分析化学2026，Vol.54Issue(3)：339-351,中插1-中插2,15.DOI:10.19756/j.issn.0253-3820.251351
上转换发光纳米探针协同滚环扩增用于胰腺癌外泌体表型分析
Upconversion Luminescent Nanoprobes in Conjunction with Rolling Circle Amplification for Phenotypic Profiling of Exosome from Pancreatic Cancer
摘要
Abstract
Exosomes have emerged as potential biomarkers for non-invasive diagnosis and real-time monitoring of disease due to their ability to carry bio-molecular information from their source cells and stability in various body fluids.However,existing detection methods face challenges of insufficient specificity,limited sensitivity,and difficulty in distinguishing exosomes from different cellular origins.To address this,this study developed a fluorescent biosensing platform based on silica-coated upconversion nanoparticles(UCNPs@SiO2)for characterization of exosomes derived from pancreatic cancer cells that simultaneously overexpress epidermal growth factor receptor(EGFR)and epithelial cell adhesion molecule(EpCAM).The platform employed a cascade signal amplification strategy.Firstly,exosomes were captured using magnetic beads conjugated with anti-CD63 antibodies.Subsequently,specific recognition of EGFR and EpCAM was achieved via DNA aptamers,triggering rolling circle amplification(RCA)to generate long-chain DNA products containing repetitive sequences.Finally,UCNPs-labeled DNA probes complementary to RCA products were added.After incubation and magnetic separation,the fluorescent signal of free UCNPs in the supernatant was detected.Experimental results showed that the supernatant fluorescence intensity decreased with increasing exosome concentration,exhibiting a negative correlation response.Additionally,semi-quantitative naked-eye analysis could be performed by observing the degree of magnetic bead aggregation,with the lowest visually distinguishable concentration for EGFR-and EpCAM-positive exosomes being 3.0×103 particles/mL and 6.0×103 particles/mL,respectively.The analytical method established in this study enabled high-sensitivity,dual-protein phenotyping of exosomes from specific cellular origins,providing a novel methodological support for non-invasive diagnosis and therapeutic monitoring of pancreatic cancer.关键词
外泌体/表型分析/上转换发光纳米材料/滚环扩增/胰腺癌Key words
Exosome/Profiling/Upconversion nanoparticles/Rolling circle amplification/Pancreatic cancer引用本文复制引用
董青,孙旭,李丹,汪尔康,汪劲..上转换发光纳米探针协同滚环扩增用于胰腺癌外泌体表型分析[J].分析化学,2026,54(3):339-351,中插1-中插2,15.基金项目
吉林省中青年科技创新创业卓越人才(团队)项目(No.20240601015RC)和山东省自然科学基金项目(No.ZR2023QB291)资助. Supported by the Scientific and Technological Development Project of Jilin Province(No.20240601015RC)and the Natural Science Foundation of Shandong Province Youth Project(No.ZR2023QB291). （团队）
