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

细胞外小泡为非侵袭性液体的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体液中。电动汽车搭载 细胞特有的货物(例如，脂类、蛋白质和遗传物质)，可以被利用来探测 它们细胞起源的分子状态。然而，电动汽车分析带来了独特的技术挑战，因为电动汽车 纳米大小和存在于广阔的生物背景中。然而，电动汽车分析提出了独特的技术 由于电动汽车的纳米尺寸和在广阔的生物背景中的存在而带来的挑战。虽然是新的 电动汽车分析技术已经开发，但基本限制仍然存在，包括：i)低 灵敏度仅限于批量分析；ii)EV裂解用于检测EVS内部标志物的必要性；III)缺乏 对蛋白质和RNA标记的多重分析；以及iv)在进行 化验。这个应用程序的总体目标是克服这些技术挑战，开发一种新的 该平台能够对单个电动汽车中的EV蛋白质和RNA标记进行多路分析。我们之前 开发了一种基于透射式表面等离子体共振的纳米等离子体电动汽车传感平台 周期性纳米孔栅。我们证明了纳米等离子体传感器可以快速而灵敏地检测到 直接从临床样本中提取疾病特异性EVS。在这个项目中，我们将进一步推进 强大的多路复用EV分析和实施片上EV隔离，以实现简单的检测程序和 重现性好。我们将使用成熟的临床前和临床样本来验证该系统 论证了新技术在临床应用的可行性和潜力。成功完成 该项目的实施将为多路电动汽车分析提供一个高度灵敏的传感平台。的发展。 这样的技术可以为理解子类型、异质性和生产提供额外的洞察力 EVS在疾病发展和进展过程中的动态。所获得的见解将为 将EV研究扩展到各种疾病，进一步拓宽了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.