基于HRP和纳米材料的凝血酶电化学适体传感器的研究-分析化学专业论文.docxVIP

2．制备了壳聚糖(cs)．Au纳米复合物，以高效吸附固定适体及酶标HRP，制备Tcs．Au．HRP／Apt生物纳米复合物，并基于该复合物 2．制备了壳聚糖(cs)．Au纳米复合物，以高效吸附固定适体及 酶标HRP，制备Tcs．Au．HRP／Apt生物纳米复合物，并基于该复合物 构建了三明治型凝血酶电化学适体传感器。该法优势在于能在该生物 纳米复合物表面高效固定HRP酶标，有效地提高了酶标的负载量。以 对苯二酚为媒介体，实现了超敏电化学信号的输出，该适体传感器可 检测低至5．5 fM的凝血酶。 3．基于去铁蛋白纳米粒子、HRP的催化性能和磁性纳米粒子的 高富集特性，构建了一种三明治型安培适体生物传感器用于高敏检测 凝血酶。制备金包磁性纳米粒子修饰的适体I(Aptl一AuMNPs)、 适体II和HRP修饰的去铁蛋白纳米粒子(Apt2一HRP．Apo)及凝血酶 的三明治型复合物，并将之磁性分离富集至丝网印刷碳电极表面进行 电化学检测。该方法制备的传感器检测凝血酶的线性范围为0．8．100 pM，检测限可以达No．17 pM(S／N=3)。该方法操作简单，灵敏度 高，特异性强，在蛋白质的检测和疾病诊断应用中具有一定的潜力。 关键词：电化学适体传感器；凝血酶；金纳米粒子；去铁蛋白；壳聚糖； 酶放大；差分脉冲伏安法 II ABSTRACTThrombin ABSTRACT Thrombin is the extracellulal serine protease involved in the coagulation cascade，and it is thought to regulate many processes in inflammation and tissue repair at the vessel wall．In addition，Thrombin also has hormone-like properties，and it is involved in thrombosis and platelet activation．Therefore，thrombin plays a central role in a number of cardiovascular diseases．It is significant to analyze and examine thrombin in the field of medicine．Among various detecting methods， electrochemical biosensor technology has unique advantages such as high sensitivity，time／cost—effectiveness， and portability of measuring equipments and on line monitoring．With the high selectivity and affinity of aptamers as well as the amplification effect of Nano materials and enzymes，this thesis has developed a series of novel electrochemical aptasensors for hi曲ly sensitive detection．The main contents al e summarized as follows： 1．An ultrasensitive and highly specific electrochemical aptasensor for thrombin based on amplification of aptamer-gold nanoparticles -horseradish peroxidase(aptamer-AuNPs-HRP)conjugates has been developed successful．In this electrochemical protocol，aptamer l(Apt 1) was immobilized on core／shell Fe304／Au magnetic nanoparticles (AuMNPs)and served as capture probe．Aptamer2(Apt2)was dual llI labeled labeled with AuNPs and HRP and used as detection probe．In the presence of thrombin