, 1 February 2026, 139013
Multiplexed phenotypic profiling of single extracellular vesicles for early clinical diagnosis of high-grade serious ovarian carcinoma
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- Key Laboratory of Haikou Trauma, Key Laboratory of Hainan Trauma and Disaster Rescue, Key Laboratory of Emergency and Trauma, Ministry of Education, The First Affiliated Hospital of Hainan Medical University, Hainan Medical University, Haikou 571199, China
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- Engineering Research Center for Hainan Bio-Smart Materials and Bio-Medical Devices, College of Emergency and Trauma, The Second Affiliated Hospital of Hainan Medical University, Hainan Cancer Hospital, Hainan Medical University, Haikou 571199, China
Received 28 June 2025, Revised 13 October 2025, Accepted 20 October 2025, Available online 23 October 2025, Version of Record 28 October 2025.
Highlights
?A new approach to detect multiple phenotype changes of individual EVs on a fluorescence-microfluidic platform.
?A new assay for early-stage clinical diagnosis of HGSOC with high accuracy.
?Parallel microchambers on microchip enable high throughput detection of single EVs.
?Developed fluorescence-microchip approach outperformed traditional ELISA.
Abstract
Liquid biopsy based early detection of cancer is full of challenges mainly attributing to the lack of specific biomarkers and effective techniques. Extracellular vesicles (EVs) are demonstrated to be emerging biomarkers for early cancer detection. Due to their nanoscale size and heterogeneity induced by distinct biogenesis, the analysis of EVs in single vesicle level is still challenging. Here we develop a single EV detection assay on a fluorescence-microfluidic chip (SEAFM) to probe phenotype changes of single EVs for early detection of cancer, with speci?c focus on high-grade serous ovarian carcinoma (HGSOC) which has the highest mortality rate among malignant tumors in gynecology. The designed microfluidic chip possesses a parallel micro-chamber design, which is applied to high throughput EVs’ immobilization and detection. Four distinct fluorescently labelled antibodies are introduced into the platform for multiplexed detection of different EV surface biomarkers including ubiquitous EV biomarkers (i.e., tetraspanins; CD81) and putative HGSOC tumor biomarkers (i.e. EpCAM, CD24, EGFR). The phenotypic profiling of EVs is achieved by counting of fluorescent vesicles at a single particle level, which provides evidence for discriminating HGSOC patients from healthy controls. In addition, the developed approach surpassed conventional ELISA method in sensitivity, which has potential in early clinical diagnosis of HGSOC for improved personal treatment.
Keywords
Single extracellular vesicles
Microfluidics
Fluorescence labeling
Clinical diagnosis
Ovarian carcinoma
