  0$$c.0$$The_prewarmed_Ԁtechniquecanconfirmthatonlyacoldautoagglutininispresentordetectthe ($!( presenceofclinicallysignificantunderlying_alloantibodies_.%")$$ I.0  InhibitionTests&X$+$$   1.0  Somebloodgroupantigensexistinsolubleforminsuchbodyfluidsassaliva,urineorplasma.These `(&- substancesareusefulinantibodyidentificationstudies,eithertoconfirmantibodyspecificitybyinhibition 8)&. ortoneutralizeantibodiesthatmaskthepresenceofconcomitantnon_neutralizable_Ԁantibodies.The *'/ followingsolublebloodgroupsubstancescanbeusedinantibodyidentificationtests.*(0$$  H.+4   2.0  Lewissubstances.X$$ 0    a.0$$LeaandLebsubstancesarepresentinthesalivaofpersonswiththeappropriateLewisphenotype,and  Lewissubstancecanbepreparedfromsaliva.MostbloodbanksusecommerciallypreparedLewis  substance.`$$ 0  0$$b.0$$LewissubstancewillneutralizeLewisantibodiesinapatientspecimen,allowingthedetectionof h underlying,clinicallysignificant_alloantibodies_.@$$ 0  3.0$$P1substance  $$ 0  0$$a.0$$SolubleP1substanceispresentin_hydatid_Ԁcystfluidaswellaspigeoneggs. H $$ 0  0$$b.0$$P1maymaskthepresenceofunderlying_alloantibodies_.TheadditionofP1substancetothepatient's P  serumcausesneutralizationoftheantiP1,allowingthedetectionofunderlying_alloantibodies_.( $$ 0  4.0$$Sda(Sid)substance $$ 0    a.0$$Sdabloodgroupsubstanceispresentinsolubleforminvariousbodyfluids,withthemostabundant 0 sourcebeingurine.`$$ 0  0$$b.0$$Theurineisaddedtothepatientserum,causingneutralizationoftheantiSda.$$ 0  0$$c.0$$Aidsinthedetectionofunderlying_alloantibodies_.h$$ J.0  Titrationp$$   1.0  Thetiterofanantibodyisusuallydeterminedbytestingserialtwofolddilutionsoftheserumagainst   selectedRBCsamples.$$   2.0  Resultsareexpressedasthereciprocalofthehighestserumdilutionthatcausesmacroscopicagglutination.P$$   3.0  Prenatalstudies.X $$ 0    a.0$$Whentheantibodyisofaspecificityknowntocause_HDN_Ԁorwhentheclinicalsignificanceofthe " antibodyisunknown,theresultsoftitrationstudies,outcomeofpreviouspregnanciesareusedto # assesstheneedforamniocentesis. `$$$ 0    b.0$$Risingtitersareindicativeofactiveimmunizationofthemother.h" &$$   4.0  Antibodyidentification$!($$ 0    a.0$$SomeantibodiescauseagglutinationofvirtuallyallreagentRBCsamples,butspecificityisindicated %p#* bydifferencesinthestrengthofreactivitywitheachsampleintitrationstudies.&H$+$$ 0    b.0$$Thetitrationprocedureisnotusedverycommonlyforthispurpose.P(%-$$   5.0  HTLAantibodies.*'/$$ 0    a.0$$HTLAantibodiesreactveryweaklyintheundilutedstatebut,unlikemostweaklyreactiveantibodies +X)1 (e.g.,antiDwithatiterof4),reactatahighdilutions(e.g.,1in2000).,0*2$$ 0    b.0$$SuchantibodiesincludeantiCh,Rg,Cs,Yk,Kna,McCaandJMH.8.