基于亲水聚合物修饰的新型高比表面积硅基整体柱的制备及在糖蛋白分离富集中的应用

Silica monolithic columns with throughpores and mesopores (bimodal pores) represent attractive alternatives in separation field. However, the monolithic structure and its preparation method have not been improved in recent ten years. In this project, a new synthesis method for bimodal pores silica monolith was put forward. The template induced synthesis method was used to direct bimodal pores simultaneously instead of the post base-treating method as usual. Block polymer was chosen to use as a double-function template to form hierarchically porous silica monolith with both macropores and mesopores, which is a simplification in monolith preparation. The resulted porous material is characterized with a much higher surface area than usual commercial HPLC silica particle, proper mesopore volume and narrow mesopore size distribution, which are admirable in separation. The anti-shrinkage properties make this type of monolith can be made of a variety size of monolithic column. A two step synthesis method using mesoporous membrane deposition on the anti-shrinkage silica column bed was also put forward. The glycoprotein interaction with the stationary phase were studied. By using the monolithic silica columns with high specific surface area and large capacity characteristics, an Mphenylboronic acid functionalized gold nanoparticles, Hydrophilic poly(AAPBA-co-MBA) modified silica monolith and polydopamine boronate silica monolith were prepared. Boronate functional group was covalently immobilized on the monolith via the "thiol-ene" click reaction. The method is novel and reliable, which has a great potential for preparation of different kinds of boronate monoliths. In addition, the monolithic column was further applied to the separation of proteins and glycopeptides, respectively. Ultra-fast and highly efficient separation and enrichment of protein is achieved with minimized sample loss. The successful applications suggested the potential of the new monoliths in glycoprotein analysis.

提出硅源前驱液中添加致双孔模板剂和抗收缩剂的简单方法，以一步法制备高比表面积的适合色谱分离的双孔硅胶整体柱，同时还提出骨架沉积介孔膜法来制备双孔硅胶整体柱的二步合成法。所得柱床的高比表面积、大孔容和窄孔径分布的特性可提供更高的柱效，更凸显整体柱的优势。再利用新型整体柱床的优异特性，根据糖蛋白的分离要求，对柱床进行表面修饰研究，制备出一系列分离新型材料，包括：硼酸纳米金修饰、硼酸基团亲水聚合物修饰、聚多巴胺苯硼酸修饰的硅胶整体柱。引入金纳米粒子，突出其高比表面积、更多活性位点；利用亲水聚合物修饰，凸显其生物相容性和高选择性。拟从亲水功能基团直接热引发自由基共聚，到利用巯基-烯点击反应引入功能基团的两种方法修饰，凸显其反应条件温和、可控；此外还探索更简单有效的新型聚多巴胺苯硼酸整体柱的制备方法，为复杂生物样品中糖蛋白的分离分析提供新的简单高效方法。

硅胶整体材料是一类具有双连续结构和双孔分布的新型材料，并在分离领域展现出良好的性能，提出在硅源前驱液中添加致双孔模板剂和抗收缩剂的简单方法，以一步法制备高比表面积的适合色谱分离的双孔硅胶整体柱，所得柱床的高比表面积、大孔容和窄孔径分布的特性可提供更高的柱效，更凸显整体柱的优势。再利用新型整体柱床的优异特性，根据糖蛋白的分离要求，对柱床进行表面修饰研究，制备出一系列分离新型材料。我们制备了亚氨基二乙酸（IDA）硅基毛细管捕集柱，将亚氨基二乙酸基团接枝到柱体上制备SiO2-IDA毛细管捕集柱。我们发展了一种基于捕集柱的新型柱上富集糖肽的新方法，可作为基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）或纳米LC-MS / MS分析之前的样品预处理步骤，嫁接了两性离子IDA基团后，捕集柱具有出色的亲水性，并且通过亲水作用液相色谱（HILIC）显示出了优异的糖肽柱上富集性能。实验表明此柱上富集方法具有低检出限和高选择性。