The importance of extracellular vesicle purification for downstream analysis: A comparison of differential centrifugation and size exclusion chromatography for helminth pathogens

Chelsea N.  Davis; Helen  Phillips; John J.  Tomes; Martin T.  Swain; Toby Wilkinson; Peter Brophy; Russell M Morphew

doi:10.1371/journal.pntd.0007191

The importance of extracellular vesicle purification for downstream analysis: A comparison of differential centrifugation and size exclusion chromatography for helminth pathogens

Chelsea N. Davis, Helen Phillips, John J. Tomes, Martin T. Swain, Toby Wilkinson, Peter Brophy, Russell M Morphew

Research output: Contribution to journal › Article › peer-review

Abstract

Robust protocols for the isolation of extracellular vesicles (EVs) from the rest of their excretory-
secretory products are necessary for downstream studies and application development.
The most widely used purification method of EVs for helminth pathogens is currently
differential centrifugation (DC). In contrast, size exclusion chromatography (SEC) has been
included in the purification pipeline for EVs from other pathogens, highlighting there is not an
agreed research community ‘gold standard’ for EV isolation. In this case study, Fasciola
hepatica from natural populations were cultured in order to collect EVs from culture media
and evaluate a SEC or DC approach to pathogen helminth EV purification

Original language	English
Journal	PLoS Neglected Tropical Diseases
DOIs	https://doi.org/10.1371/journal.pntd.0007191
Publication status	Published - 27 Feb 2019

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abstract = "Robust protocols for the isolation of extracellular vesicles (EVs) from the rest of their excretory- secretory products are necessary for downstream studies and application development. The most widely used purification method of EVs for helminth pathogens is currently differential centrifugation (DC). In contrast, size exclusion chromatography (SEC) has been included in the purification pipeline for EVs from other pathogens, highlighting there is not an agreed research community {\textquoteleft}gold standard{\textquoteright} for EV isolation. In this case study, Fasciola hepatica from natural populations were cultured in order to collect EVs from culture media and evaluate a SEC or DC approach to pathogen helminth EV purification",

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AU - Wilkinson, Toby

AU - Brophy, Peter

AU - Morphew, Russell M

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AB - Robust protocols for the isolation of extracellular vesicles (EVs) from the rest of their excretory- secretory products are necessary for downstream studies and application development. The most widely used purification method of EVs for helminth pathogens is currently differential centrifugation (DC). In contrast, size exclusion chromatography (SEC) has been included in the purification pipeline for EVs from other pathogens, highlighting there is not an agreed research community ‘gold standard’ for EV isolation. In this case study, Fasciola hepatica from natural populations were cultured in order to collect EVs from culture media and evaluate a SEC or DC approach to pathogen helminth EV purification

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The importance of extracellular vesicle purification for downstream analysis: A comparison of differential centrifugation and size exclusion chromatography for helminth pathogens

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