基于适配体荧光传感器检测IgE的方法研究-营养与食品卫生专业论文.docx

ABSTRACTImmunogIobulinE(IgE)，asasecreto叮immunoglobulin，playsakeyroleinh啪aIlalle唱icresponses．Nomlally，IgEispresentinsmall锄outsinhumallbloodsemm，ho、Ⅳever，IgElevelsincreaseinpatients谢thalIe玛ic嬲tIl】na，atopicde咖atitisaIldotherimmunede矗ciency-relateddjseaSes．Therefore，it’simponaIltfordiIlicaldiagnosticstodevelopeas呻le，r印id锄dsenSitiveme山odfort11edetectionofIgE．C伽曲illingnuo心Scencespec嘶scopyanda肚姐erposses咖glliglls唰丘c时，t11isaniclee嘲bliShedanuoreScent印t蠲enSorb嬲edontlleprincipleofnIIorescencereSonallce锄erg)，仃{msfe“FI己ET)todetectimmunodobulinE(IgE)．Inthisanicle，CyaIlinedyes—Cy5锄dhydmph订icc曲onnanomaterials—grapheneoxide(GO)werechosenaSthenuoreScentenergydonorandreceptorrespectiVeIy．Theanti—IgEapt锄erwhosetagged埘mCy5collldbe删ilyabso毗donmesurf砬eofGOvia兀-兀conjugateaccumul撕one仃ect．ResonaIlceene珏搿tmsferbe铆eenCy5andG0，couldquenchtllenuorescenceofCy5．Withtheta唱etproteinIgEadded，itcompetedwithGOforbindingto印t锄er．111llS，也enuore：scemsi印alsofCy5re咖red．Intheexperimentalprocess，wefirstlyuSedtheThree—Dimension越ExcitationEmissionmatrixFIuorescenceSpectrophotometer(3D—EMM)toanalyzethepropeniesofCy5tagged印tamer，using633nma11d675nmastheexcitationandemissionWaVelengtll，respectiVely．ThekindofquencherS，incubationtemperatureaIldincubationtimewereoptimized．Theresultssho、vedthattheh追hestnuoreScencesignaJresponSewaSobtainedusing00asthequencheraRerincubationin1xPBS(pH7．4，CM92+1mmol／L)at37℃for30min．Undertheoptimizedconditions，Jinearrangeof0．05—1O姗oi／LwaSobtained，wit}ladetectionlimitof1pmol／L．IgGandHASwereselectedforthespecificityValidationofthepresentmethod．ThereSultindicatedthat，bothshowedlownuorescenceresponse，whichindicatesahi曲selectiVityandspecificityofthismethod．FinaUy，weappl．edtheestablishedmethodtodetecttheIgEinahumanbloods帅ple，andusedtheclassicalELlSAmethodtoveri匆thetestresuIt，whichexhibitedagoodconsistency．Weconcludethatthepresentmethodise伍cient，rapid，sensitiVeandhasgreatpotentialinactIJalclinicalapplications．KeyWords：Apt锄ers，IgE，Fluorescentsensor，G0，DetectiOn万方数据目录1前言．．11．1核酸适配体．11．1．1核酸适配体的筛选21．1