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Updated: Jun 25, 2026
09:57
Isolation and Analysis of Traceable and Functionalized Extracellular Vesicles from the Plasma and Solid Tissues
Published on: October 17, 2022
1.9K
High-Efficiency Capture and Proteomic Analysis of Plasma-Derived Extracellular Vesicles through Affinity Purification
Guiyuan Zhang1,2,3, Chengxiao Ma4, Le Ma5
1School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China.
Analytical Chemistry
|February 5, 2025
Summary
Researchers developed EVlent, a magnetic bead method for isolating extracellular vesicles (EVs) from small plasma volumes. This technique enhances proteomic analysis and identifies potential Parkinson
Area of Science:
- Biochemistry
- Proteomics
- Nanotechnology
Background:
- Plasma-derived extracellular vesicles (EVs) are valuable biomarker sources.
- Isolating EVs from small human plasma volumes for proteomic analysis is challenging due to contaminants like proteins and lipoproteins.
- Existing methods like magnetic bead immune-isolation are incompatible with nanoparticle tracking analysis (NTA) and proteomic analysis.
Purpose of the Study:
- To develop a rapid and efficient method for isolating EVs from small plasma volumes for proteomic analysis.
- To overcome contamination issues hindering downstream analyses.
- To enable compatibility with multiple characterization techniques including NTA and proteomic analysis.
Main Methods:
- Development of a functional affinity magnetic bead, EVlent (Extracellular Vesicles isoLated Efficiently, Naturally, and Totally).
- Optimization of magnetic bead and plasma quantities for EV isolation.
- Characterization of isolated EVs using Western blotting (WB), NTA, and transmission electron microscopy (TEM).
- Proteomic analysis of isolated EVs from varying plasma volumes (100 μL and 5 μL).
- Comparison of EVlent with ultracentrifugation regarding contaminant reduction.
Main Results:
- Successful isolation of EVs from plasma using EVlent, confirmed by WB, NTA, and TEM.
- Proteomic analysis identified over 2000 proteins and 15,000 peptides from 100 μL plasma, and nearly 1000 proteins from 5 μL plasma.
- EVlent significantly reduced plasma protein and lipoprotein contaminants compared to ultracentrifugation.
- Application to healthy and Parkinson's disease patient samples identified four potential diagnostic biomarkers.
Conclusions:
- EVlent provides a rapid, efficient, and contamination-reducing method for isolating EVs from small plasma volumes.
- This method enables comprehensive proteomic analysis and supports biomarker discovery.
- The identified biomarkers show promise for clinical diagnosis of Parkinson's disease.
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