【作者】 殷月芬；

【导师】 黄杉生； 王柯敏；

【作者基本信息】 湖南大学 ， 分析化学， 2003， 硕士

【摘要】 在当今分离和分析科学领域，如何在保证生物活性的前提下，对生物组分进行有效分离和检测，是研究者们普遍关注的热点问题，纳米通道技术为该问题的解决提供了一个新的手段。金纳米通道——作为一种新的固体支撑物，不仅在保证生物活性的前提下，对生物组分进行有效分离和检测，也可以单独作为一种分离工具，又可作为传感器件使用。而且，金纳米通道是多个单通道的阵列，具有单通道的全部性质，同时，多通道又有有效作用面积大的优点，因此开展金纳米通道的研制和应用研究意义重大。 1．以聚碳酸酯滤膜为模板膜，通过化学沉积法，在温和的条件下，制备了膜表面和孔壁镀金的金纳米通道。通过自组装修饰对纳米通道进行修饰；以原子力显微镜对金纳米通道的表面成像进行表征。将金纳米通道安装到自制的流通池上，以金纳米通道为分子迁移和混合物分离载体，建立了一种简单、高效分离的技术。 2．以金纳米通道阵列为迁移载体，开展单分子迁移研究。考察了Al³⁺、Zn²⁺在不同pH值下的迁移，提出了电荷选择机理，并利用该机理合理地解释了迁移结果；考察了色氨酸、桑色素、荧光素、维生素B2在不同的金纳米通道内的迁移，发现分子共平面结构的差异是有机小分子在金纳米通道内的迁移性质的主要因素；首次将有机官能团取代（精细结构）作为参量并结合不同的修饰试剂，考察了氯荧光素、荧光素钠、异硫氰酸荧光素在不同纳米通道内的迁移；首次基于蛋白变性试剂与蛋白质的作用考察生物大分子（牛血清白蛋白和免疫球蛋白）在不同pH值下的迁移，提出了影响蛋白质迁移性质的新因素——蛋白变性因素。率先利用荧光偏振考察了pH值对蛋白质性质的影响，比较了pH值和蛋白变性试剂对蛋白质作用的影响。 3．首次以不同修饰的金纳米通道阵列为分离载体，分离了色氨酸／VB2，在半胱氨酸、异硫氰酸胍修饰的金纳米通道阵列内，色氨酸／核黄素的相对选择分离系数度分别为5.7和8.6，绝对分离度分别为111.5和168.0。控制pH值部分分离了苯胺盐酸盐／罗丹明B。pH值为1.98、5.15、11.98时，相对选择分离系数度分别为1.8、1.2、6.1。基于pH值和蛋白变性试剂的双重作用，分离了牛血清白蛋白/免疫球蛋白，在半肤氨酸、异硫氰酸肌修饰的金纳米通道阵列内，相对分离度分别为31 .6、23.1。该结果表明金纳米通道有类似生物膜的选择透过性，为生物离子开关的仿生模拟提供了一种新的方法。更多还原

【Abstract】 Separation and detection of biological samples without any denaturalization has received much attention. Nanotubule technology, a newly developed but not unimportant method, highlights itself in its transporting and separating activities. It’s been confirmed that solid supports are valuable replacements. Gold nanotubules, as a new kind of support, find their ways in separation tools and sensors with all the characteristics of single nanotubule. Together with other advantages, gold nanotubules, which hold great promise for bioseparation and bioanalysis, can be considered as an important tool of separation and detection of analytes.Gold was deposited on the surfaces and in the pores of the template polycarbonate membranes through a chemical method environment friendly. At room temperature, the gold nanotubule arrays are modified via self-assemble and imaged by atomic force microscope. Based on the fabrication, modification and imaging technology, a new method for the transporting and separating study of molecules is developed.Transporting of single molecules across the gold nanotubule arrays is investigated. The permeating activities of inorganic ions (Al3+ and Zn2+) are studied at different pH levels and explained by electric charge. Then the separation characteristics of tryptophan, morin, fluorescein and vitamin B2 by gold nanotubule modified with different function groups are observed. It is found that the separation of these substances through gold nanotubule is affected by their distinct structures. The transporting behavior of dichlorofluorescein, fluorescein sodium and fluorescein iso-thiocuanate with different substitutional groups is investigated. Then that of BS A and IgG through different modified gold nanotubule arrays at different pH levels is also discussed. The influence of pH levels on protein structure is analyzed using fluorescence anisotropism and a new principle is put forward.Separating of three pairs of moleculies (tryptophan/vitamin B2, hydrochloric aniline/Rhodamine B, and BSA/IgG) is progressed through modified gold nanotubulearrays. For the first pair, the relative separation results are 5.7 and 8.6 respectively and the absolute separation results are 111.5 and 168.0 respectively through cysteine and carbamidine thiocyante modified gold nanotubules. For the second pair, when pH level are1.98, 5.15 and 11.98, 1.8, 1.2 and 6.1 of the relative separation results are obtained. For the third pair, 31.6 and 23.1 are the relative separation results achieved when pH level is 4.5.更多还原