Park Jongmin, Im Hyungsoon, Hong Seonki, Castro Cesar M, Weissleder Ralph, Lee Hakho
Center for Systems Biology, Massachusetts General Hospital, 185 Cambridge St, CPZN 5206, Boston, MA 02114.
Department of Radiology, Massachusetts General Hospital, Boston, MA 02114.
ACS Photonics. 2018 Feb 21;5(2):487-494. doi: 10.1021/acsphotonics.7b00992. Epub 2017 Nov 3.
Extracellular vesicles (EVs), including exosomes, are nanoscale membrane particles shed from cells and contain cellular proteins whose makeup could inform cancer diagnosis and treatment. Most analyses have focused on surface proteins while analysis of intravesicular proteins has been more challenging. Herein, we report an EV screening assay for both intravesicular and transmembrane proteins using a nanoplasmonic sensor. Termed iNPS (intravesicular nanoplasmonic system), this platform used nanohole-based surface plasmon resonance (SPR) for molecular detection. Specifically, we i) established a unified assay protocol to detect intravesicular as well as transmembrane proteins; and ii) engineered plasmonic substrates to enhance detection sensitivity. The resulting iNPS enabled sensitive (0.5 L sample per marker) and high-throughput (a 10 × 10 array) detection for EV proteins. When applied to monitor EVs from drug-treated cancer cells, the iNPS assay revealed drug-dependent unique EV protein signatures. We envision that iNPS could be a powerful tool for comprehensive molecular screening of EVs.
细胞外囊泡（EVs），包括外泌体，是从细胞脱落的纳米级膜颗粒，含有细胞蛋白质，其组成可用于癌症诊断和治疗。大多数分析都集中在表面蛋白上，而对囊泡内蛋白的分析则更具挑战性。在此，我们报告了一种使用纳米等离子体传感器对囊泡内蛋白和跨膜蛋白进行EV筛选的检测方法。这个名为iNPS（囊泡内纳米等离子体系统）的平台利用基于纳米孔的表面等离子体共振（SPR）进行分子检测。具体而言，我们：i）建立了一个统一的检测方案来检测囊泡内蛋白和跨膜蛋白；ii）设计了等离子体基底以提高检测灵敏度。由此产生的iNPS能够对EV蛋白进行灵敏（每个标记物0.5微升样品）和高通量（10×10阵列）检测。当应用于监测来自药物处理癌细胞的EVs时，iNPS检测揭示了与药物相关的独特EV蛋白特征。我们设想iNPS可能成为一种用于EVs全面分子筛选的强大工具。
