核苷酸稀土配位聚合物的设计合成及凝血酶识别传感研究

In this project, nucleotides and lanthanide ions are screened and designed as components to form amorphous nucleotides-based lanthanide coordination polymer nanoparticles. Based on the study of self-assembly mechenism between biomolecular ligands and lanthanide ions as well as adaptive property of the formed coordination polymer, multifunctional lanthanide coordination polymer hybrid nanoparticles with unique features, such as high fluorescence quantum yield, large Stokes shift, and good biocompatibility can be constructed by adaptive inclusion of guest materials such as dyes, metal nanoparticles, and fluorescent nanomaterials in the course of self-assembly of nucleotides and lanthanide ions. Based on the special interaction between thrombin and lanthanide ions, the effect of thrombin on mophology, structure, and fluorescence behavior of lanthanide coordination polymer nanoparticles, as well as the binding site, coordination number, and mechanism between thrombin and lanthanide ions will be studied in detail. The thrombin-special recognization process can be effectively converted into spectral signals of lanthanide coordination polymer nanoparticles. Based on this, sensing strategies for highly sensitive and selective assay of thrombin activity can be developed by taking advantages of the fluorescence, multi-mode sensing, and visible features of the multifunctional lanthanide coordination polymer nanoparticles. Furthernore, the sensing platform will be used to detect thrombin activity in biological samples. The further investigation on the interaction between inhibitors and thrombin provides new ideas for targeted drug screening.

本项目拟以核苷酸及其衍生物等天然生物分子和稀土离子为组装基元制备稀土配位聚合物，在研究生物分子配体与稀土离子的自组装机理以及自适应特性的基础上，实现多种类客体材料在聚合物中的有效包络，制备荧光量子产率高、Stokes位移大、生物相容性好的多功能稀土杂化纳米材料。基于凝血酶与稀土离子之间的特殊相互作用，研究凝血酶对稀土纳米材料的形貌、结构以及光谱行为的影响，将凝血酶识别过程有效转换为稀土纳米材料的光谱信号变化，研究稀土配合物与凝血酶之间的反应动力学过程，揭示凝血酶与稀土离子的结合位点、作用方式及机理，构建基于多功能稀土杂化纳米材料多信号输出策略的凝血酶活性检测新方法，并应用于实际样品中凝血酶活性的检测。进一步研究凝血酶活性与其抑制剂分子结构之间的内在关系，为凝血酶靶向药物的设计筛选提供新思路。

稀土配位聚合物具有荧光量子产率高、Stokes位移大、生物相容性好等特性，其发光性能研究受到了极大关注。本项目制备了一系列新型稀土配位聚合物以及多功能稀土杂化纳米材料，构建了系列基于稀土配合物多信号输出策略的凝血酶检测新方法，并探索在酶及金属离子分析中的应用。（1）以稀土Tb3+为中心离子及鲁米诺为配体分子合成稀土配合物荧光探针，基于凝血酶对荧光探针的聚集诱导发光效应，构建了荧光及可视化多信号输出策略检测凝血酶。（2）稀土离子Er3+与荧光石墨烯量子点配位可使量子点聚集而荧光减弱，构建了基于Er3+介导石墨烯量子点荧光开关的凝血酶传感方法。（3）以稀土Tb3+为发光中心离子与富G碱基的凝血酶适配体配位合成稀土配合物荧光探针，基于凝血酶对探针荧光的猝灭作用建立了凝血酶分析方法并探索识别机理。（4）以凝血酶适配体为模板制备兼具适配体结构功能以及荧光特性的荧光量子点，在凝血酶作用下量子点/AuNPs聚集，构建了比色、荧光、光散射等多信号输出的凝血酶分析方法。（5）以Tb3+为中心离子、GMP为配体、量子点为客体材料合成稀土荧光纳米材料，构建比率荧光和可视化多信号输出方法检测As(V)。（6）以Tb3+为中心离子、GMP为主配体及鲁米诺为辅助配体合成双配体稀土配合物比率荧光探针，构建了比率荧光法及可视化法检测ALP和As(V)。（7）以Eu3+为中心离子、鲁米诺和GMP为双配体分子、硫黄素T为客体分子，制备同时发射鲁米诺和硫黄素T荧光的稀土荧光探针，构建比率荧光法检测ALP和As(V)。（8）合成了同时发射Eu3+和鲁米诺荧光的稀土荧光探针，基于Hg2+对鲁米诺荧光的抑制和对Eu3+荧光的增强作用，构建了比率荧光法和比色法检测Hg2+。（9）通过Ce3+与2,5-二甲巯基-对苯二甲酸配位合成稀土铈配合物荧光探针，构建了基于聚集诱导荧光猝灭效应的As(III)分析方法。（10）合成了铈铽双稀土配合物荧光纳米材料，构建了基于ALP催化P2O74-水解的ALP检测及其抑制剂筛选方法。本项目已按计划完成，发表SCI论文49篇，授权发明专利13项，申请发明专利3项。