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Related Experiment Video

Updated: Jun 7, 2025

Rapid Fluorescence-based Characterization of Single Extracellular Vesicles in Human Blood with Nanoparticle-tracking Analysis
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High-throughput, multiplexed quantification, and sorting of single EVs at single-molecule level.

Juhwan Park, Michelle Feng, Jingbo Yang

    Biorxiv : the Preprint Server for Biology
    |November 18, 2024
    PubMed
    Summary

    We developed BDEVS, a high-throughput platform for ultra-sensitive profiling of individual extracellular vesicles (EVs) in plasma. This technology enables precise quantification and sorting of EVs, advancing cancer immunology research and diagnostics.

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    Area of Science:

    • Biotechnology
    • Nanotechnology
    • Cancer Immunology

    Background:

    • Extracellular vesicles (EVs) are crucial in intercellular communication and disease progression.
    • Current methods for EV profiling lack throughput, sensitivity, or multiplexing capabilities for complex clinical samples.

    Purpose of the Study:

    • To develop a novel platform for high-throughput, multiplexed, and ultra-sensitive profiling of individual EVs directly in plasma.
    • To overcome limitations of existing technologies in EV analysis, enabling better understanding of tumor-immune interactions.

    Main Methods:

    • Developed BDEVS (Agarose Bead-based Digital Single Molecule-Single EV Sorting) platform.
    • Integrated rolling circle amplification (RCA) of cleaved EV surface proteins within agarose droplets.
    • Utilized flow cytometry for readout, quantification, and sorting of EVs.

    Main Results:

    • Achieved single molecule sensitivity and moderate multiplexing (3-plex) at high throughput (thousands of EVs/minute).
    • Demonstrated a ~100x improvement in the limit of detection for EV subpopulations compared to gold standards.
    • Successfully profiled and sorted EVs for immune markers (PD-L1, CD155) and a melanoma marker (TYRP-1) in plasma.

    Conclusions:

    • BDEVS offers unprecedented resolution for analyzing tumor-immune interactions and detecting rare EV subpopulations.
    • The platform shows transformative potential for EV-based diagnostics and therapeutic monitoring in cancer immunology.
    • BDEVS enables precise quantification and sorting of EVs, crucial for understanding disease heterogeneity in clinical specimens.