放大增敏型碳纳米管免疫传感器构筑及胰腺癌肿瘤标志物检测研究

项目名称： 放大增敏型碳纳米管免疫传感器构筑及胰腺癌肿瘤标志物检测研究

项目编号： No.30800247

项目类型： 青年科学基金项目

立项/批准年度： 2009

项目学科： 金属学与金属工艺

项目作者： 陈志春

作者单位： 浙江大学

项目金额： 20万元

中文摘要： 本项目研究了聚电解质以及巴比妥类缓冲液对碳纳米管的分散性能，并研究了基于CNTs的增敏型探针的制备方法，采用层层自组装技术以及伴刀豆球蛋白与辣根过氧化物酶的特异性亲和作用，将HRP-Ab2固定到探针表面，制备了基于碳纳米管的信号放大增敏型探针，通过透射电镜和Zeta电位等对探针进行表征分析。本项目研究了免疫传感器的构筑，构筑了基于半胱氨酸自组装单分子膜的免疫传感器，基于碳纳米管和金纳米簇的免疫传感器，基于(MWNTs-PEI-Au/PB)n多层纳米复合膜的免疫传感器，基于一步法电沉积CS-CNTs-GNPs纳米复合膜的免疫传感器；用扫描电镜、电化学阻抗谱、循环伏安法对免疫传感器进行表征分析。项目进一步研究了免疫传感器的性能，对免疫反应的温度、时间、缓冲液pH等条件进行优化，在最优条件下建立免疫传感器对肿瘤标志物检测的工作曲线，测定传感器的检出限及线性范围，考察免疫传感器的选择性、重复性、再生性及稳定性，并将传感器应用于临床测试。

中文关键词： 免疫传感器；肿瘤标志物；增敏型探针；碳纳米管

英文摘要： This project described the investigation of the dispersity of carbon nanotubes in the presence of polyelectrolyte and barbiturates. Multi-enzyme signal amplification labels, which could maximize the ratio of HRP/Ab2 and amplify the sensitivity, were prepared through layer-by-layer (LBL) assembly and sugar-lectin biospecific interaction between horseradish peroxidase and concanavalin A. The resulting signal amplification labels were characterized by transmission electron microscopy and Zeta potential. Furthermore, four types of immunosensors were fabricated: immunosensor constructed by immobilizing antibody on the CNT-based film decorated with gold nanoclusters, immunosensor based on multilayer films of Prussian blue and multiwalled-carbon nanotube/polyethylenimine/Au (MWNT-PEI-Au) nanocomposite, immunosensor developed by immobilization of antibody on the electrode modified with electrochemically deposited CS-CNTs-GNPs nanocomposite film, immunosensor based on L-cysteine self-assembled monolayer covalently bounded with antibody. The fabrication process and electrochemical behaviors of the immunosensors were characterized by scanning electron microscopy, electrochemical impedance spectroscopy and cyclic voltammetry. The factors influencing the performance of the immunosensors included the buffer pH, the incubation temperature, and the incubation time were investigated. Under optimal conditions, the performance of proposed immunosensors, including linear ranges, detection limit, selectivity, regeneration, reproducibility and stability, were evaluated. The results showed thar the proposed immunosensors could be further applied to clinical detection of cancer biomarkers.

英文关键词： Immunosensor;Tumor marker;Signal amplification labels;Carbon nanotubes