基于环糊精和石墨烯衍生物的毛细管电泳涂层柱的研究

发布时间：2018-01-30 11:17

本文关键词： 毛细管电泳蛋白质分离毛细管涂层制备　出处：《青岛大学》2016年硕士论文　论文类型：学位论文

【摘要】：毛细管电泳是一种以高压电场力作为动力,在毛细管内部进行分离的新型的分离技术。毛细管电泳技术是一种高效率,高灵敏度,经济快速的生物大分子分离分析工具,对蛋白质的分离分析更有很大的优势,但是,分析过程中蛋白质会吸附在石英毛细管内壁使分离分析不够准确,为了调整电渗流和抑制涂层对蛋白质的吸附,因此,需要对熔融石英管内壁进行涂层处理,越来越多的毛细管电泳涂层涌现出来,也是目前解决这个难题最主要和常用的方法,常用的毛细管涂层方法主要有化学键合涂层和物理吸附涂层,这两种涂层方式都各自的用途与优点,但是两种方法都有一定的局限性和缺点,其中物理方法制备步骤简洁,但是由于涂层不稳定,分离效果很差,影响分离效果,而化学键合涂层稳定性高,但过程复杂。制备出质量与性能更加优异的毛细管涂层实现蛋白质更好的分离分析效果是该领域研究人员的又一需要突破的重点。本论文采用层层自组装技术(LBL)将大众使用的毒性大、污染性强的硅烷化试剂换做安全环保的水溶性感光高分子重氮树脂,重氮树脂因其自身的结构与特征使其具有独特的光热交联反应,与毛细管涂层新材料反映后可以形成稳定稳固的共价键,涂层的性能更加稳定,对混合蛋白的分离具有很好的效果。本论文主要工作如下:1.使用感光高分子材料重氮树脂(DR)作为偶联剂,通过自组装和共价连接树枝状高分子(CD-Dendrimer)制备毛细管涂层,重氮树脂与树枝状高分子的首先通过离子键固定在毛细管内壁,通过紫外光线照射后,离子键转化为共价键,从而稳定的固定在毛细管内壁,用该种涂层来分离几种混合蛋白质,呈现出良好的分离效果。2.基于层层自组装(LBL)技术,制备出重氮树脂与氧化石墨烯(GO)的毛细管涂层,在毛细管内壁两种分子交替涂层多次,最后在紫外光的照射下,原本重氮树脂与氧化石墨烯之间相互作用的较弱的离子键转化为更加牢固的共价键,从而制备出多层的结构可控的共价键合涂层,用来分离几种蛋白质,分离效果理想,重现性与稳定性也很好。3.选取亲水性物质β-环糊精(β-CD)做为分离蛋白材料,与重氮树脂首先通过离子键在毛细管内表面形成复合膜,起到涂覆毛细管的作用,然后将其放置于紫外线下照射,通过重氮树脂独特的光学反应,是离子键转化为稳定的共价键,毛细管涂层更加稳定,不易脱落,抑制蛋白的吸附作用明显,将制备好的共价键合涂层DR/β-CD毛细管涂层用于分离混合蛋白,效果很好。本论文所研究和制备的毛细管涂层是一种新的偶联剂与新材料的结合,制备出的一种新型毛细管涂层,不仅利用了常用的两种涂层技术的优点,也在一些局限性方面做了很多改善,操作非常简单快速,涂层也很稳定。用以分离混合蛋白来测试其性能,分离结果非常的好,其他测试结果也良好,说明我们所制备的毛细管涂层是稳定有效的,为毛细管的进一步发展提供了更多可能。
[Abstract]:Capillary electrophoresis is a kind of high voltage electric field force as power, new separation technology in the separation capillary inside. Capillary electrophoresis is a kind of high efficiency, high sensitivity, rapid separation and analysis tools of economic biological macromolecules, for protein separation and analysis of advantages, but more substantial, analysis in the process of protein adsorption on quartz the inner wall of the capillary separation analysis is not accurate enough, in order to adjust the electroosmotic flow and adsorption, inhibiting coating on protein therefore need coating of fused quartz tube, capillary tube electrophoresis coating increasingly emerged, is now to solve the problem of the most important and commonly used method, method of capillary coating are mainly used for chemical bonding physical adsorption coating and coating, coating the two way of their uses and advantages, but the two methods have certain limitations and The preparation method of physical weakness, the steps are simple, but because the coating is not stable, the separation effect is poor, affect the separation effect, and chemical bonding coating with high stability, but the process is complex. Preparation of capillary coating more excellent quality and achieve better performance of protein separation effect analysis is the research in this field and a the need to break the key. This paper adopted the self-assembly technology (LBL) will be the mass use of high toxicity and pollution of silanes and strong for safety and environmental protection of water soluble photosensitive polymer diazoresin diazoresin, due to the structure and characteristics of its own. It has unique thermal crosslinking reaction, and can reflect the new material after the capillary coating to form covalent bonds to stabilize, the performance of the coating is more stable, the separation of mixed protein has a very good effect. The main work of this thesis are as follows: 1. using photosensitive polymer Material diazoresin (DR) as a coupling agent by connecting dendrimer self-assembly and covalent (CD-Dendrimer) preparation of capillary coating, diazoresin and dendrimers through ionic bond is fixed on the inner wall of the capillary, through ultraviolet light irradiation, ionic bonds into covalent bonds, and fixed in a stable the inner wall of the capillary, the coating to separate the mixture of proteins, showed a good separation effect of.2. self-assembly (LBL) technique, based on the preparation of diazoresin and oxidation of Shi Moxi (GO) the capillary coating on the inner wall of the capillary, many two molecules alternate coatings, finally under UV irradiation, ionic bond the weak interaction between the original diazoresin and graphene oxide into covalent bonds more firmly, thereby preparing a covalent bond with controllable structure of multilayer composite coating, used for the separation of proteins, The ideal separation effect, reproducibility and stability is also very good.3. selection of hydrophilic substances and beta cyclodextrin (beta -CD) as protein material, resin and diazo through ionic bond in the capillary inner surface to form a composite film, coated capillary action, and then placed in UV irradiation, through optical the reaction of diazo resin is unique, ionic bonds into stable covalent bond, capillary coating is more stable, not easy to fall off, the adsorption inhibition of protein significantly, the covalent bond of prepared composite coating DR/ beta -CD capillary coating for the separation of protein mixture, the effect is very good. This thesis studies and capillary coating system the preparation is a new kind of coupling agent and the new combination of material, a new capillary coating prepared, not only the use of the advantages of the two kinds of coating technology, also some limitations made many improvements, non operation It is simple and fast. The coating is also very stable. It is used to separate the mixed protein to test its performance. The separation result is very good. The other test results are also good. It shows that the capillary coating prepared by us is stable and effective, which provides more possibilities for the further development of capillary.

【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：O658.9

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