A new isolation method for bacterial extracellular vesicles providing greater purity and improved proteomic detection of vesicle proteins.

Le, LHM; Steele, JR; Ying, L; Schittenhelm, RB; Ferrero, RL

A new isolation method for bacterial extracellular vesicles providing greater purity and improved proteomic detection of vesicle proteins.

Le, LHM Steele, JR Ying, L Schittenhelm, RB Ferrero, RL

Permalink

Publisher:: Wiley
Publication Type:: Journal Article
Citation:: J Extracell Biol, 2023, 2, (5), pp. e84
Issue Date:: 2023-05

Recently Added

	Filename	Description	Size
	J of Extracellular Bio - 2023 - Le - A new isolation method for bacterial extracellular vesicles providing greater purity.pdf	Published version	4.11 MB	Adobe PDF	View/Open

Copyright Clearance Process

Recently Added
In Progress
Open Access

This item is new to OPUS and is not currently available.

Full metadata record

Field	Value	Language
dc.contributor.author	Le, LHM
dc.contributor.author	Steele, JR
dc.contributor.author	Ying, L
dc.contributor.author	Schittenhelm, RB
dc.contributor.author	Ferrero, RL
dc.date.accessioned	2024-07-22T01:19:47Z
dc.date.available	2023-04-02
dc.date.available	2024-07-22T01:19:47Z
dc.date.issued	2023-05
dc.identifier.citation	J Extracell Biol, 2023, 2, (5), pp. e84
dc.identifier.issn	2768-2811
dc.identifier.issn	2768-2811
dc.identifier.uri	http://hdl.handle.net/10453/179805
dc.description.abstract	Contaminants within cell culture media often co-isolate with eukaryotic extracellular vesicles (EVs) thus affecting their biological properties. It has yet to be investigated if this is also true for bacterial EVs (BEVs), especially for organisms grown in complex culture media containing animal-derived products. To address this question, we isolated BEVs from the fastidious bacterium Helicobacter pylori grown in either standard Brain Heart Infusion (BHI) medium or BHI depleted of animal-derived products (D-BHI). We show that BEVs prepared from bacteria grown in D-BHI medium have similar morphologies, size ranges and yields to those prepared from standard medium. Similarly, no differences were found in the ability of H. pylori BEVs to induce IL-8 responses in epithelial cells. However, H. pylori BEVs prepared from D-BHI medium were of higher purity than those prepared from standard medium. Importantly, proteomic analyses detected 3.4-fold more H. pylori proteins and 10-fold fewer bovine-derived proteins in BEV samples prepared from D-BHI rather than the standard method. Fifty-seven H. pylori proteins were uniquely detected in BEV samples prepared from D-BHI. In conclusion, we have described an improved method for BEV isolation. Furthermore, we demonstrate how animal-derived products in bacteriological culture media may adversely affect proteomic analyses of BEVs.
dc.format	Electronic-eCollection
dc.language	eng
dc.publisher	Wiley
dc.relation.ispartof	J Extracell Biol
dc.relation.isbasedon	10.1002/jex2.84
dc.rights	info:eu-repo/semantics/restrictedAccess
dc.title	A new isolation method for bacterial extracellular vesicles providing greater purity and improved proteomic detection of vesicle proteins.
dc.type	Journal Article
utslib.citation.volume	2
utslib.location.activity	United States
pubs.organisational-group	University of Technology Sydney
pubs.organisational-group	University of Technology Sydney/Provost
pubs.organisational-group	University of Technology Sydney/Provost/Jumbunna
utslib.copyright.status	recently_added	*
dc.date.updated	2024-07-22T01:19:43Z
pubs.issue	5
pubs.publication-status	Published online
pubs.volume	2
utslib.citation.issue	5

Abstract:

Contaminants within cell culture media often co-isolate with eukaryotic extracellular vesicles (EVs) thus affecting their biological properties. It has yet to be investigated if this is also true for bacterial EVs (BEVs), especially for organisms grown in complex culture media containing animal-derived products. To address this question, we isolated BEVs from the fastidious bacterium Helicobacter pylori grown in either standard Brain Heart Infusion (BHI) medium or BHI depleted of animal-derived products (D-BHI). We show that BEVs prepared from bacteria grown in D-BHI medium have similar morphologies, size ranges and yields to those prepared from standard medium. Similarly, no differences were found in the ability of H. pylori BEVs to induce IL-8 responses in epithelial cells. However, H. pylori BEVs prepared from D-BHI medium were of higher purity than those prepared from standard medium. Importantly, proteomic analyses detected 3.4-fold more H. pylori proteins and 10-fold fewer bovine-derived proteins in BEV samples prepared from D-BHI rather than the standard method. Fifty-seven H. pylori proteins were uniquely detected in BEV samples prepared from D-BHI. In conclusion, we have described an improved method for BEV isolation. Furthermore, we demonstrate how animal-derived products in bacteriological culture media may adversely affect proteomic analyses of BEVs.

Please use this identifier to cite or link to this item:

http://hdl.handle.net/10453/179805