基于磁性金属有机骨架材料的低丰度蛋白/肽段高效选择分离分析新方法研究

Low abundance biomolecules has already taken a key place in clinical disease biomarkers screening, clinical diagnosis and therapy, etc. Therefore, it is necessary to develop new techniques and methodologies for the rapid and effective sample analysis. Metal organic frameworks (MOFs) have become one of the most exciting research topics and attracted considerable interest in recognition, catalysis, separation and adsorption because of its outstanding ultrahigh porosity, enormous internal surface areas and chemical properties. Especially, the precisely chemical modification can endow the specific functions without changing the underlying topology to afford a possibility for the bio-separation and technique. In this proposal，by combining magnetic physical technology, nanotechnology, surface functionalization, and chemical derivatization with traditional bioanalytical methods, a series of newly-designed bioanalytical methods to resolve existing problems in specific enrichment and detection biomolecules has been developed. This proposal made full use of the synergistic effect of magnetic MOFs from the two components of magnetic properties, highly ordered pores and special surface properties, through postsynthetic modification to synthesize several functional magnetic MOFs for different research requirements, and novel technologies have been introduced into low abundance proteome research fields. Furthermore, bio-tagging quantitative technology is combined for study the new application of magnetic MOFs in high selective separation, enrichment and quantitation of low abundance proteins and peptides. Finally, we aim at exploring and finding out widespread methodology in the selective enrichment and quantitation of target biomolecules. The application of MOFs in bio-analysis fields will not only open up a new possibility for the rapid analysis but also extend the application of MOFs mesoporous material.

低丰度生命分子研究已在临床标志物筛选和早期诊断领域发挥重要作用，而发展新技术与样品处理方法对于其快速高灵敏分析至关重要。金属有机骨架材料（MOFs）因其迷人的框架结构、高孔隙率等特性，广泛用于识别、催化、分离吸附等领域，而通过精确地可控设计可以在不改变其拓扑结构的前提下赋予其新的功能，为解决现存的生物分离、分析技术难题提供了新的切入点。本项目拟将磁性物理技术、材料技术、表面功能化及化学衍生与传统分析方法相结合，发展一系列生物样品高效特异性富集分析新方法。充分利用磁性MOFs的磁分离与高比表面积协同作用，通过可控的特定识别基团合成后修饰，制备适用于不同研究目的的新型功能磁性MOFs，为解决低丰度蛋白/肽段的特异性富集提供新颖的研究手段。并结合生物标记定量技术，集中于开发新型功能化磁性MOFs在低丰度目标分子高选择性分离富集及定量分析中的新应用，探索普适性的生命分子选择分离富集定量分析新策略。

疾病相关低丰度生物标志物的筛选已在早期临床诊断领域发挥重要作用，而发展新技术与样品处理方法对于其快速高灵敏分析至关重要。本项目以低丰度蛋白、多肽和核酸等生命分子为研究对象，将材料技术、化学衍生及生物放大等多种手段与传统分析方法相结合，发展了一系列低丰度生物标志物的高灵敏、高选择分离及分析体系，研究工作包括以下几部分：（1）设计合成了几种新型多功能金属有机骨架材料（UiO-66、FeTCPP⊂UiO-66、Pt/UiO-66、Fe3O4@UiO-66、Fe3O4@MIL-100(Fe)、Bubble@ (MIL-100(Fe) 等），通过可控的特定识别基团合成后修饰，制备出了适用于不同研究目的的新型功能MOFs， 探讨了其替代基质功能及用于磷酸化翻译后修饰研究的分析效果及机理探讨。（2）基于靶诱导耦合生物条形码纳米放大技术，DNA酶组装及均相外切酶III辅助目标循环放大等多重技术，探索设计构建了几个低丰度生物标志物的比例放大双信号平台，用于凝血酶和口腔癌DNA等生物标志物传感分析，并进一步拓展构建了新颖的均相比例质谱动态可调酶活性检测平台。（3）基于双酶放大和浮选分离耦合比例放大，分别构建了界面和均相体系的循环肿瘤DNA检测平台。上述研究工作充分利用了现有的分离分析手段，实现了对多种低丰度生物标志物的灵敏、快速检测，取得了具有重要学术意义和应用参考价值的研究成果。发表SCI论文13篇，影响因子大于5.0的7篇。