分子印迹新策略及其在金属离子和蛋白质印迹的应用研究

发布时间：2018-05-24 21:41

本文选题：分子印迹聚合物 + 分子印迹技术　；参考：《曲阜师范大学》2016年硕士论文

【摘要】：分子印迹技术(MIT)模拟自然界中酶与底物之间的相互作用,制备分子印迹聚合物(MIPs)对特定分子进行特异性识别。MIPs以其构效预定性、识别特异性和广泛适用性等三大特点广受关注,各种MIT的策略不断被发展和应用。本文选择两种分子印迹新策略——多模板印迹技术和刺激响应型印迹技术,针对印迹中比较困难的金属离子印迹和蛋白质印迹分别进行了探索,并提出了新思路。主要内容概述如下:1.基于双硫腙螯合作用的多离子印迹材料的制备与应用以Hg(II)、Cd(II)、Ni(II)和Cu(II)为复合模板,双硫腙为离子配体,3-氨丙基三乙氧基硅烷为辅助单体,借助溶胶凝胶技术制备了多离子印迹材料吸附剂(MIIPs)。从双硫腙配位和印迹技术等方面阐释了可能的机理。制备的MIIPs具有较高的吸附容量和快速的吸附动力学,其吸附过程符合郎格缪尔等温模型和拟二级动力学方程。选择性实验和抗干扰实验验证了材料对四种金属离子具有良好的选择性,同时能不受大量共存离子的影响。该MIIPs材料作为固相萃取吸附剂预富集海水中目标离子,其回收范围是94.7% 110.2%,对四种离子的方法检测限在6.0 22.5 ng L-1之间。以多种金属离子作为复合模板,利用一种简单又相对普适的配体,高效的制备印迹吸附剂,能同时识别、富集和去除特定种类的离子,应用前景广阔。2.温敏型分子印迹纳米传感器的制备与应用以藻蓝蛋白为模板,通过自由基聚合,制备一种简单、灵巧的热敏型印迹纳米材料,借助TGA-GSH共修饰后的量子点对藻蓝蛋白进行高效特异识别和灵敏荧光检测,可以控制不同的温度对模板蛋白进行选择性控释。对该传感器进行了表征,对其温度响应、结合能力、选择性、分析性能和应用进行了系统地探究。结果显示,该传感器在0.01-1.8μM的藻蓝蛋白浓度范围有良好的线性关系,方法检测限为10.3 nM。将最终制备的MIPs用于海水样品中,得到加标回收率为92.0%-106.8%,结果满意。本工作提供了一种温和、快速和智能的识别分析水样中痕量蛋白质的有效方法,丰富了蛋白质印迹和刺激响应型分子印迹的相关研究。
[Abstract]:Molecular imprinting (MIT) simulates the interaction between enzyme and substrate in nature, and the preparation of molecularly imprinted polymer (MIPs) for the specific recognition of specific molecules. MIPs have attracted much attention for their predefined structure-activity, recognition specificity and wide applicability. Various MIT strategies are constantly being developed and applied. In this paper, two new strategies of molecular imprinting, multi-template imprinting and stimulation-responsive imprinting, are selected to explore metal ion imprinting and Western imprinting, which are difficult in imprinting, respectively, and a new idea is put forward. The main contents are summarized as follows: 1. Preparation and Application of Multiionic imprinting Materials based on the chelation of Dithizone; the composite templates of Hgtadine, CDI, and Cu), and dithizone as ionic ligands, triethoxy silane, as auxiliary monomers. Polyionic imprinted material adsorbent (MIIPs) was prepared by sol-gel technique. The possible mechanism was explained from the coordination of dithizone and imprinting techniques. The prepared MIIPs has high adsorption capacity and fast adsorption kinetics, and the adsorption process is in accordance with Langmuir isothermal model and pseudo-second-order kinetic equation. The selective experiments and anti-interference experiments show that the materials have good selectivity to the four metal ions and can not be affected by a large number of coexisting ions at the same time. The MIIPs material is used as a solid phase extraction adsorbent for preconcentration of target ions in seawater. The recovery range is 94.7% ~ 110.2, and the detection limit of the four ions is between 6.0 ~ 22.5 ng 路L ~ (-1). Using multiple metal ions as composite templates, a simple and relatively universal ligand was used to prepare imprinted adsorbents, which can simultaneously recognize, enrich and remove specific kinds of ions. Preparation and Application of Temperature-Sensitive Molecular imprinting Nanosensor using phycocyanin as template, a simple and dexterous thermosensitive imprinted nano-material was prepared by free radical polymerization. The highly specific recognition and sensitive fluorescence detection of phycocyanin by quantum dots (QDs) co-modified with TGA-GSH can control the controlled release of template protein at different temperatures. The temperature response, binding ability, selectivity, analytical performance and application of the sensor were systematically investigated. The results showed that the linear range of the sensor was 0.01-1.8 渭 M for phycocyanin, and the detection limit was 10.3 nm. The final MIPs was used in seawater samples and the recoveries were 92.0- 106.8 with satisfactory results. This work provides a mild, rapid and intelligent method for the identification and analysis of trace proteins in water samples, and enriches the studies of Western imprinting and stimulus-responsive molecular imprinting.
【学位授予单位】：曲阜师范大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：O631.3

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