用于检测糖蛋白的荧光印迹纳米材料的制备与应用研究

发布时间：2018-07-03 13:06

本文选题：糖蛋白 + 硼酸　；参考：《山东师范大学》2017年硕士论文

【摘要】：糖蛋白是生物体内一种重要生物大分子,是蛋白质糖基化修饰后的产物,在生物体内发挥重要作用。研究表明,糖蛋白在生物体内表达量的高低与多种疾病相关:如转铁蛋白受体这种膜结合型糖蛋白,它的作用是与转铁蛋白结合,将铁转运到细胞内,供细胞正常的生命活动,当生物体发生癌变时,转铁蛋白受体就会过量表达,以此来转运肿瘤细胞恶性繁殖所需要的过量铁;再如黏蛋白1这种膜结合型糖蛋白,正常情况下,它的作用是保持管道润滑、黏附管腔表面的细菌和病毒等,当机体发生癌变时,这种糖蛋白在细胞膜上的表达量也明显增多。基于抗体识别技术虽具有良好的识别效果,但抗体存在制备步骤繁琐、稳定性差、价格昂贵等特点。因此,发展稳定性好、操作简易、成本低廉、能快速分析糖蛋白的方法对研究相关疾病发生、发展有着重要意义。本论文主要设计制备了两种荧光印迹纳米材料,分别实现了对转铁蛋白和黏蛋白1的特异性识别。具体内容如下:1、以包覆罗丹明B染料的二氧化硅为核,通过溶胶凝胶法在其表面修饰硼酸基团,以辣根过氧化物酶为模板制备了辣根过氧化物酶的印迹聚合物。为了验证方法的普适性,另选转铁蛋白为模板分子,制备了转铁蛋白的印迹聚合物。通过荧光响应考察了印迹材料对模板分子转铁蛋白的识别性能,最后,以转铁蛋白为媒介,实现了细胞膜上转铁蛋白受体的特异性识别与荧光成像分析。该荧光印迹纳米材料具有合成操作简易、识别速度快、稳定性好等优点,更为重要的是,通过对转铁蛋白的特异性识别,实现了对转铁蛋白受体高表达相关循环肿瘤细胞的选择性识别。该工作为科研工作者设计合成人工抗体提供了新思路。2、以上转换纳米材料为荧光团,在其表面修饰硼酸基团。以黏蛋白1为模板,制备了黏蛋白1的印迹聚合物,通过荧光响应考察了印迹材料的识别性能,并将该材料应用于肿瘤细胞膜上黏蛋白1的特异性识别及成像分析研究。最后将其应用于小鼠乳腺癌模型的治疗,通过印迹材料的导向及在980 nm激光照射下的热效应实现对恶性增殖的肿瘤细胞的抑制及杀伤以达治疗肿瘤的目的。实验过程及结果显示,以上转换为荧光团的印迹聚合物具有背景干扰小、操作简便、能够实现肿瘤的识别与治疗一体化等优点。
[Abstract]:Glycoprotein is an important biological macromolecule in organism. It is the product of glycosylation of protein and plays an important role in organism. Studies have shown that the level of glycoprotein expression in organisms is related to many diseases: such as transferrin receptor, a membrane-binding glycoprotein, which binds to transferrin and transports iron into cells to supply normal cell life. When an organism becomes cancerous, the transferrin receptor is overexpressed to transport the excess iron needed for the malignant reproduction of the tumor cell, such as mucin 1, a membrane-binding glycoprotein, normally. Its function is to keep the pipeline lubricated, to adhere to the surface of the tube, bacteria and viruses, when the body carcinogenesis, the glycoprotein expression on the cell membrane is also significantly increased. Although the technology of antibody recognition has good recognition effect, the antibody has the characteristics of tedious preparation, poor stability and high price. Therefore, the method of rapid analysis of glycoprotein is of great significance in the study of the occurrence and development of related diseases. In this paper, two kinds of fluorescent imprinted nanomaterials were designed and prepared, and the specific recognition of transferrin and mucin 1 was realized respectively. The specific contents are as follows: 1. The imprinted polymer of horseradish peroxidase was prepared by sol-gel method with borate group modified on the surface of silica coated with Rhodamine B dye and horseradish peroxidase as template. In order to verify the universality of the method, the imprinted polymer of transferrin was prepared by using transferrin as template molecule. The recognition performance of template molecule transferrin by imprinted material was investigated by fluorescence response. Finally, the specific recognition and fluorescence imaging analysis of transferrin receptor on cell membrane were realized by using transferrin as the medium. The fluorescent imprinted nanomaterials have the advantages of simple synthesis, fast recognition, good stability, and, more importantly, the specific recognition of transferrin. Selective recognition of circulating tumor cells associated with high expression of transferrin receptor was achieved. This work provides a new way for researchers to design and synthesize artificial antibodies. The above conversion nanomaterials are fluorescent groups and modified borate groups on their surfaces. The imprinted polymer of mucin 1 was prepared by using mucin 1 as template. The recognition performance of the imprinted material was investigated by fluorescence response. The material was applied to the specific recognition and imaging analysis of mucin 1 on tumor cell membrane. Finally, it was applied to the treatment of mouse breast cancer model. The tumor cells were inhibited and killed by the guidance of imprinting material and the thermal effect of 980nm laser irradiation. The experimental process and results show that the imprinted polymer transformed into fluorescent group has the advantages of low background interference, simple operation, and the integration of tumor recognition and treatment.
【学位授予单位】：山东师范大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TB383.1;O657.3

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