Different isolation approaches lead to diverse glycosylated extracellular vesicle populations

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Standard

Different isolation approaches lead to diverse glycosylated extracellular vesicle populations. / Freitas, Daniela; Balmaña, Meritxell; Poças, Juliana; Campos, Diana; Osório, Hugo; Konstantinidi, Andriana; Vakhrushev, Sergey Y.; Magalhães, Ana; Reis, Celso A.

I: Journal of Extracellular Vesicles, Bind 8, Nr. 1, 1621131, 2019.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningfagfællebedømt

Harvard

Freitas, D, Balmaña, M, Poças, J, Campos, D, Osório, H, Konstantinidi, A, Vakhrushev, SY, Magalhães, A & Reis, CA 2019, 'Different isolation approaches lead to diverse glycosylated extracellular vesicle populations', Journal of Extracellular Vesicles, bind 8, nr. 1, 1621131. https://doi.org/10.1080/20013078.2019.1621131

APA

Freitas, D., Balmaña, M., Poças, J., Campos, D., Osório, H., Konstantinidi, A., ... Reis, C. A. (2019). Different isolation approaches lead to diverse glycosylated extracellular vesicle populations. Journal of Extracellular Vesicles, 8(1), [1621131]. https://doi.org/10.1080/20013078.2019.1621131

Vancouver

Freitas D, Balmaña M, Poças J, Campos D, Osório H, Konstantinidi A o.a. Different isolation approaches lead to diverse glycosylated extracellular vesicle populations. Journal of Extracellular Vesicles. 2019;8(1). 1621131. https://doi.org/10.1080/20013078.2019.1621131

Author

Freitas, Daniela ; Balmaña, Meritxell ; Poças, Juliana ; Campos, Diana ; Osório, Hugo ; Konstantinidi, Andriana ; Vakhrushev, Sergey Y. ; Magalhães, Ana ; Reis, Celso A. / Different isolation approaches lead to diverse glycosylated extracellular vesicle populations. I: Journal of Extracellular Vesicles. 2019 ; Bind 8, Nr. 1.

Bibtex

@article{b5fd7ef61f614f0fb77fae385fdaf574,
title = "Different isolation approaches lead to diverse glycosylated extracellular vesicle populations",
abstract = "Extracellular vesicles (EVs) are a heterogeneous group of small secreted particles involved in intercellular communication and mediating a broad spectrum of biological functions. EVs cargo is composed of a large repertoire of molecules, including glycoconjugates. Herein, we report the first study on the impact of the isolation strategy on the EV populations’ glycosylation profile. The use of different state-of-the-art protocols, namely differential ultracentrifugation (UC), total exosome isolation (TEI), OptiPrepTM density gradient (ODG) and size exclusion chromatography (SEC) resulted in EV populations displaying different sets of glycoconjugates. The EV populations obtained by UC, ODG and SEC methods displayed similar protein and glycan profiles, whereas TEI methodology isolated the most distinct EV population. In addition, ODG and SEC isolation protocols provided an enhanced EV glycoproteins detection. Remarkably, proteins displaying the tumour-associated glycan sialyl-Tn (STn) were identified as packaged cargo into EVs independently of the isolation methodology. STn carrying EV samples isolated by UC, ODG and SEC presented a considerable set of cancer-related proteins that were not detected in EVs isolated by TEI. Our work demonstrates the impact of using different isolation methodologies in the populations of EVs that are obtained, with consequences in the glycosylation profile of the isolated population. Furthermore, our results highlight the importance of selecting adequate EV isolation protocols and cell culture conditions to determine the structural and functional complexity of the EV glycoconjugates.",
keywords = "Extracellular vesicles, glycosylation, isolation protocols, OptiPrep density gradient, size exclusion chromatography, total exosome isolation, ultracentrifugation",
author = "Daniela Freitas and Meritxell Balma{\~n}a and Juliana Po{\cc}as and Diana Campos and Hugo Os{\'o}rio and Andriana Konstantinidi and Vakhrushev, {Sergey Y.} and Ana Magalh{\~a}es and Reis, {Celso A.}",
year = "2019",
doi = "10.1080/20013078.2019.1621131",
language = "English",
volume = "8",
journal = "Journal of Extracellular Vesicles",
issn = "2001-3078",
publisher = "Co-Action Publishing",
number = "1",

