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Polyethylene glycol improves current methods for circulating extracellular vesicle-derived DNA isolation
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Polyethylene glycol improves current methods for circulating extracellular vesicle-derived DNA isolation
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Journal Article
Polyethylene glycol improves current methods for circulating extracellular vesicle-derived DNA isolation
2019
Overview
Background
Extracellular vesicles (EVs) are small membrane-bound vesicles which play an important role in cell-to-cell communication. Their molecular cargo analysis is presented as a new source for biomarker detection, and it might provide an alternative to traditional solid biopsies. However, the most effective approach for EV isolation is not yet well established.
Results
Here, we study the efficiency of the most common EV isolation methods-ultracentrifugation, Polyethlyene glycol and two commercial kits, Exoquick
®
and PureExo
®
. We isolated circulating EVs from the bloodstream of healthy donors, characterized the size and yield of EVs and analyzed their protein profiles and concentration. Moreover, we have used for the first time Digital-PCR to identify and detect specific gDNA sequences, which has several implications for diagnostic and monitoring many types of diseases.
Conclusions
Our findings present Polyethylene glycol precipitation as the most feasible and less cost-consuming EV isolation technique.
Publisher
BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
Subject
/ Biomedical and Life Sciences
/ Biopsy
/ Cell-Free Nucleic Acids - genetics
/ Cell-Free Nucleic Acids - isolation & purification
/ DNA
/ ExoQuick
/ Extracellular Vesicles - metabolism
/ Glycols (Class of compounds)
/ Humans
/ Methods
/ Polyethylene Glycols - pharmacology
/ Polyols
/ Proteins
/ PureExo
/ Urine
