Automated disc device for multiplexed extracellular vesicle isolation and labelling from liquid biopsies in cancer diagnostics

Abstract

Circulating extracellular vesicles can be used for tumour diagnostics. However, current isolation methods are time consuming, require manual handling and are prone to contamination. Here we report on SpinEx (separation-processing integration for extracellular vesicles), a compact disc device for automatic isolation and multiplex immunolabelling of whole-blood samples. SpinEx integrates on-disc chromatography, centripetal liquid transfer and bead-based vesicle capture with antibody labelling. The system processes 150 mu l of whole blood, enriching and labelling vesicles for 16 protein targets in under 75 minutes. Detection is performed by measuring dual fluorescence signals from labelled extracellular vesicles captured on microbeads. In a pilot clinical study, SpinEx was used to process 221 plasma samples for multiplex profiling of 30 vesicle-associated proteins. Using fluorescence flow cytometry to analyse cancer-specific biomarker expression, we found that vesicles processed by SpinEx distinguished cancer from non-cancer samples with 90% accuracy and 97% specificity, and classified 5 tumour types with 96% accuracy. SpinEx enables automated and multiplex processing of extracellular vesicles from blood, which may support the development of clinically viable assays for cancer detection and classification.