Development of plasmon-enhanced biosensing for multiplexed profiling of extracellular vesicles

细胞外囊泡（EV）为非侵入性液体分子诊断提供了新的机会 活组织检查这些细胞衍生的膜结合囊泡大量存在于生物流体中。电动汽车携带 细胞特异性货物（例如，脂质，蛋白质和遗传物质），可以利用它们来探测 它们细胞起源的分子状态。然而，电动汽车分析由于电动汽车的 纳米尺寸和存在于广阔的生物背景中。然而，EV分析提出了独特的技术 由于电动汽车的纳米尺寸和存在于广阔的生物背景中，这是一个挑战。而新 虽然已经开发了EV分析技术，但基本的限制仍然存在，包括i）低 灵敏度限于批量分析; ii）需要EV裂解以检测EV内的标志物; iii）缺乏 对蛋白质和RNA标记物的多重分析;以及iv）在分离前需要单独的EV分离过程。 比色法本申请的总体目标是克服这些技术挑战并开发新的 该平台能够对单个EV中的EV蛋白和RNA标记物进行多重分析。我们之前 开发了基于透射表面等离子体共振的纳米等离子体EV传感平台， 周期性纳米孔光栅我们发现纳米等离子体传感器可以快速灵敏地检测 直接来自临床样品的疾病特异性EV。在这个项目中，我们将进一步推进技术， 强大的多重EV分析，并实现芯片上EV分离，以实现简单的分析程序， 重现性好。我们将使用成熟的临床前和临床样本验证该系统， 证明新技术在临床应用上的可行性和潜力。成功完成 该项目将产生一个高灵敏度的传感平台，用于多路复用EV分析。的发展 这种技术可以为理解亚型、异质性和生产提供更多的见解 在疾病发展和进展期间的EV动力学。获得的见解将铺平道路， 将EV研究扩展到各种疾病，进一步拓宽EV在临床环境中的应用范围。

Extracellular vesicles (EVs) present new opportunities for molecular diagnostics from non-invasive liquid biopsies. These cell-derived membrane-bound vesicles are abundantly present in biological fluids. EVs carry cell-specific cargos (e.g., lipids, proteins, and genetic materials), which can be harnessed to probe the molecular status of their cellular origins. EV analyses, however, pose unique technical challenges due to EVs' nanometer-sizes and presence in a vast biological background. EV analyses, however, pose unique technical challenges due to EVs' nanometer-sizes and presence in a vast biological background. While new technologies for EV analysis have been developed, fundamental limitations still remain, including i) low sensitivity limited to bulk analyses; ii) necessities of EV lysis for detecting markers inside of EVs; iii) lack of multiplexed analysis on protein and RNA markers; and iv) a separate EV isolation process required prior to the assay. The overall goal of this application is to overcome these technical challenges and develop a new platform that enables multiplexed analyses of EV protein and RNA markers in individual EVs. We previously developed a nanoplasmonic EV sensing platform based on transmission surface plasmon resonance through periodic nanohole gratings. We showed that the nanoplasmonic sensors could rapidly and sensitively detect disease-specific EVs directly from clinical samples. In this project, we will further advance the technology for robust multiplexed EV analysis and implement on-chip EV isolation to achieve simple assay procedures and good reproducibility. We will validate the system using well-established preclinical and clinical samples to demonstrate the feasibility and potential of the new technology for clinical applications. Successful completion of the project will produce a highly sensitive sensing platform for multiplexed EV analysis. The development of such a technology could offer additional insight into understanding subtypes, heterogeneity, and production dynamics of EVs during disease development and progression. The gained insights will pave the way for expanding EV studies to various diseases, further broadening the scope of EV applications in clinical settings.