+4$$  /,5 0    c.0$$Whenweakreactionsareobservedinthe_IAT_,titrationstudiesmaybeusedtoestablishwhetheror X notthereactionsareduetothepresenceofanHTLAantibody.0$$ K.0  Adsorption$$   1.0  Antibodycanberemovedfromaserumbyadsorptionto_RBCs_Ԁcarryingthecorrespondingantigen.8$$ 0    a.0$$TheantibodyformsacomplexwithRBCmembraneboundantigens.@$$ 0    b.0$$Whentheserumand_RBCs_Ԁareseparated,theantibodyremainsattachedtothe_RBCs_. $$ 0    c.0$$Subsequentelutionoftheboundantibodycanoftengiveadditionalusefulinformation.  $$   2.0  Adsorptiontechniquesareusefulinsituationssuchas: H $$ 0    a.0$$Removingautoantibodyactivitytopermitdetectionofcoexisting_alloantibodies_.P $$ 0  0$$b.0$$Removingunwantedantibodyfromaserumthatcontainsanantibodysuitableforreagentuse. $$ 0    c.0$$Confirmingthepresenceofantigenson_RBCs_Ԁthroughtheirabilitytoremoveaspecificserum X antibody.0$$ 0    d.0$$Confirmingthespecificityofanantibodybyshowingthatitcanbeabsorbedonlyto_RBCs_Ԁofa 8 particularbloodgroupphenotype.$$ 0    e.0$$Separatingmultipleantibodiespresentinasingleserumsample.h$$ L.0  Elutionp$$   1.0  Elutiontechniquesfreeantibodymoleculesfromsensitized_RBCs_Ԁsotherecoveredantibodycanbe   tested .$$   2.0  Avarietyofmethodsareemployedwiththeprimaryobjectivebeingbreakingthebondbetweenthe T antigenandtheantibody.,$$   3.0  Elutiontechniquesareprimarilyusedfor:4!$$ 0    a.0$$Identificationofanantibodycoatingababy's_RBCs_Ԁinthecaseof_HDN_.#$$ 0    b.0$$Identificationofanantibodycausinganacuteordelayedhemolytictransfusionreaction.!<%$$ 0    c.0$$InvestigationofapositiveDAT.D# '$$ 0    d.0$$Concentrationandpurificationofantibodies,thedetectionofweaklyexpressed h  !  $  @$  $") antigensandtheidentificationofmultipleantibodies.%t#*$$ 0  0$$e.0$$Preparationofantibodyfreeintact_RBCs_Ԁforusein_phenotyping_Ԁor_autoabsorption_.|'$%,$$ 0  4.0$$Technicalfactorswhichinfluencethesuccessoftheelutiontechniquesinclude:,)&.$$ 0    a.0$$Incorrecttechnique.*(0$$  <.+4 0    b.0$$IncompletewashingX$$ 0  0$$0$$1)0 $$Ifcellsareincompletelywashedcontaminatingserumantibodywillcausefalsepositive  reactions. $ $ 0  0$$0$$2)0 $$Analiquotofsalinefromthelastwashissavedandtestedinparallel.Positivereactionswith 8 thelastwashinvalidatesthetest.h $ $ 0  0$$c.0$$Bindingofproteinstoglasssurfaces. $$ 0  0$$d.0$$Dissociationofantibodybeforeelution. p $$ 0  0$$e.0$$Instabilityof_eluates_.x $$   ,  ,M  .0    SelectionofBloodforTransfusionAfterAntibodyIdentification,4 ی( $$ Ќ  0  1.0$$Afterantibodyidentificationstudiesarecompletedeterminetheclinicalsignificanceoftheantibody. $$       |   X      `     0  2.0$$Iftheantibodyisclinicallysignificantantigennegativedonorsmustbefoundand_crossmatched_Ԁforthe 0 patient,a_Coomb_s_crossmatch_Ԁmustbedone.`$$ 0  3.0$$Iftheantibodyisnotclinicallysignificantitisnotnecessarytoprovideantigennegativeblood,butthe  donorsmustbecompatiblebythe_Coomb_s_crossmatch_.