}

RIS

TY - JOUR

T1 - Different isolation approaches lead to diverse glycosylated extracellular vesicle populations

AU - Freitas, Daniela

AU - Balmaña, Meritxell

AU - Poças, Juliana

AU - Campos, Diana

AU - Osório, Hugo

AU - Konstantinidi, Andriana

AU - Vakhrushev, Sergey Y.

AU - Magalhães, Ana

AU - Reis, Celso A.

PY - 2019

Y1 - 2019

N2 - Extracellular vesicles (EVs) are a heterogeneous group of small secreted particles involved in intercellular communication and mediating a broad spectrum of biological functions. EVs cargo is composed of a large repertoire of molecules, including glycoconjugates. Herein, we report the first study on the impact of the isolation strategy on the EV populations’ glycosylation profile. The use of different state-of-the-art protocols, namely differential ultracentrifugation (UC), total exosome isolation (TEI), OptiPrepTM density gradient (ODG) and size exclusion chromatography (SEC) resulted in EV populations displaying different sets of glycoconjugates. The EV populations obtained by UC, ODG and SEC methods displayed similar protein and glycan profiles, whereas TEI methodology isolated the most distinct EV population. In addition, ODG and SEC isolation protocols provided an enhanced EV glycoproteins detection. Remarkably, proteins displaying the tumour-associated glycan sialyl-Tn (STn) were identified as packaged cargo into EVs independently of the isolation methodology. STn carrying EV samples isolated by UC, ODG and SEC presented a considerable set of cancer-related proteins that were not detected in EVs isolated by TEI. Our work demonstrates the impact of using different isolation methodologies in the populations of EVs that are obtained, with consequences in the glycosylation profile of the isolated population. Furthermore, our results highlight the importance of selecting adequate EV isolation protocols and cell culture conditions to determine the structural and functional complexity of the EV glycoconjugates.

AB - Extracellular vesicles (EVs) are a heterogeneous group of small secreted particles involved in intercellular communication and mediating a broad spectrum of biological functions. EVs cargo is composed of a large repertoire of molecules, including glycoconjugates. Herein, we report the first study on the impact of the isolation strategy on the EV populations’ glycosylation profile. The use of different state-of-the-art protocols, namely differential ultracentrifugation (UC), total exosome isolation (TEI), OptiPrepTM density gradient (ODG) and size exclusion chromatography (SEC) resulted in EV populations displaying different sets of glycoconjugates. The EV populations obtained by UC, ODG and SEC methods displayed similar protein and glycan profiles, whereas TEI methodology isolated the most distinct EV population. In addition, ODG and SEC isolation protocols provided an enhanced EV glycoproteins detection. Remarkably, proteins displaying the tumour-associated glycan sialyl-Tn (STn) were identified as packaged cargo into EVs independently of the isolation methodology. STn carrying EV samples isolated by UC, ODG and SEC presented a considerable set of cancer-related proteins that were not detected in EVs isolated by TEI. Our work demonstrates the impact of using different isolation methodologies in the populations of EVs that are obtained, with consequences in the glycosylation profile of the isolated population. Furthermore, our results highlight the importance of selecting adequate EV isolation protocols and cell culture conditions to determine the structural and functional complexity of the EV glycoconjugates.

KW - Extracellular vesicles

KW - glycosylation

KW - isolation protocols

KW - OptiPrep density gradient

KW - size exclusion chromatography

KW - total exosome isolation

KW - ultracentrifugation

U2 - 10.1080/20013078.2019.1621131

DO - 10.1080/20013078.2019.1621131

M3 - Journal article

C2 - 31236201

AN - SCOPUS:85074725417

VL - 8

JO - Journal of Extracellular Vesicles

JF - Journal of Extracellular Vesicles

SN - 2001-3078

IS - 1

M1 - 1621131

ER -

ID: 235593443