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摘要磁性聚合物微球是一种兼具磁性材料和高分子材料特点的核壳结构磁性微球，由于其比表面积大，表面基团多，生物相容性好，已经被应用在药物输送和液体活检等众多领域，本研究在参考众多文献的基础上，合成了性能优异的聚苯乙烯磁性微球并用来分离提纯 mRNA。本论文的主要研究内容有： 1.利用共沉淀法并选择油酸作为表面修饰剂制备了油溶性的磁性纳米微粒，并利用XRD、TEM和FT-IR等表征手段对该磁性粒子进行表征，产物粒径均一，磁响应性优异，具有超顺磁性。 2.苯乙烯和丙烯酸在制备好的磁性纳米粒子表面上进行共聚，对合成的磁性聚合物微球进行表征，产物表面修饰上羧基，粒径达到微米级，磁场下较快分离，具有超顺磁性。 3.磁性聚合物微球表面偶联 dT，分离纯化 mRNA，利用紫外可见分光光度计进行定量和定性分析，合成微球可用来分离 mRNA。 41663
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毕业论文关键词 化学共沉淀法 聚苯乙烯磁性微球 分散聚合法 mRNA分离
Title Synthesis of magnetic beads and their application for mRNA isolation
Abstract Magnetic polymer microspheres (MPM) are a kind of novel functional materials, which exhibit properties of magnetic materials and polymer materials. MPM has been used in biological system because of their large specific surface area, good biocompatibility and so on. In this paper, magnetic polystyrene microspheres have been synthesized and used for mRNA isolation on the base of others’ research. The main content of this paper is such as following: 1. We prepared magnetic nanoparticles with the chemical co-precipitation method and oleic acid as modified surfactant. The synthesized nanoparticles were characterized by XRD, TEM and FT-IR, which exhibit uniform sizes and good magnetic response. 2. Polystyrene magnetic microspheres are prepared by dispersion polymerization method which used styrene and acrylic acid. The synthesized beads were characterized, which exhibit good magnetic response, carboxylation and micro-size. 3. NH2-labeled oligo-(dT) 20 were immobilized on magnetic microspheres and used for mRNA isolation. We used UV-VIS spectrophotometer to demonstrate the isolation of mRNA.
Keywords chemical co-precipitation method magnetic polystyrene microspheres dispersion polymerization method mRNA isolation
目次
1绪论.1
1.1mRNA分离技术简介1
1.1.1寡聚（dT）-纤文素柱层析.1
1.1.2寡聚（dT）-磁性微球法分离纯化mRNA2
1.2磁性复合微球.3
1.2.1纳米Fe3O4磁性微粒的制备3
1.2.2磁性高分子微球的合成.6
1.3本文的研究思路和研究内容8
2实验部分9
2.1试剂与仪器.9
2.1.1实验试剂9
2.1.2实验仪器9
2.2Fe3O4磁性纳米粒子的制备10
2.3油酸改性的Fe3O4纳米粒子的制备.10
2.4聚苯乙烯包覆磁性粒子的制备11
2.5探针偶联修饰及其提取信使RNA的研究12
2.6测试与表征.12
3结果与讨论14
3.1油酸改性的Fe3O4磁性微粒的表征.14
3.1.1Fe3O4磁性纳米粒子的X射线衍射图.14
3.1.2Fe3O4磁性纳米粒子的红外谱图.15
3.1.3Fe3O4磁性纳米粒子的热重分析图.17
3.1.4Fe3O4磁性纳米粒子的磁滞回线图.17
3.1.5Fe3O4磁性纳米粒子的透射电镜图.18
3.1.6Fe3O4磁性纳米粒子的分散稳定性测试19
3.2聚苯乙烯磁性微球的表征20
3.2.1聚苯乙烯磁性微球的X射线衍射图.20
3.2.2聚苯乙烯磁性微球的红外谱图.20
3.2.3聚苯乙烯磁性微球的磁滞回线图.21