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 μL 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.

中文翻译：

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纳米胞囊内系统分析囊泡内外泌体蛋白

胞外小泡（EVs），包括外泌体，是从细胞中脱落的纳米级膜颗粒，并含有细胞蛋白，其组成可能有助于癌症的诊断和治疗。大多数分析都集中在表面蛋白上，而囊内蛋白的分析则更具挑战性。在本文中，我们报告了使用纳米等离激元传感器对囊内和跨膜蛋白进行的EV筛选测定。该平台被称为iNPS（囊内纳米等离子体系统），使用基于纳米孔的表面等离子体共振（SPR）进行分子检测。具体来说，我们（i）建立了统一的检测方案以检测囊泡内和跨膜蛋白；（ii）工程化的等离激元底物，以提高检测灵敏度。生成的iNPS启用了敏感功能（0。每个标记5μL样品）和高通量（10×10阵列）检测EV蛋白。当用于监测来自药物治疗的癌细胞的EV时，iNPS分析揭示了药物依赖性的独特EV蛋白特征。我们设想，iNPS可能是电动汽车全面分子筛查的强大工具。