新型单克隆抗体荧光探针染料的合成及免疫荧光组织化学应用-化学(无机化学)专业论文.docx

6．分别用新型单克隆抗体荧光探针染料TET2477acid及聊4457acid对抗生蛋白链菌素单克隆抗体(免疫球蛋白G，kG)进行标记，制备了单克隆抗体荧光探针。经过测定两种荧光探针染料对抗体的取代度均在2～4之间，标记有效，取代度适中。用该单克隆荧光探针进行免疫反应，利用共聚焦荧光显微成像技术得到了牛肺部动脉内皮细胞Q一微管蛋白及牛肺部内皮细胞肌动蛋白标记细胞图像。对u20S(人体骨肉瘤上皮细胞)，Hela(人宫颈癌上皮细胞)用直接法进行标记，得到了标记细胞图像。免疫荧光组织化学应用表明新型单克隆抗体荧光探针染料具有良好的荧光性能和生物特异性，是一种性能良好的单克隆抗体荧光染料。关键词：5(6)．氯代荧光素，荧光探针染料，合成，单克隆抗体，免疫荧光组织化学ABSTRACTF1uorescentprobeshavebeenwidelyusedinmolecularbiology，血crobiology，biochernistryanal”icalche面st彤cheIllicalengineeringandmedicineengineering．Atthes锄etiIlle，tllerequestfortllepropertiesoffluorcscentprobesisbecoIIlingmoreaIldmoreri百d．Therefore，thes)rIlttlesis肌d印plicationresearchofnuorescentprobedyesbecomeahotpointiniI屹rdisciplinaryoflifescienceandche血stry．RuorescienderiVativesareprizedfortheir10ngexcitationwavelengm，lli曲extinctioncoemcient，llighnuorescencequaIltumyieldinaqueoussolutionaIldtheabilityofmeirfluorescencech麟忙teristicstobetailoredforapaIticulaurapplication．Asaresult，tlleyareusedasnuorescentprobedyeswidely．hltllistllesis，asedesofnovelmonoclonalaIltibodynuorescentprobedyesbasedonnuorescicnweredesi印edalldsynmesized．TowofthedyeswereinVestigatedfortlleirapplicationinimmunonuorescencellistocheIIlistry．(1)Thesynthesisconditionsof2，5一dichloro-p—xylenewerestudiedandoptinlized．AnewⅡlethodofsyrlthesisof2，5一dichlOro—p—xyleneunderIIlicrowave wausstudied．1ksynmesisconditionsof3，6一dichloro—trirnelliticanhy嘶dewerestudiedaIIdoptimized．(2)TwonoVelcompounds(4，7，4’，5’-tetraChloro一6一carboxynuoresceinand4，7，4’，5’一tetrac量lloro一5一carboxynuorescei芏1)weredesigIledaIldsynmesized．111estructuresoftheisoIIlerswerei(1entifiedbyⅡ己andMS．Thesynthesisconditionsof5(6)一carboxylc11loronuoresceinderiVati、伧swerestudiedandoptimized．5(and6)一carboxylchlorofluoresceinisonlersweres印aratedsuccessfully．AUofmeisomersweredetenIlinedby1HNMR，瓜，elementalanalysis andMS．(3)4，7，2’，7’-tetrachloro·5(and6)一carboxylnuoresceinisoIIlersweresynthesizedands印aratedsuccessfully．Thec巧stalstmctureof4，7，2’，7’一tetrachloro一5一carboxynuoresceinNHSesterwasdete彻jnedbyX—raydif!fj协ctionfirsny．(4)Fou