DNA固定在纳米颗粒表面的方法.ppt

磷酸化的DNABiotin DNA炔基修饰的DNA巯基修饰的DNA氨基修饰的DNA 金银纳米颗粒碳纳米管 石墨烯二氧化硅纳米颗粒量子点上转换纳米颗粒 磷酸化的DNA与TiO2 堆积 如单链DNA与石墨烯 碳纳米管静电吸附 1 Biotin DNA 2 炔基修饰的DNA及其他生物分子 3 NH2 DNA 氨基羧基 氨基羧基氨基氨基 1 EDC和NHS Forbestresultsintwo stepreactions performthefirstreactioninMESbuffer orothernon amine non carboxylatebuffer atpH5 6 thenraisethepHto7 2 7 5withphosphatebuffer orothernon aminebuffer immediatelybeforereactiontotheamine containingmolecule 氨基羧基 DCC N N Dicyclohexylcarbodiimide 氨基 氨基 1 BS PEG nPEGCrosslinkers AmineReactive 2 TSAT Tris succinimidylaminotriacetate Water Water NHSesters pH7 9 3 DSS disuccinimidylsuberate 4 BSOCOES bis 2 succinimidyloxycarbonyloxy ethyl sulfone 5 DSG disuccinimidylglutarate 6 DTSSP 3 3 Dithiobis sulfosuccinimidylpropionate Water Water Water DMSOorDMF 8 BS Bis sulfosuccinimidyl suberate 9 EGS ethyleneglycolbis succinimidylsuccinate 10 Sulfo EGS ethyleneglycolbis sulfosuccinimidylsuccinate 7 DST disuccinimidyltartrate Water Water DMSOorDMF DMSOorDMF 4 巯基化的DNA 1Au SAg S2巯基氨基3巯基羧基4巯基羟基5巯基巯基 巯基 氨基 1 AMAS N maleimidoacetoxy succinimideester NHSesters pH7 9 Maleimides pH6 5 7 5 Usually7 2 7 5 Twostepsreaction DMSOorDMF 2 BMPS N maleimidopropyloxy succinimideester DMSOorDMF 3 EMCS N maleimidocaproyloxy succinimideester DMSOorDMF 4 Sulfo EMCS N maleimidocaproyloxy sulfosuccinimideester Water 6 Sulfo GMBS N maleimidobutyryloxy sulfosuccinimideester 5 GMBS N maleimidobutyryloxy succinimideester DMSOorDMF Water 7 BMPA N maleimidopropionicacid EDC 0 1molMESpH4 7Maleimidesreactwithsulfhydryls SH atpH6 5 7 5 8 LC SMCC Succinimidyl 4 N maleimidomethyl cyclohexane 1 carboxy 6 amidocaproate DMSOorDMF 10 Sulfo SMPB sulfosuccinimidyl4 p maleimidophenyl butyrate 9 SMPB succinimidyl4 p maleimidophenyl butyrate DMSOorDMF Water 11 Sulfo LC SPDP LC SPDP SPDP 巯基 羧基 巯基 羟基 异氰酸脂在水溶液中很容易水解 所以最好用有机溶剂如DMSO DMF等 如在水相中 则要使用pH8 5的碱性缓冲溶液 避免使用含羟基的缓冲液如tris缓冲液 且要注意含羟基的分子不能有氨基和巯基 因为pH 7 5顺丁烯二酰亚胺失去对巯基的特异性 会同时与氨基和巯基发生反应 巯基 巯基 pH6 5 7 5 表征 1 定性 红外表征 如交联剂的特征峰 测颗粒的在260纳米处的紫外吸收 跑电泳 电位 粒径变化 2 定量 测颗粒或上清的紫外吸收 荧光 热重力分析 谢谢大家