结构可控聚电解质刷对蛋白异构体选择性液液凝胶化研究

DelPhi electrostatic modeling will be utilized to visualize the small variance of “charge patches” on protein isoforms. Based on DelPhi modeling, highly charged spherical polyelectrolyte brushes (SPBs) with controllable structures will be designed to integrate electrostatic and hydrophobic interactions in order to mimic the specific interactions of protein epitopes. Multiscale studies on protein isoform selectivity will be performed based on interactions dominated by electrostatics and hydrophobic interaction as a complementarity. Further correlation between protein selectivity and selective phase separation by coacervation will be investigated. This project plans to enhance the binding affinity (Kb) by using highly charged nanoparticles. Meanwhile, through modulation with hydrophobic structures, we hope to amplify the small binding affinity difference among protein isoforms in order to realize highly selective protein adsorption followed by selective protein coacervation with protein activity fully retained. This project challenges the traditional assumption that protein separation based on non-specific electrostatics lacks high selectivity in contrast to affinity interactions and will help to comprehensively understand the electrostatic interactions between SPBs and proteins which significantly contribute to the society of large scale separation of protein isoform by using non-chromatographic method with super high selectivity.

本项目创新性地利用电势模拟软件DelPhi研究高相似蛋白异构体微小的“电荷补丁”分布差异，指导设计具有结构可控高电荷密度的聚电解质刷纳米材料，通过优化聚电解质刷与蛋白质异构体的静电疏水协同作用来模拟亲和色谱中蛋白抗原决定基(epitope)的特异性作用力，从多尺度讨论静电作用主导，短程疏水作用为补充的结构对蛋白异构体选择性的影响，进一步研究选择性吸附与选择性相分离的关联机制。本项目拟通过制备高电荷密度纳米材料以增大亲合力(Kb)，同时辅以疏水改性结构来放大聚电解质刷对蛋白质异构体吸附强度的微小差异，从而在保持蛋白质的活性基础上实现异构体的选择性吸附及大尺度液液凝胶相分离。本项目的开展突破传统思维即非特异性静电作用力缺乏有效的高选择性，建立更加完善的聚电解质刷与蛋白质之间的静电作用理论体系，在利用非色谱法实现超高选择性大尺度分离蛋白质异构体领域有着重要的理论与工程实践意义。

利用电势模拟软件DelPhi研究高相似蛋白异构体微小的“电荷补丁”分布差异，制备了一系列具有尺寸、亲疏水性、形貌及电荷可控高电荷密度的聚电解质刷纳米材料，通过优化聚电解质刷与蛋白质异构体的静电疏水协同作用来模拟亲和色谱中蛋白抗原决定基(epitope)的特异性作用力，从围观和宏观尺度讨论静电作用主导，短程疏水作用为补充的结构对相似蛋白质选择性的影响，通过热力学参数的测定，研究选择性吸附与选择性相分离的关联机制。本项目通过制备高电荷密度纳米材料以增大亲合力(Kb)，同时辅以疏水改性结构来放大聚电解质刷对蛋白质异构体吸附强度的微小差异，从而在保持蛋白质的活性基础上实现异构体的选择性吸附。本项目的开展突破传统思维即非特异性静电作用力缺乏有效的高选择性，建立更加完善的聚电解质刷与蛋白质之间的静作用理论体系，在利用非色谱法实现超高选择性大尺度分离蛋白质异构体领域有着重要的理论与工程实践意义。项目进行过程中总共发表标注基金的SCI论文19篇，授权专利2篇。