功能化磁性纳米粒子对糖蛋白的分离富集以及唾液酸衍生物合成的研究

发布时间：2018-08-25 13:48

【摘要】：蛋白质的糖基化是蛋白质重要的翻译后修饰之一,对于蛋白质的结构和功能具有非常重要的影响,调控着细胞的许多重要生命过程,是许多临床的生物标志物及治疗的靶标,糖蛋白的研究在生物学和临床医学领域具有非常重要的意义。糖蛋白组学的研究对象一般都是复杂的生物样品体系,低丰度和微观结构的不均一性,使得糖蛋白的分离富集是糖蛋白组学的重点和难点。绪论对糖蛋白的分离富集的几种方法做了简要介绍,重点介绍了分离富集糖蛋白的常用化学反应方法。本论文主要针对糖蛋白的分离和富集的问题开展了研究。利用本研究组制备的具可切割的氧氨基功能基团的磁性纳米粒子对糖蛋白进行了分离富集。磁性纳米粒子上的氧氨基官能团能与唾液酸甘油侧链的邻二醇被高碘酸钠氧化生成的醛基选择性的反应,将糖蛋白富集到磁性纳米粒子上,而可切割的二硫键能在不破坏糖基片段的情况下将糖蛋白释放出来。论文讨论了温度,时间,催化剂,还原剂,p H值,氧化剂的浓度对分离和富集的影响,优化并确立了选择性分离富集糖蛋白的条件。本论文还以唾液酸为起始原料,通过选择性保护与脱保护的方法,在唾液酸的9号位置上引入醛基和叠氮,后续工作将用引入叠氮的唾液酸衍生物对细胞进行代谢标记,再准备与具可切割的炔基功能化的磁性纳米粒子反应,通过点击化学将代谢标记的唾液酸化糖蛋白或糖缀合物进行分离富集。本论文的工作确立了选择性分离和富集糖蛋白的条件,为在复杂生物样品中的应用提供了实验数据。
[Abstract]:Glycosylation of proteins is one of the most important posttranslational modifications of proteins, which has a very important effect on the structure and function of proteins, regulates many important life processes of cells, and is a target of many clinical biomarkers and treatments. The study of glycoprotein is of great significance in the field of biology and clinical medicine. The research object of glycoproteomics is generally complex biological sample system. The low abundance and the heterogeneity of microstructure make the separation and enrichment of glycoprotein become the focus and difficulty of glycoproteomics. In the introduction, several methods for the separation and enrichment of glycoproteins are briefly introduced, with emphasis on the common chemical reaction methods for the separation and enrichment of glycoproteins. This paper focuses on the separation and enrichment of glycoproteins. The glycoprotein was separated and enriched by magnetic nanoparticles with cleavable functional groups of oxygen and amino groups prepared by our team. The O-amino functional groups on the magnetic nanoparticles can react selectively with the aldehyde group formed by the oxidation of o-diol in the side chain of sialic acid glycerol by sodium periodate to enrich the glycoprotein onto the magnetic nanoparticles. The cleavable disulfide bond can release the glycoprotein without destroying the glycosyl fragment. In this paper, the effects of temperature, time, catalyst, reductant pH value and the concentration of oxidant on separation and enrichment were discussed, and the conditions for selective separation and enrichment of glycoprotein were optimized and established. In this paper, sialic acid was used as the starting material, and aldehydes and azide were introduced into sialic acid 9 by selective protection and deprotection. In the following work, sialic acid derivatives introduced azide were used to mark the cell metabolism. Then it is prepared to react with the magnetic nanoparticles with cleavable alkynyl functionalization to separate and enrich the salivary glycosylated glycoproteins or conjugates labeled by metabolism by click-chemistry. In this paper, the conditions for selective separation and enrichment of glycoproteins were established, which provided experimental data for application in complex biological samples.
【学位授予单位】：湖南师范大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：TB383.1;TQ936.2

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