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Updated: Jun 3, 2026
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Chemical Affinity-Based Isolation of Extracellular Vesicles from Biofluids for Proteomics and Phosphoproteomics Analysis
Published on: October 27, 2023
Ammonium sulfate provides an efficient method for isolating small extracellular vesicles from human biofluids.
Qimeng Li1, Zinan Guo2, Yu Li2
1Department of Central Laboratory, Xuanwu Hospital of Capital Medical University, Beijing, 100053, China.
Scientific Reports
|June 1, 2026
Summary
We developed a fast and cost-effective ammonium sulfate method to isolate small extracellular vesicles (EVs) from biofluids for neurodegenerative disease diagnosis. This optimized pipeline offers high efficiency and practicality for clinical use.
Area of Science:
- Biochemistry
- Cell Biology
- Neuroscience
Background:
- Small extracellular vesicles (EVs) are key biomarkers for neurodegenerative diseases like Alzheimer's and Parkinson's.
- Current isolation methods are time-consuming, costly, and yield low purity, hindering clinical translation.
- A robust and efficient isolation technique is crucial for developing liquid biopsy approaches.
Purpose of the Study:
- To develop and validate an optimized, cost-effective ammonium sulfate (AS)-based pipeline for small EVs isolation.
- To refine the AS concentration for plasma and demonstrate feasibility in saliva.
- To introduce a targeted purification step for CNS-derived EVs and validate the protocol in a clinical cohort.
Main Methods:
- Utilized an optimized ammonium sulfate (AS) precipitation method based on the ExoPRISM framework.
- Determined optimal AS concentration (2.66 M) for small EVs isolation from plasma and saliva.
- Incorporated a purification step using GLAST antibodies for CNS-derived EVs and validated using NTA, electron microscopy, and marker detection (CD63, CD9).
Main Results:
- The AS-based method isolates small EVs in approximately 90 minutes with simple centrifugation (12,000 xg).
- Achieved comparable efficacy to commercial kits in isolating small EVs from plasma and purifying astrocyte-derived EVs (ADEVs).
- Demonstrated preservation of small EVs integrity and functional equivalence for downstream applications.
Conclusions:
- The optimized AS-based pipeline offers a robust, efficient, and cost-effective solution for small EVs isolation.
- This method significantly enhances temporal efficiency and reduces costs, making it highly suitable for clinical implementation.
- The AS-based approach holds significant translational potential for the early and accurate diagnosis of neurodegenerative diseases using liquid biopsies.
