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Updated: May 20, 2025
Protein Digestion, Ultrafiltration, and Size Exclusion Chromatography to Optimize the Isolation of Exosomes from Human Blood Plasma and Serum
Published on: April 13, 2018
Comparison of Methods for Isolating Exosomes from Plasma Subjects with Normal and High Fat Percentages
Jacqueline Noboa-Velástegui1,2, Juan Carlos León3, Jorge Castro4
1Doctorado en Ciencias Biomédicas, Secretaría Académica, Centro Universitario de Ciencias de la Salud, Universidad de Guadalajara, Calle Sierra Mojada No. 950, Colonia Independencia, Guadalajara C.P. 44340, Mexico.
Researchers compared three plasma exosome isolation methods. Differential centrifugation (DC) and commercial kits (CK) are recommended for protein and molecular analyses, respectively, for studying immunometabolic alterations.
Area of Science:
- Extracellular vesicle (EV) research
- Immunometabolism
- Biomarker discovery
Background:
- Adipose tissue is crucial for fat storage and produces extracellular vesicles (EVs).
- Exosomes, a type of EV, carry biological cargo reflecting immunometabolic status.
- Understanding exosome content requires effective isolation techniques.
Purpose of the Study:
- To compare three plasma exosome isolation methods: commercial kit (CK), size exclusion chromatography (SEC), and differential centrifugation (DC).
- To identify the most advantageous method for exosome isolation based on specific research needs.
- To investigate potential differences in exosome cargo between individuals with normal and high body fat percentages.
Main Methods:
- Plasma samples were collected from individuals with normal and high body fat percentages.
- Exosomes were isolated using three distinct methods: CK, SEC, and DC.
- Exosome quality, morphology, total protein, and microRNA concentration were analyzed.
Main Results:
- Differential centrifugation (DC) and commercial kits (CK) were identified as the most advantageous isolation methods.
- DC is recommended for protein analysis, while CK is suggested for molecular analysis.
- No significant statistical differences were found in exosome quality, morphology, total protein, or microRNA concentration based on body fat percentage.
Conclusions:
- The choice of exosome isolation method (CK, SEC, or DC) should align with specific research objectives.
- Despite no observed differences in overall exosome characteristics, the exosomal cargo composition likely varies between individuals with normal and high body fat percentages.
- Further research is warranted to elucidate the specific variations in exosomal cargo related to body fat percentage and immunometabolic alterations.
