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Size-Exclusion Chromatography

2024-08Artigo científico Acesso aberto
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dc.contributor.authorGrenhas, Madalena
dc.contributor.authorLopes, Raquel
dc.contributor.authorFerreira, Bruna Velosa
dc.contributor.authorBarahona, Filipa
dc.contributor.authorJoão, Cristina
dc.contributor.authorCarneiro, Emilie Arnault
dc.contributor.institutionNOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM)
dc.contributor.pblMDPI - Multidisciplinary Digital Publishing Institute
dc.date.accessioned2024-09-04T22:20:22Z
dc.date.available2024-09-04T22:20:22Z
dc.date.issued2024-08
dc.descriptionFunding Information: The authors express their gratitude to the Electron Microscopy Unit of Instituto Gulbenkian de Ci\u00EAncia (IGC) for the acquisition of the EV images. Additionally, we extend our appreciation to Carolina Pestana MSc for her contribution to the statistical analysis of this study, to the Systems Oncology Research Group (Champalimaud Foundation) for granting access to the required equipment for this study, and to the Champalimaud Foundation for funding. Publisher Copyright: © 2024 by the authors.
dc.description.abstractSmall extracellular vesicles (EVs) play a pivotal role in intercellular communication across various physiological and pathological contexts. Despite their growing significance as disease biomarkers and therapeutic targets in biomedical research, the lack of reliable isolation techniques remains challenging. This study characterizes vesicles that were isolated from conditioned culture media (CCM) sourced from three myeloma cell lines (MM.1S, ANBL-6, and ALMC-1), and from the plasma of healthy donors and multiple myeloma patients. We compared the efficacy, reproducibility, and specificity of isolating small EVs using sucrose cushion ultracentrifugation (sUC) vs. ultrafiltration combined with size-exclusion chromatography (UF-SEC). Our results demonstrate that UF-SEC emerges as a more practical, efficient, and consistent method for EV isolation, outperforming sUC in the yield of EV recovery and exhibiting lower variability. Additionally, the comparison of EV characteristics among the three myeloma cell lines revealed distinct biomarker profiles. Finally, our results suggest that HBS associated with Tween 20 improves EV recovery and preservation over PBS. Standardization of small EV isolation methods is imperative, and our comparative evaluation represents a significant step toward achieving this goal.en
dc.description.versionpublishersversion
dc.description.versionpublished
dc.format.extent5047414
dc.identifier.doi10.3390/ijms25158496
dc.identifier.issn1661-6596
dc.identifier.otherPURE: 98805870
dc.identifier.otherPURE UUID: ac724166-e3f7-4610-b67a-cfad56bd730c
dc.identifier.otherScopus: 85200835150
dc.identifier.otherPubMed: 39126063
dc.identifier.urihttp://hdl.handle.net/10362/171170
dc.identifier.urlhttps://www.scopus.com/pages/publications/85200835150
dc.language.isoeng
dc.peerreviewedyes
dc.subjectextracellular vesicle isolation methods
dc.subjectextracellular vesicles
dc.subjectmultiple myeloma
dc.subjectmyeloma cell lines
dc.subjectnewly diagnosed multiple myeloma
dc.subjectplasma
dc.subjectsize-exclusion chromatography
dc.subjectsucrose cushion ultracentrifugation
dc.subjectultrafiltration
dc.subjectCatalysis
dc.subjectMolecular Biology
dc.subjectSpectroscopy
dc.subjectComputer Science Applications
dc.subjectPhysical and Theoretical Chemistry
dc.subjectOrganic Chemistry
dc.subjectInorganic Chemistry
dc.titleSize-Exclusion Chromatographyen
dc.title.subtitleA Path to Higher Yield and Reproducibility Compared to Sucrose Cushion Ultracentrifugation for Extracellular Vesicle Isolation in Multiple Myelomaen
dc.typejournal article
degois.publication.issue15
degois.publication.titleInternational Journal of Molecular Sciences
degois.publication.volume25
dspace.entity.typePublication
rcaap.rightsopenAccess

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