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以特异性识别凝血酶的适配体为分子识别物质,以氨基钌联吡啶衍生物(Ru1)为电化学发光信号物质,基于吡咯/N-(2-羧乙基)吡咯纳米粒子(Ppy-pa NPs)负载适配体和Ru1研制了一种高灵敏检测凝血酶的电化学发光适配体传感器.以N-(2-羧乙基)吡咯和吡咯为单体,采用微乳液聚合方法制备了Ppy-pa NPs.通过EDC/NHS将Ru1与Ppy-pa NPs表面的羧基共价连接制备了Ru1功能化Ppy-pa NPs(Ru1-Ppy-pa NPs).利用核磁共振图谱、傅里叶变换红外光谱和投射电子显微镜图对Ppy-pa NPs和Ru1-Ppy-pa NPs进行了表征.将Ppy-pa NPs Nafion混合物滴涂在石墨电极表面制备成电化学发光化学传感器,可高灵敏检测三丙胺(检出限为3.0×10-8M).通过电化学氧化将对氨基苯磺酸共价键合在石墨电极表面,将5′修饰氨基凝血酶适配体I(TBA-I)共价连接在对氨基苯磺酸修饰的石墨电极表面制备成电化学发光适配体传感器.将5′修饰氨基凝血酶适配体Ⅱ(TBA-Ⅱ)标记在Ru1-Ppy-pa NPs表面制得电化学发光适配体信号探针(Ppy-pa NPs-Ru1-TBA-II).当凝血酶存在时,凝血酶与电极表面的TBA-I特异性结合,再与信号探针结合形成夹心结构,产生很强的电化学发光信号.该传感器具有极低的检出限(3.0×10-16M)和良好的选择性.本工作表明以Ppy-pa NPs为电化学光探针的载体可构建高灵敏度和选择性的电化学发光生物传感器.
An aptamer that specifically recognizes thrombin is a molecular recognition substance, and the amino ruthenium bipyridine derivative (Ru1) is an electrochemiluminescent signal substance based on a pyrrole / N- (2-carboxyethyl) pyrrole nanoparticle (Ppy- pa NPs) loaded aptamers and Ru1, a highly sensitive chemiluminescent aptamer sensor was developed for the detection of thrombin.Microemulsion polymerization was used to prepare N- (2-carboxyethyl) pyrrole and pyrrole monomer Ppy-pa NPs. Ru1-functionalized Ppy-pa NPs (Ru1-Ppy-pa NPs) were prepared by the covalent attachment of Ru1 to the carboxyl groups on the surface of Ppy-pa NPs by EDC / NHS.Using NMR spectroscopy, Fourier transform Infrared spectroscopy and transmission electron microscopy were used to characterize the Ppy-pa NPs and Ru1-Ppy-pa NPs. The Ppy-pa NPs Nafion mixture was applied to the surface of graphite electrode to prepare electrochemical chemiluminescence sensor. (Detection limit 3.0 × 10-8 M) .At the same time, sulfanilic acid was covalently bonded to the graphite electrode by electrochemical oxidation, and the 5 ’-modified amino thrombin aptamer I (TBA-I) was covalently linked Electrochemical chemiluminescence aptamer sensor was prepared on the surface of p-aminobenzene sulfonic acid modified graphite electrode.The 5 ’modified amino condensate (Ppy-pa NPs-Ru1-TBA-II) was prepared on the surface of Ru1-Ppy-pa NPs by TBA-Ⅱ labeling.When thrombin was present, Thrombin specifically binds to the TBA-I on the surface of the electrode, which then binds to the signal probe to form a sandwich structure that generates a strong electrochemiluminescence signal. The sensor has very low detection limit (3.0 × 10-16 M) and good The work shows that the Ppy-pa NPs as the electrochemical probe can be constructed with high sensitivity and selectivity of the electrochemiluminescence biosensor.