蛋白印迹响应性高分子磁性复合微球多层次结构调控与性能

For the problems of the difficulty in the protein imprinting, we carry out the theoretical research on the multi-level structure, regulation and application of environment-responsive polymer magnetic microspheres for protein imprinting recognition. The specific interaction of template proteins and environment-responsive polymers is investigated, and the relationship between polymer chain structures, cross-linked structures, electricity charge, the site and number of functional group and the selective recognition property for template proteins is established; the ways of utilizing PMPC to enhance the selective property of imprinted microspheres to template proteins are explored; the methods for regulating the surface morphologies and large pore structure of imprinted microsphere to improve the specific surface area of imprinted microsphere is studied, and the relationship between surface morphologies, structure of pores and the efficiency of imprinting is revealed. A series of protein imprinted composite microspheres with different interface characteristics are built and the influence of interface structures such as rattle-type to protein separation was explored. The adsorption and desorption mechanisms and dynamic behavior in the imprinting layer of template protein are all revealed, and the corresponding theoretical models are built. Finally the controllable preparation of the space of imprinted cavity and the point arrangement can be realized through the levels of polymer structure, cross-linked structure and pore structure. By solving such key problems as the limition in the selective recongnition of protein imprinting materials and the capture and release of target molecules, we lay a new way of thinking for the researches of extraction, enrichment, immunoassay and the drug control release of biomacromolecules.

针对蛋白等生物大分子难于印迹的问题开展系统的蛋白印迹环境响应性高分子磁性微球多层次结构构筑、调控与应用的理论研究。探索模板蛋白和环境响应性高分子可能的特异性相互作用，建立聚合物链结构、交联结构、电荷和官能团位点与数量等与模板蛋白分子选择性识别性质之间的关系；探索出利用PMPC等增强印迹微球对模板蛋白选择性的途径；研究印迹微球表面形貌及大孔道调控方法，提高印迹微球的比表面积，揭示表面形貌、孔结构和印迹效率的关系；构筑具有不同界面特征的系列蛋白印迹复合微球，探索摇铃型印迹微球等界面结构对蛋白分离效果的影响规律；揭示模板蛋白吸附和解吸过程机制及其在印迹层中的动力学行为，建立对应的理论模型；最终从高分子分子结构、交联结构、孔道结构等多层面实现对印迹腔、位点排布的可控制备；解决制约蛋白印迹材料的选择性识别和目标分子捕获释放等关键问题，为生物大分子的分离、富集、免疫检测、药物控制释放等研究提供新思路。

由于蛋白等生物大分子自身结构特点，其分离纯化是最具挑战性的难题之一。本项目围绕模板蛋白和环境响应性高分子之间的相互作用、聚合物多级结构对蛋白选择性识别等科学问题，开展了系统研究，相关成果在ACS Applied Materials & Interfaces，Macromolecules等期刊上发表49篇论文，他引645次，授权专利11项，获2017年国家科技进步奖二等奖1项。主要成果有：.（1）制备了系列磁性温敏印迹微球，揭示了印迹层厚度、界面羧基密度等对BSA识别效果的影响规律，以及温敏性能对蛋白吸附与洗脱的作用，探索出交联度、温敏性和识别能力间的平衡关系；.（2）将系列离子液体以及离子液体功能化的线性聚合大分子链作为功能单体，利用分子模拟揭示其与蛋白间的相互作用，利用可控聚合制备出系列印迹微球，有效解决了印迹过程中模板蛋白易失活变性的问题；.（3）构筑了多种新型蛋白印迹聚合物体系：集成Ni2+-BSA定向配位作用和分子间氢键等作用，制备的多层次结构磁性蛋白印迹微球实现了吸附量和选择性双提升（Q=266.99 mg/g，IF=5.45）；将硫代内酯点击开环聚合多步反应策略引入磁性蛋白印迹体系，实现了印迹高分子结构的灵活设计，获得了兼具高效吸附和识别性能的印迹材料；.（4）利用共聚合和AGET-ATRP法将具有抗蛋白吸附性质的甲基丙烯酰乙基磺基甜菜碱、2-甲基丙烯酰氧乙基磷酰胆碱等可控引入印迹体系，通过精确控制其链段长度、排列方式等，实现了印迹层表面抗蛋白吸附能力的有效调控，显著提高了特异性识别能力，印迹因子（IF）增至8.32；.（5）基于受限自缩聚反应制备出具有自驱动性能的表面印迹管状碳纳米纤维，其兼具高吸附量（541.99 m/g）与快吸附的特性；发现了非交联PGMA种子粒子具有单洞特性，构筑了多种树莓结构微球，提高了印迹表面积。基于这些微球制得的高比表面印迹材料具有优良的吸附性能（IF>4.5）。. 本项目从高分子分子结构、交联结构、孔道结构等多层面实现了对印迹腔、位点排布的可控制备，为生物大分子的分离富集、免疫检测、药物控释等研究提供新思路。