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Pinched-flow fractionation-based extracellular vesicle isolation by ExoFAST and its analytical benchmarking with
Chaithanya Chelakkot1, Yanymee N Guillen-Quispe1,2, Jongjin Jo3
1Research Institute of Pharmaceutical Science, Department of Pharmacy, Seoul National University College of Pharmacy, Seoul, Republic of Korea. lakshmi@snu.ac.kr.
The Analyst
|March 6, 2026
Summary
This study introduces ExoFAST, an automated microfluidic device for isolating extracellular vesicles (EVs). ExoFAST offers reproducible, operator-independent EV isolation, crucial for developing non-invasive liquid biopsy biomarkers.
Area of Science:
- Biotechnology
- Nanotechnology
- Biomedical Engineering
Background:
- Extracellular vesicles (EVs), especially exosomes, are key liquid biopsy biomarkers for non-invasive diagnostics.
- Challenges in robust and reproducible EV isolation hinder clinical translation of EV-based biomarkers.
Purpose of the Study:
- To evaluate an automated microfluidic workflow (ExoFAST) for EV isolation.
- To benchmark ExoFAST performance against established commercial methods.
Main Methods:
- ExoFAST utilizes pinched-flow fractionation for size-selective separation of small EVs.
- Analytical performance was assessed using standardized metrics (concentration, yield, size, marker expression).
- Benchmarking included ultracentrifugation, size-exclusion chromatography, polymer precipitation, and membrane filtration.
Main Results:
- ExoFAST achieved particle recovery comparable to ultracentrifugation with similar particle concentrations.
- The device demonstrated reproducible, operator-independent processing.
- ExoFAST successfully isolated EVs from various biological samples, including plasma and serum.
Conclusions:
- ExoFAST is a validated, automated, size-based EV enrichment workflow.
- Its controlled fractionation and transparent metrics support integration into EV characterization pipelines.
- This microfluidic approach addresses critical needs for clinical translation of EV biomarkers.
