【作者】 陆书来；

【导师】 成国祥；

【作者基本信息】 天津大学 ， 材料学， 2003， 博士

【摘要】 分子印迹技术是从仿生角度，采用人工方法制备对模板分子具有专一性结合作用的聚合物的技术。分子印迹聚合物在分离提纯、免疫分析、酶模拟以及生物模拟传感器等许多方面显示出广泛的应用前景。本文对分子印迹的基本原理、分子印迹聚合物微球的制备方法和应用、蛋白质分子印迹的特点和方法，以及磁性复合微球的制备方法和应用进行了较为详细的总结和评述。分别采用悬浮聚合法和反相乳液—悬浮聚合法制备了氨基酸分子印迹聚合物磁性复合微球（MS-SMIPs）；分别采用反相悬浮聚合法和种子反相悬浮聚合法制备了蛋白质分子印迹磁性软湿凝胶复合微球（MS-PIGMs）；用扫描电子显微镜、热重分析仪、振动样品磁强计、红外光谱、高效液相色谱等对所制备的MS-SMIPs和MS-PIGMs进行了表征。研究表明，悬浮聚合法MS-SMIPs制备工艺简单，但不适合高粘度单体；反相乳液—悬浮聚合法通过反相乳液对Fe3O4粒子的预包埋，解决了Fe3O4粒子与油溶性单体和聚合物的相容性问题，并解决了高粘度单体在普通悬浮聚合中难以分散成球的问题，制得的MS-SMIPs内部有大量空腔，因而降低了自身密度，在水相识别体系中不易沉淀；反相悬浮聚合法所制备的MS-PIGMs，因其交联网络和凝胶孔穴可根据环境的变化而改变，对模板蛋白质的洗脱和吸附十分有利；种子反相悬浮聚合法制备的MS-PIGMs，实现了蛋白质的微球表面印迹，解决了蛋白质包埋法中因模板无法洗脱干净而造成的模板蛋白质利用率较低的问题，同时提高了蛋白质在凝胶微球中的传质和扩散速率。研究结果表明，所制备的MS-SMIPs形态均为较好的球形，并均具有一定的磁响应性；搅拌速度、分散剂用量、Fe3O4加入量等对MS-SMIPs的粒径和形态具有明显的影响；酪氨酸和色氨酸印迹的MS-SMIPs对模板分子均具有较高的分子识别性能，而苯丙氨酸印迹的MS-SMIPs对模板分子无明显的分子识别性能。所制备的MS-PIGMs形态也为较好的球形，湿态时粒子表面具有大量分布较为均匀的孔穴，干态时因粒子收缩表面孔穴闭合；分散剂、引发剂、Fe3O4用量等均对MS-PIGMs形态有明显的影响；所制备的MS-PIGMs具有一定的磁响应性，并具有较好的分子识别性能，其对模板的识别主要靠印迹孔穴与模板蛋白质分子因非常近的表面接触而产生的众多氢键的协同作用。更多还原

【Abstract】 Molecular Imprinting Technique (MIT) is a technique originated from bionic to prepare polymers with recognition specificity to template molecules by manual method. Molecular Imprinting Polymers (MIPs) have exhibited extensive application prospect in separation, purification, immunoassay, enzyme mimic, biomimic sensor, and other fields.In this thesis, the principle of MIT, the preparation methods of molecularly imprinted polymeric microspheres (MIPMs, SMIPs), and the application of MIPs, as well as the specialty and methods of protein template imprinting were expatiated. The preparation methods and the applications of magnetic composite microspheres (MCMs) were reviewed in detail. Amino acid molecules imprinted polymeric composite microspheres with magnetic susceptibility (MS-SMIPs) were prepared by suspension polymerization (SP) and inverse emulsion-suspension polymerization (IESP), respectively. Protein molecules imprinted soft-wet gel composite microspheres with magnetic susceptibility (MS-PIGMs) were prepared by inverse suspension polymerization (ISP) and seeded inverse suspension polymerization (SISP), respectively. The resulting MS-SMIPs and MS-PIGMs were characterized by scanning electron microscope (SEM), thermogravimetric analyzer (TG), vibrating sample magnetometer (VSM), infrared spectrum (FTIR), high performance liquid chromatography (HPLC), and so on.It was showed that SP method was easier than IESP method being used to prepare MS-SMIPs, but SP method didn’t suitable for high viscosity monomers to be used to prepare MS-SMIPs. The compatibility of Fe3O4 particles with monomers and polymers was solved through pre-encapsulation of Fe3O4 particles by inverse emulsion in IESP method, the difficulty for SP method to disperse high viscosity monomers to turn into droplets was overcome as well, and precipitation didn’t occur easily in aqueous recognition system, for the resulting MS-SMIPs had a large quantity of air cores inside, hence had low density themselves. The<WP=6>resulting MS-PIGMs by ISP method could adjust their crosslinked network and gel pore size themselves, being helpful to elution and adsorption of proteins. MS-PIGMs prepared by SISP method realized microbead surface imprinting of protein molecules, so the problem of low utilization ratio of template proteins caused by residual of templates in "protein entrapment imprinting" method was solved, while the rate of mass transfer and diffusion of proteins inside MS-PIGMs were improved.The results showed that the resulting MS-SMIPs were all good spheroid form, and had certain magnetic responsibility. Stirring rate, amount of dispersant and Fe3O4, and other factors all could affect the size and morphology of the resulting MS-SMIPs obviously. Tyrosine imprinted MS-SMIPs and tryptophan imprinted MS-SMIPs had higher molecular recognition specificity to their templates, respectively. Phenylalanine imprinted MS-SMIPs had no obviously molecular recognition specificity in contrast. The resulting MS-PIGMs were all spheroid form, and had large quantity of regularly distributed pores, which close in dry condition. Dispersant, initiator, Fe3O4, and other factors could all affect the morphology of the resulting MS-PIGMs. MS-PIGMs all had certain magnetic responsibility, and had higher molecular recognition specificity to their templates. The recognition of MS-PIGMs to their template proteins was due to the synergic action of a large number of hydrogen bonds caused by a very close surface contact between a template protein and the imprinted cavity.更多还原