光/磁纳米粒子复合组装体的可控构筑及其对外泌体的高效特异性分离

In order to highly identify and bio-separate the exosomes specifically, the project is aimed to design and construct a kind of self-assemblied magnetic composite with selectively efficient recognition, fluorescent real-time monitoring and external stimulus response properties. Taking use of self- assembly technology, through the host-guest interaction and the hydrophobic interaction between the amphiphilic superparamagnetic magnetic nanoparticles and quantum dots, to cooperative prepare the supramolecular hybrids, and to realize the controllable preparation of self-assembling behavior and fine micro structure at the base of multilevel and multifunction. The fluorescently labeled different antibody is chemically bonded to the hybrids by a photocleavable linker respectively to realize specific recognition and efficient bio-separation for exosomes from different cell sources. The controllable pro-preparation techniques and methods for the two kinds of inorganic nanoparticles will be established. The hierarchical self-assembly mechanism and rules to form the controllable structure and functions will be elucidated, through host guest interaction and hydrophobic interaction. Furthermore, the project will reveal the relationship between the antibody coupling, capturing and separation efficiency and the external stimuli etc., which will further develop the applications of magnetic nano-biomaterials in biomedicine.

本项目针对外泌体的高特异性识别和分离，拟设计构建一种集选择性高效识别、荧光实时监测、外界刺激响应的磁性复合组装体。利用分子自组装技术，通过两亲性超顺磁性纳米粒和两亲性量子点间的主客体作用及亲疏水作用进行二元协同自组装，制备出多层次、多功能的超分子杂化组装体，实现自组装行为和精细微结构的可控制备。通过键合荧光标记的特异性抗体的光致化学键断裂偶联剂进行表面适当功能化，实现对不同细胞来源的外泌体特异性识别和快速分离。建立表面性质可控的两亲性无机纳米粒子组装单元的制备技术和方法；阐明通过主客体作用和亲疏水作用的分子多级自组装，形成结构和功能可控的复合组装体的影响机制和规律；揭示复合组装体的表面抗体偶联等功能化对外界刺激响应的能力、外泌体的捕获及其分离效率等行为的调控和作用规律，进一步开发和拓展磁性纳米生物材料在生物医学中的应用。

外泌体因其特有的结构和所荷载的完整的母细胞来源的特异性蛋白以及遗传性物质而成为疾病早期临床诊断和治疗的有效标志物。本研究通过主客体自组装技术，构建基于二元功能磁性纳米粒子的自组装平台BIS-NPs，实现了在复杂生物样本（完整细胞培养液和体液）中外泌体的高特异性和快速磁分离以及温和条件下的可控释放。与传统的超高速离心法以及商用试剂盒方法相比，该平台可以高效且高纯度地富集外泌体，并保留其良好的结构和功能完整性，从而为下游生物学分析提供便利。