Filter-aided extracellular vesicle enrichment (FAEVEr) for proteomics Article Swipe
Jarne Pauwels
,
Tessa Van de Steene
,
J. Van De Velde
,
Freya De Muyer
,
Dirk De Pauw
,
Femke Baeke
,
Sven Eyckerman
,
Kris Gevaert
·
YOU?
·
· 2023
· Open Access
·
· DOI: https://doi.org/10.1101/2023.07.06.547926
YOU?
·
· 2023
· Open Access
·
· DOI: https://doi.org/10.1101/2023.07.06.547926
Extracellular vesicles (EVs), membrane-delimited nanovesicles that are secreted by cells into the extracellular environment, are gaining substantial interest due to their involvement in cellular homeostasis and their contribution to disease pathology. The latter in particular has led to an exponential increase in interest in EVs as they are considered to be circulating packages containing potential biomarkers and are also a possible biological means to deliver drugs in a cell-specific manner. However, several challenges hamper straightforward proteome analysis of EVs as they are generally low abundant and reside in complex biological matrices. These matrices typically contain abundant protein concentrations that vastly exceed those of the EV proteome. Therefore, extensive EV isolation and purification protocols are imperative and many have been developed, including (density) ultracentrifugation, size-exclusion and precipitation methods. Here, we describe filter-aided extracellular vesicle enrichment (FAEVEr) as an approach based on 300 kDa MWCO filtration that allows the processing of multiple samples in parallel within a reasonable timeframe and at moderate cost. We demonstrate that FAEVEr is capable of quantitatively retaining EV particles on filters, whilst allowing extensive washing with the mild detergent TWEEN-20 to remove interfering non-EV proteins. The retained particles are directly lysed on the filter for a complete recovery of the EV protein cargo towards proteome analysis. Here, we validate and optimize FAEVEr on recombinant EV material and apply it on conditioned medium as well as on complex serum. Our results indicate that EVs isolated from MCF7 cells cultured with or without serum have a drastic different proteome because of nutrient deprivation.
Related Topics
Concepts
Proteome
Extracellular vesicles
Extracellular vesicle
Lysis
Extracellular
Chemistry
Proteomics
Ultracentrifuge
Microvesicles
Cell biology
Recombinant DNA
Vesicle
Biophysics
Biology
Membrane
Chromatography
Biochemistry
Gene
microRNA
Metadata
- Type
- preprint
- Language
- en
- Landing Page
- https://doi.org/10.1101/2023.07.06.547926
- https://www.biorxiv.org/content/biorxiv/early/2023/07/07/2023.07.06.547926.full.pdf
- OA Status
- green
- Cited By
- 1
- References
- 83
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4383499251
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
-
https://openalex.org/W4383499251Canonical identifier for this work in OpenAlex
- DOI
-
https://doi.org/10.1101/2023.07.06.547926Digital Object Identifier
- Title
-
Filter-aided extracellular vesicle enrichment (FAEVEr) for proteomicsWork title
- Type
-
preprintOpenAlex work type
- Language
-
enPrimary language
- Publication year
-
2023Year of publication
- Publication date
-
2023-07-07Full publication date if available
- Authors
-
Jarne Pauwels, Tessa Van de Steene, J. Van De Velde, Freya De Muyer, Dirk De Pauw, Femke Baeke, Sven Eyckerman, Kris GevaertList of authors in order
- Landing page
-
https://doi.org/10.1101/2023.07.06.547926Publisher landing page
- PDF URL
-
https://www.biorxiv.org/content/biorxiv/early/2023/07/07/2023.07.06.547926.full.pdfDirect link to full text PDF
- Open access
-
YesWhether a free full text is available
- OA status
-
greenOpen access status per OpenAlex
- OA URL
-
https://www.biorxiv.org/content/biorxiv/early/2023/07/07/2023.07.06.547926.full.pdfDirect OA link when available
- Concepts
-
Proteome, Extracellular vesicles, Extracellular vesicle, Lysis, Extracellular, Chemistry, Proteomics, Ultracentrifuge, Microvesicles, Cell biology, Recombinant DNA, Vesicle, Biophysics, Biology, Membrane, Chromatography, Biochemistry, Gene, microRNATop concepts (fields/topics) attached by OpenAlex
- Cited by
-
1Total citation count in OpenAlex
- Citations by year (recent)
-
2025: 1Per-year citation counts (last 5 years)
- References (count)
-
83Number of works referenced by this work
- Related works (count)
-
10Other works algorithmically related by OpenAlex
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