Related Experiment Video
Updated: Aug 23, 2025
09:16
Rapid Fluorescence-based Characterization of Single Extracellular Vesicles in Human Blood with Nanoparticle-tracking Analysis
Published on: January 7, 2019
9.9K
Characterizing single extracellular vesicles by droplet barcode sequencing for protein analysis.
Mahsan Banijamali1, Pontus Höjer1, Abel Nagy2
1Royal Institute of Technology (KTH), School of Engineering Sciences in Chemistry, Biotechnology and Health, Department of Gene Technology, Science for Life Laboratory, Solna, Sweden.
Journal of Extracellular Vesicles
|November 4, 2022
Summary
Droplet barcode sequencing for protein analysis (DBS-Pro) profiles individual small extracellular vesicles (sEVs), revealing sEV subtypes. This method enables precise surface protein analysis for disease biomarker discovery.
Area of Science:
- Biochemistry
- Molecular Biology
- Biotechnology
Background:
- Small extracellular vesicles (sEVs) are crucial biomarkers for disease diagnosis and monitoring.
- Current ensemble measurement techniques cannot resolve the heterogeneity of sEVs.
- Identifying sEV subtypes is essential for understanding their cellular origin.
Purpose of the Study:
- To introduce droplet barcode sequencing for protein analysis (DBS-Pro) for high-throughput single sEV profiling.
- To enable the identification and characterization of distinct sEV subtypes.
- To analyze surface proteins on individual sEVs from NSCLC cell lines and patient samples.
Main Methods:
- Compartmentalization of individual sEVs in emulsion droplets.
- Utilizing DNA-barcoded affinity reagents for multiplexed protein detection.
- Employing PCR-based droplet barcoding and sequencing for readout.
- Analysis of over 120,000 individual sEVs.
Main Results:
- DBS-Pro successfully profiles surface proteins on individual sEVs with a low mixing rate (<2%).
- The method facilitates the identification of sEV subtypes within heterogeneous samples.
- Demonstrated capability to analyze sEVs from both cell lines and clinical samples (NSCLC MPE fluid).
Conclusions:
- DBS-Pro is a powerful tool for single sEV proteomic profiling.
- This technology advances the characterization of sEV heterogeneity and subtype identification.
- Enables precise analysis of sEVs for improved disease diagnostics and therapeutic strategies.
