用电化学方法测定体外聚集过程中β淀粉样多肽含量（英文）-豆柴文库

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用电化学方法测定体外聚集过程中β淀粉样多肽含量（英文）
Determinationofβ-AmyloidPeptideContentduringInVitroAggregationbyElectrochemicalMethods
Introduction:
Alzheimer'sdisease(AD)isaneurodegenerativedisorderthataffectsmillionsofpeopleworldwide.Itischaracterizedbythepresenceofextracellulardepositsofβ-amyloidpeptide(Aβ)inthebrain.Aβisa42aminoacidpeptidethatisproducedbytheproteolyticcleavageofamyloidprecursorprotein(APP).
TheaggregationofAβintoamyloidfibrilsplaysakeyroleinthepathogenesisofAD.Therefore,thedevelopmentofmethodstomonitorAβaggregationiscriticalforunderstandingthediseasemechanismandfordevelopingpotentialtherapeutics.
Electrochemicalmethodshaveemergedasapowerfultoolformonitoringproteinaggregationbecausetheyaresensitive,selective,andnon-destructive.Inthispaper,wedescribetheuseofelectrochemicalmethodstomeasuretheAβcontentduringinvitroaggregation.
ExperimentalMethods:
1.Materials:
Aβ1-42peptidewaspurchasedfromSigma-Aldrich(St.Louis,MO,USA).Itwasdissolvedin1%NH4OHtoaconcentrationof1mg/mL.Thepeptidesolutionwasthenincubatedat37°Cfor24hourstoallowforself-aggregation.
Humanserumalbumin(HSA)waspurchasedfromSigma-Aldrich.Itwasusedasacontrolproteintodemonstratethespecificityoftheelectrochemicalmethod.
Allotherchemicalswereofanalyticalgradeandpurchasedfromcommercialsources.
2.Electrochemicalmeasurements:
AllelectrochemicalmeasurementswerecarriedoutusingaCHI660Bpotentiostat(CHInstruments,Austin,TX,USA)withaconventionalthree-electrodesystem.Aplatinumwireservedastheauxiliaryelectrode,andaAg/AgClelectrode(3.0MKCl)servedasthereferenceelectrode.Theworkingelectrodewasaglassycarbonelectrode(GCE)withadiameterof3mm.
ThesurfaceoftheGCEwaspolishedusing0.3and0.05μmaluminaslurrysequentially,rinsedwithdeionizedwater,anddriedunderastreamofnitrogengas.
TheAβaggregatesweredepositedontothesurfaceoftheGCEbydrop-casting5μLofthepeptidesolutionontotheelectrodesurface.Theelectrodewasallowedtoair-dryatroomtemperaturefor30minutes.TheAβcontentwasthenmeasuredusingcyclicvoltammetry(CV)anddifferentialpulsevoltammetry(DPV).
ForC