Ifyourpeptideissynthesizedwithaterminalcysteineresidue(i.e.,afreesulfhydrylgroup,-SH),youcanuseanymaleimide-activatedcarrierproteintocreateanimmunogen.One-stepchemicalreactionsareeasywithPierceSulfo-SMCCPre-ActivatedCarrierProteins(Figure1).Ifyourhaptendoesnothaveasulfhydrylgroup,youcanuseEDCchemistry(Figure2)tocoupleamines(lysine)orcarboxylicacids(glutamic,asparticacidor5"-phosphate)tothecarrierprotein.Keyholelimpethemocyanin(KLH),whichisisolatedfromthegiantseamolluscMegathuracrenulata,isoneofthemostpopularcarrierproteinsusedtocreateanimmunogenforinjection.KLHinducesastrongantibodyresponsebecauseofitslargemass(>10⁶M.W.)andbecauseitisanon-mammalianprotein.PiercerecentlyintroducedmaricultureKLH(mcKLH),aproductthatislessdamagingtomarineaquaticenvironmentsbecausethemolluscsareharvestedandmaintainedinsaltwatertanks.Thecharacteristicblue,opalescentcolorofmcKLHishighlysoluble,makingcouplingreactionseasiertocomplete.

Figure1.PeptideCarrierConjugationusingMaleimideActivatedCarriers

Figure2.EDC-MediatedPeptide-CarrierConjugation

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