A simple immunoassay for extracellular vesicle liquid biopsy in microliters of non-processed plasma

[Background]: Extracellular vesicles (EVs), released by most cell types, provide an excellent source of biomarkers in biological fluids. However, in order to perform validation studies and screenings of patient samples, it is still necessary to develop general techniques permitting rapid handling of...

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Authors: Campos-Silva, Carmen, Cáceres-Martell, Yaiza, Sánchez-Herrero, Estela, Sandúa, Amaia, Beneítez-Martínez, Alexandra, González, Álvaro, Provencio, Mariano, Romero, Atocha, Jara-Acevedo, Ricardo, Yáñez-Mó, María, Valés-Gómez, Mar
Format: article
Status:Published version
Publication Date:2022
Country:España
Institution:Consejo Superior de Investigaciones Científicas (CSIC)
Repository:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/346346
Online Access:http://hdl.handle.net/10261/346346
Access Level:Open access
Keyword:Extracellular vesicles
Cancer
Colloids
Flocculation
ELISA
Flow cytometry
Liquid biopsy
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spelling A simple immunoassay for extracellular vesicle liquid biopsy in microliters of non-processed plasmaCampos-Silva, CarmenCáceres-Martell, YaizaSánchez-Herrero, EstelaSandúa, AmaiaBeneítez-Martínez, AlexandraGonzález, ÁlvaroProvencio, MarianoRomero, AtochaJara-Acevedo, RicardoYáñez-Mó, MaríaValés-Gómez, MarExtracellular vesiclesCancerColloidsFlocculationELISAFlow cytometryLiquid biopsy[Background]: Extracellular vesicles (EVs), released by most cell types, provide an excellent source of biomarkers in biological fluids. However, in order to perform validation studies and screenings of patient samples, it is still necessary to develop general techniques permitting rapid handling of small amounts of biological samples from large numbers of donors.[Results]: Here we describe a method that, using just a few microliters of patient’s plasma, identifies tumour markers exposed on EVs. Studying physico-chemical properties of EVs in solution, we demonstrate that they behave as stable colloidal suspensions and therefore, in immunocapture assays, many of them are unable to interact with a stationary functionalised surface. Using flocculation methods, like those used to destabilize colloids, we demonstrate that cationic polymers increase EV ζ-potential, diameter, and sedimentation coefficient and thus, allow a more efficient capture on antibody-coated surfaces by both ELISA and bead-assisted flow cytometry. These findings led to optimization of a protocol in microtiter plates allowing effective immunocapture of EVs, directly in plasma without previous ultracentrifugation or other EV enrichment. The method, easily adaptable to any laboratory, has been validated using plasma from lung cancer patients in which the epithelial cell marker EpCAM has been detected on EVs.[Conclusions]: This optimized high throughput, easy to automate, technology allows screening of large numbers of patients to phenotype tumour markers in circulating EVs, breaking barriers for the validation of proposed EV biomarkers and the discovery of new ones.This work was supported by the Spanish Ministry of Science and Innovation (MCIU/AEI/FEDER, EU) under Grants RTI2018-093569-B-I00 and RED2018-102411-T (Translational Network for clinical application of EV, Tentacles); Madrid Regional Government under Grant “IMMUNOTHERCAN” (S2017/BMD-3733-2) and IND2019/BMD-17258.Peer reviewedScientific Research PublishingMinisterio de Ciencia, Innovación y Universidades (España)Agencia Estatal de Investigación (España)Comunidad de MadridConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202420242022info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/346346reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/RTI2018-093569-B-I00info:eu-repo/grantAgreement/AEI//RED2018-102411-TS2017/BMD-3733-2/IMMUNOTHERCANIND2019/BMD-17258https://doi.org/10.1186/s12951-022-01256-5Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/3463462026-05-22T06:33:51Z
dc.title.none.fl_str_mv A simple immunoassay for extracellular vesicle liquid biopsy in microliters of non-processed plasma
title A simple immunoassay for extracellular vesicle liquid biopsy in microliters of non-processed plasma
spellingShingle A simple immunoassay for extracellular vesicle liquid biopsy in microliters of non-processed plasma
Campos-Silva, Carmen
Extracellular vesicles
Cancer
Colloids
Flocculation
ELISA
Flow cytometry
Liquid biopsy
title_short A simple immunoassay for extracellular vesicle liquid biopsy in microliters of non-processed plasma
title_full A simple immunoassay for extracellular vesicle liquid biopsy in microliters of non-processed plasma
title_fullStr A simple immunoassay for extracellular vesicle liquid biopsy in microliters of non-processed plasma
title_full_unstemmed A simple immunoassay for extracellular vesicle liquid biopsy in microliters of non-processed plasma
title_sort A simple immunoassay for extracellular vesicle liquid biopsy in microliters of non-processed plasma
dc.creator.none.fl_str_mv Campos-Silva, Carmen
Cáceres-Martell, Yaiza
Sánchez-Herrero, Estela
Sandúa, Amaia
Beneítez-Martínez, Alexandra
González, Álvaro
Provencio, Mariano
Romero, Atocha
Jara-Acevedo, Ricardo
Yáñez-Mó, María
Valés-Gómez, Mar
author Campos-Silva, Carmen
author_facet Campos-Silva, Carmen
Cáceres-Martell, Yaiza
Sánchez-Herrero, Estela
Sandúa, Amaia
Beneítez-Martínez, Alexandra
González, Álvaro
Provencio, Mariano
Romero, Atocha
Jara-Acevedo, Ricardo
Yáñez-Mó, María
Valés-Gómez, Mar
author_role author
author2 Cáceres-Martell, Yaiza
Sánchez-Herrero, Estela
Sandúa, Amaia
Beneítez-Martínez, Alexandra
González, Álvaro
Provencio, Mariano
Romero, Atocha
Jara-Acevedo, Ricardo
Yáñez-Mó, María
Valés-Gómez, Mar
author2_role author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Ministerio de Ciencia, Innovación y Universidades (España)
Agencia Estatal de Investigación (España)
Comunidad de Madrid
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Extracellular vesicles
Cancer
Colloids
Flocculation
ELISA
Flow cytometry
Liquid biopsy
topic Extracellular vesicles
Cancer
Colloids
Flocculation
ELISA
Flow cytometry
Liquid biopsy
description [Background]: Extracellular vesicles (EVs), released by most cell types, provide an excellent source of biomarkers in biological fluids. However, in order to perform validation studies and screenings of patient samples, it is still necessary to develop general techniques permitting rapid handling of small amounts of biological samples from large numbers of donors.
publishDate 2022
dc.date.none.fl_str_mv 2022
2024
2024
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/346346
url http://hdl.handle.net/10261/346346
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/RTI2018-093569-B-I00
info:eu-repo/grantAgreement/AEI//RED2018-102411-T
S2017/BMD-3733-2/IMMUNOTHERCAN
IND2019/BMD-17258
https://doi.org/10.1186/s12951-022-01256-5
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Scientific Research Publishing
publisher.none.fl_str_mv Scientific Research Publishing
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15.812455

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