几种新型表面分子印迹材料对糖蛋白的分离与检测

发布时间：2018-06-19 13:06

本文选题：糖蛋白 + 表面分子印迹聚合物　；参考：《湘潭大学》2017年硕士论文

【摘要】：糖蛋白作为多种疾病的生物标志物,对临床诊断具有巨大意义。然而,在没有任何预处理的复杂生物样品中测定低丰度糖蛋白仍然是一个问题。本文提出了基于表面分子印迹和智能材料的方法用于分离和检测糖蛋白。主要研究内容如下:1、提出了一种基于阳离子p H敏感水凝胶和硼酸材料的磁性纳米微粒的新型分子印迹技术用于分离和富集糖蛋白。选用4-乙烯基苯硼酸(VPBA),甲基丙烯酸二甲氨基乙酯(DMA)和N-N-亚甲基双丙烯酰胺(MBA)合成印迹层。通过测试不同p H值下的溶胀比表征了印迹材料受p H调控的溶胀/收缩性能,同时也对该材料对目标糖蛋白的吸附性能进行了研究。结果表明,模板分子的洗脱和吸附能受p H调控,而且能在10min达到81.2mg/g的吸附量。2、用修饰苯硼酸的磁性Fe_3O_4纳米微粒为载体,首先利用硼酸亲和键与糖基的共价键结合力使得目标糖蛋白OVA定轨在载体的表面。多巴胺为单体,自聚合反应形成包覆在载体表面的印迹层,最后去除模板,得到对OVA有特异性吸附的磁响应分子印迹纳米颗粒。同以上体系,做了一系列表征和条件优化。结果表明,制备的印迹粒子对OVA有较好的吸附能力和选择性能。3、制备了一种新型共振光传感器检测糖蛋白,具体合成过程与体系一类似。在分子印迹过程中,印迹层中形成了对目标糖蛋白有特异性识别的空腔。对目标分子地选择性捕获对应着传感器RLS强度的变化。该传感器具有效率高(15min)和检出限低(0.13 n M)的优点。
[Abstract]:As a biomarker of many diseases, glycoprotein has great significance in clinical diagnosis. However, the determination of low-abundance glycoproteins in complex biological samples without any pretreatment remains a problem. A method based on surface molecular imprinting and smart materials is proposed for the separation and detection of glycoproteins. A novel molecular imprinting technique based on cationic pH sensitive hydrogels and boric acid materials was proposed for the separation and enrichment of glycoproteins. The imprinted layer was synthesized by using 4-vinylphenylborate (VPBA), dimethylaminoethyl methacrylate (DMA) and N-N- methylene bisacrylamide (N-N- methylene bisacrylamide). The swelling / shrinkage properties of the imprinted materials regulated by pH were characterized by measuring the swelling ratios at different pH values. The adsorption properties of the imprinted materials to the target glycoproteins were also studied. The results show that the elution and adsorption of the template molecule can be regulated by pH, and the adsorption capacity of 81.2mg/g can be reached by 10min. The template molecule can be supported by magnetic Fe _ 3O _ 4 nanoparticles modified with phenylboric acid. Firstly, the covalent binding force of boric acid affinity bond with glycosyl group makes the target glycoprotein OVA orbit on the surface of the carrier. Dopamine was used as monomer and self-polymerized to form imprinted layer coated on the surface of carrier. Finally, the template was removed to obtain magnetically responsive molecularly imprinted nanoparticles which were specifically adsorbed to OVA. In the same system, a series of characterization and conditions were optimized. The results showed that the imprinted particles had good adsorption ability and selectivity to OVA. A new resonance light sensor was prepared for the detection of glycoproteins. The synthesis process was similar to that of the system. In the process of molecular imprinting, a cavity with specific recognition of target glycoprotein was formed in the imprinting layer. The selective capture of the target molecule corresponds to the change of the RLS intensity of the sensor. The sensor has the advantages of high efficiency (15 min) and low detection limit (0.13 nm).
【学位授予单位】：湘潭大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O631.3;TQ937

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