Journal
PRACTICAL LABORATORY MEDICINE
Volume 26, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.plabm.2021.e00241
Keywords
Exosomes; Size-exclusion chromatography; Ultracentrifugation; Mass spectrometry; Human plasma
Categories
Funding
- Cultural Affairs and Missions Sector, Ministry of Higher education, Arab Republic of Egypt
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The study found that using the UC + SEC method can more effectively enrich proteins in exosomes, and coupling UC and SEC can reduce plasma protein contamination by 98.8% compared to using UC alone.
Objectives: Exosomes are small lipid bilayer vesicles that are defined by their endocytic origin and size range of 30-140 nm. They are constantly produced by different cell types, by both healthy and abnormal cells, and can be isolated from almost all body fluids. Little information exists in isolating exosomes from plasma due to the complexity of its content and the presence of contaminating plasma proteins. Design and methods: We carried-out liquid chromatography-mass spectrometry (LC-MS/MS) analyses of plasma-derived vesicles from 4 healthy donors obtained by 2 coupled methodologies: Ultracentrifugation (UC) coupled with size-exclusion chromatography (SEC) to isolate and subsequently enrich exosomes. We compared the proteins detected by UC alone and UC coupled with SEC. Results: In the coupled UC + SEC methodology we found 52.25% more proteins enriched in exosomes as CD9, Annexins, YWHAZ (14-3-3 family) and others, than by using UC alone. There is also a reduction of 98.8% of contaminating plasma proteins by coupling UC and SEC in comparison to using UC alone. Conclusions: We conclude that exosomes can be successfully isolated from plasma using a very simple combination of standard methods, which could largely improve the proteomics profiling of plasma exosomes.
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