小体外囊泡（sEVs）携带着从母细胞继承而来的多种与肿瘤相关的蛋白质，在非侵袭性乳腺癌（BC）诊断中起到关键作用。然而，由于高度异质性的BC，评估sEV膜表面蛋白质的微小变异是具有挑战性的。因此，基于镧系（Ln3+）激活的荧光信号放大机制，我们开发了一种简单而超灵敏的检测方法，能够检测到BC来源的sEV上的多个表面蛋白质。通过NaEuF4基纳米探针的溶解放大荧光信号，能够以高灵敏度和特异性很好地识别sEVs上的多个蛋白生物标志物。我们运用线性判别分析成功区分三阴性BC细胞（MDA-MB-231细胞）来源的sEVs与其他乳腺细胞系（MCF-7、SK-BR-3、BT474和MCF-10A细胞）。此外，该策略还能够高准确度地区分BC患者和健康供体的进展阶段。这种简单而敏感的信号放大策略在通过对sEVs进行精确蛋白质分析方面展现了巨大的潜力，可用于早期临床诊断。版权所有©2023 Elsevier B.V.保留所有权利。

Small extracellular vesicles (sEVs) carrying multiple tumor-associated proteins inherited from parental cells play crucial roles in noninvasive breast cancer (BC) diagnosis. However, it is challenging to assess the subtle variations of surface proteins on sEV membranes due to the highly heterogeneous BC. Therefore, a simple and ultrasensitive assay based on lanthanide (Ln3+)-activated luminescence signal amplification was developed to detect multiple surface proteins on BC-derived sEVs. Multiple protein biomarkers on sEVs can be well identified with high sensitivity and specificity through dissolution-amplified luminescence of the NaEuF4 nanoparticle-based nanoprobe. We employ linear discriminant analysis to successfully discriminate triple negative BC cell (MDA-MB-231 cell) derived sEVs from other breast cell lines (MCF-7, SK-BR-3, BT474 and MCF-10A cell). Furthermore, the strategy enables high accuracy for districting the progression stages of BC patients and healthy donors. The simple and sensitive signal amplification strategy exhibits great potential for early clinic diagnosis by precise protein profiling of sEVs.Copyright © 2023 Elsevier B.V. All rights reserved.