苯硼酸功能化纳米鱼骨芯片实现肿瘤外泌体高特异性分离分析OA

Early and accurate diagnosis of pancreatic cancer represents a pivotal breakthrough for im-proving patient survival.Although exosome-based liquid biopsy has demonstrated immense promise and attracted widespread attention,existing methods for isolating pancreatic cancer exosomes from blood still face two major bottlenecks:low reaction efficiency within laminar-flow microchips and the lack of tumor-specific exosomal protein markers,which severely compromise both separation specific-ity and efficiency.To tackle these limitations,this study developed a phenylboronic acid-functional-ized three-dimensional nano-herringbone microfluidic chip.By exploiting the covalent binding be-tween phenylboronic acid and the aberrantly overexpressed glycosylated cis-diol structures on tumor exosome surfaces,the chip achieves broad-spectrum,high-affinity recognition of tumor-derived exo-somes.Simultaneously,the periodically ordered nano-herringbone array induces microvortices that disrupt the laminar diffusion boundary layer,substantially enhancing the collision and binding effi-ciency between exosomes and the chip interface.The results demonstrated that the chip achieved a capture efficiency of approximately 85％for exosomes derived from three pancreatic cancer cell lines,PANC-1,SW1990,and PACA-2.In stark contrast,exosomes from normal pancreatic hTERT-HPNE cells exhibited minimal binding,and tumor exosomes subjected to PNGase F-mediated N-gly-can cleavage lost nearly all ability to be captured,definitively confirming that the chip's specific cap-ture is dependent on glycan recognition.Building upon this capability,the chip further integrates in situ enzymatic fluorescence signal amplification,successfully enabling simultaneous molecular profil-ing of six pancreatic cancer-associated membrane protein markers prostate-specific membrane antigen(PSMA),carcinoembryonic antigen(CEA),mucin 1(MUC-1),nucleolin(NCL),protein tyro-sine kinase 7(PTK7),and epithelial cell adhesion molecule(EpCAM)within a single sample.All six markers were significantly overexpressed in tumor-derived exosomes,and distinct cell lines dis-played characteristic expression signatures,demonstrating the chip's capacity to distinguish tumor-derived from normal exosomes and to identify pancreatic cancer molecular subtypes.By integrating glycan-based chemical recognition,efficient fluidic mass transport,and multiplexed molecular pro-filing into a unified platform,this study provides a novel,antibody-independent separation,highly specific,and multi-dimensional information-enriched technology for early pancreatic cancer liquid biopsy,exhibiting significant application prospects in the clinical accurate diagnosis and molecular subtyping of pancreatic cancer.