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Rapid Fluorescence-based Characterization of Single Extracellular Vesicles in Human Blood with Nanoparticle-tracking Analysis
Published on: January 7, 2019
10.0K
Framework for rapid comparison of extracellular vesicle isolation methods.
Dmitry Ter-Ovanesyan1, Maia Norman1,2,3, Roey Lazarovits1
1Wyss Institute for Biologically Inspired Engineering, Boston, United States.
Elife
|November 16, 2021
Summary
This study introduces ultrasensitive single-molecule array (Simoa) assays for quantifying extracellular vesicles (EVs) using common protein markers. This method improves the assessment and isolation of EVs from biological fluids like plasma and CSF.
Area of Science:
- Biochemistry
- Cell Biology
- Biomarker Discovery
Background:
- Extracellular vesicles (EVs) are crucial biomarkers found in biofluids.
- Current methods struggle to accurately quantify EVs and distinguish them from lipoproteins and protein aggregates.
Purpose of the Study:
- To develop and validate ultrasensitive single-molecule array (Simoa) assays for quantifying EVs.
- To compare the purity and efficiency of different EV isolation techniques.
Main Methods:
- Utilized Simoa assays to measure tetraspanins (CD9, CD63, CD81) as EV markers.
- Quantified albumin to assess protein contamination.
- Evaluated ultracentrifugation, precipitation, and size exclusion chromatography (SEC) for EV isolation from plasma and CSF.
Main Results:
- Simoa assays provide sensitive quantification of EVs using CD9, CD63, and CD81.
- Demonstrated improved purity and efficiency of EV isolation methods, particularly SEC.
- Successfully differentiated EVs from protein contaminants like albumin.
Conclusions:
- Simoa assays offer a powerful tool for quantifying EV proteins in biofluids.
- The developed framework enables rapid comparison and optimization of EV isolation strategies.
- This approach enhances the reliability of EV-based biomarker research.
