【作者】 何晓晓；

【导师】 王柯敏；

【作者基本信息】 湖南大学 ， 分析化学， 2003， 博士

【摘要】 本论文瞄准生物纳米技术发展的前沿方向，以核壳纳米颗粒制备、核壳纳米颗粒性能分析、核壳纳米颗粒应用为主线，开展了四方面的研究工作：第一，开展了核壳纳米颗粒的制备方法以及机理的研究，取得了核壳纳米颗粒制备理论和技术的突破。证明了基于硅烷化试剂水解形成的二氧化硅外壳与微乳液方法形成的内核实现稳定的核壳结构结合，关键是由壳材料与内核材料的电性决定的。在核壳纳米颗粒形成机理理论的指导下，发展了二氧化硅为外壳的核壳纳米颗粒制备技术以及氨基化硅壳类纳米颗粒同步制备技术，制备了一系列具有自主知识产权的核壳结构纳米颗粒，并为核壳纳米颗粒的放大制备提供了依据。第二，从体内和体外两个角度系统地研究了核壳纳米颗粒(主要包括二氧化硅纳米颗粒(SiNP)、二氧化硅壳荧光纳米颗粒(FSiNP)以及二氧化硅壳磁性纳米颗粒(MSiNP))的生物亲和性，证明了该类纳米颗粒有很好的生物亲和性。第三，采用透射电子显微镜、原子力显微镜、Zeta电位分析仪、交流梯度磁场计等仪器对该类纳米颗粒的物理化学性质进行了表征。第四，结合生物技术与纳米技术，建立了基于核壳生物纳米颗粒的生物分析新方法，开展了纳米尺度上的原位、活体、实时生物医学分析：(1)开展了纳米尺度上生物与化学信息的荧光传感，一是建立了基于核壳生物荧光纳米颗粒的新型荧光标记方法，应用这一新型荧光标记方法成功地实现了SmIgG~+B淋巴细胞的识别以及系统性红斑狼疮疾病的免疫学检测；二是基于荧光纳米颗粒构建了纳米pH传感器，测量了单个巨噬细胞内的pH变化；(2)构建了基于硅壳磁性纳米颗粒的超顺磁性纳米捕获器，分离了葡聚糖、DNA等物质；(3)发展了一种基于氨基化硅壳类纳米颗粒的新型DNA富集技术，并基于该技术发展了一类基于氨基化硅壳类纳米颗粒的具DNaseI酶切保护性的新型非病毒型基因载体以及基于氨基化硅壳类纳米颗粒的DNA快速抽提纯化方法。苷酸的特异性结合，成功地实现了对互补单链核苷酸片段的高效快速分离和富集，且不改变分离对象的特性。更多还原

【Abstract】 According to the advanced directions of the bionanotechnology, four parts of investigations has been performed in this thesis. In part one, the synthesis methods and mechanism of core-shell nanoparticles have been investigated. Great breakthrough in the synthesis theory and techniques of core-shell nanoparticles has been obtained. The results have testified that the charges of the shell materials and core materials will dominate whether the stable core-shell structure forms or not, by employing the silica shell formed through the hydrolysis of silane agents and the core formed by microemulsion. Under the instruction of the synthesis mechanism theory of core-shell nanoparticles, a series of making-technique of core-shell nanoparticles have been developed. A series of core-shell nanoparticles with own intellectual property have been prepared, which provides theory for the magnified synthesis of core-shell nanoparticles. In part two, the biocompatibility of the core-shell nanoparticles, including silica nanoparticles, fluorescent silica nanoparticles and silica coated magnetic nanoparticles, have’been investigated from the angle of in vivo and ex vivo. In part three, the physical and chemical properties of core-shell nanoparticles have been characterized by using transmission electronic microscopy, atomic force microscopy, zetasizer and alternating grade magnetometer. In part four, new biological analytical methods based on core-shell bio-nanoparticles have been established and the analysis of biological medicine in suit, in vivo, in real time have been developed under the combination of nanotechnology and biotechnology. (1) Biological and chemical fluorescent sensing at nano-scale have been developed. One is the novel fluorescent label method based on core-shell fluorescent nanoparticles. We have recognized the SmlgG+B lymphocyte and detected the Systematical Erythema Lupus by using of this novel label. The other is a developed nano-pH sensor based on the fluorescent nanoparticles. The pH changes in single murine macrophages have been measured by using the nano-sensor. (2) Super paramagnetic nano-captures based on silica coated magnetic nanoparticles have been developed. These captures were successfully used to separate substances like glucan and DNA. (3) A novel DNA enriching technique based on amino-modified silica nanoparticles have been established. A novel non-viral genecarrier with protective ability from DNasel and a rapid DNA extraction and purification method have been developed based on the novel DNA enrichment technique.更多还原