外泌体是近年来备受关注的液体活检标志物。外泌体与其他细胞外囊泡因性质相似而难以精准分离，且外泌体运载的某些疾病标志物丰度极低而难以检测，严重限制了其在液体活检中的应用。本项目计划开发多孔纳米鱼骨芯片，对外泌体实现高精准分离和高灵敏检测。多孔纳米鱼骨芯片通过碳纳米管和聚苯乙烯球的介导自组装再洗去致孔剂制备，具有孔径均一（150 nm）的多孔结构和外泌体捕获抗体，实现体积排阻+免疫亲和的双参数正交捕获，显著提高外泌体分离的纯度；三维多孔纳米鱼骨结构增强流体的混匀效果，同时提供大的表面积，提高芯片界面反应效率，实现外泌体标志物的高灵敏定量分析。基于多孔纳米鱼骨芯片，本项目对颅脑损伤的外泌体标志物SYP、C5b-9、CR1和CD59进行高灵敏、多通道的分析，并开发基于脑脊液外泌体的颅脑损伤快速诊断新方法。该项目的实施将不仅推动外泌体分离分析技术的进步，而且为颅脑损伤的快速诊断提供液体活检新策略。

Exosomes are thought to be ideal markers for liquid biopsy. However, it’s still technical challenging to isolate exosomes from extracellular vesicles selectively and quantify low abundance biomarkers of exosomes sensitively, which limits the application of exosomes in liquid biopsy severely. Herein, we propose to develop a porous nano-herringbone chip aiming to isolate exosomes precisely and detect exosomal biomarkers sensitively. The porous nano-herringbone chip is prepared by self-assembly of carbon nanotubes and polystyrene beads, and affords uniform porous structures (150 nm) and exosome capture antibodies, thus adopts the double-parameter orthogonal capture principle of size-exclusion and immunoaffinity to achieve highly purified exosomes. Moreover, 3D porous nano-herringbone structures enhance the fluidic mixing effect significantly and provide huge surface area, enabling highly sensitive analysis of exosomal markers. Using this porous nano-herringbone chip, we detect several exosomal markers related to traumatic brain injury (SYP, C5b-9, CR1 and CD59) in a sensitive and multiplexed way, and further apply to early diagnosis of traumatic brain injury. The proposed project will not only promote the development of exosome isolation and detection technologies greatly, but also develop a new liquid biopsy strategy for early diagnosis of traumatic brain injury.