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dc.contributor.authorLeong, Sheng Yuanen_US
dc.contributor.authorOng, Hong Boonen_US
dc.contributor.authorTay, Hui Minen_US
dc.contributor.authorKong, Fangen_US
dc.contributor.authorUpadya, Meghaen_US
dc.contributor.authorGong, Lingyanen_US
dc.contributor.authorDao, Mingen_US
dc.contributor.authorDalan, Rinkooen_US
dc.contributor.authorHou, Han Weien_US
dc.identifier.citationLeong, S. Y., Ong, H. B., Tay, H. M., Kong, F., Upadya, M., Gong, L., Dao, M., Dalan, R. & Hou, H. W. (2022). Microfluidic size exclusion chromatography (μSEC) for extracellular vesicles and plasma protein separation. Small, 18(6), 2104470-.
dc.description.abstractExtracellular vesicles (EVs) are recognized as next generation diagnostic bio-markers due to their disease-specific biomolecular cargoes and importance in cell–cell communications. A major bottleneck in EV sample preparation is the inefficient and laborious isolation of nanoscale EVs (≈50–200 nm) from endogenous proteins in biological samples. Herein, a unique microfluidic platform is reported for EV-protein fractionation based on the principle of size exclusion chromatography (SEC). Using a novel rapid (≈20 min) replica molding technique, a fritless microfluidic SEC device (μSEC) is fabricated using thiol-ene polymer (UV glue NOA81, Young’s modulus ≈1 GPa) for high pressure (up to 6 bar) sample processing. Controlled on-chip nano-liter sample plug injection (600 nL) using a modified T-junction injector is first demonstrated with rapid flow switching response time (<1.5 s). Device performance is validated using fluorescent nanoparticles (50 nm), albumin, and breast cancer cells (MCF-7)-derived EVs. As a proof-of-concept for clinical applications, EVs are directly isolated from undiluted human platelet-poor plasma using μSEC and show distinct elution profiles between EVs and proteins based on nanoparticle particle analysis (NTA), Western blot and flow cytometry analysis. Overall, the optically transparent μSEC can be readily automated and integrated with EV detection assays for EVs manufacturing and clinical diagnostics.en_US
dc.description.sponsorshipNanyang Technological Universityen_US
dc.description.sponsorshipSingapore-MIT Alliance for Research and Technology (SMART)en_US
dc.relationING-000539 BIO IGNen_US
dc.relationING-001058 BIO IGNen_US
dc.rights© 2022 Wiley-VCH GmbH. All rights reserved.en_US
dc.subjectScience::Biological sciencesen_US
dc.titleMicrofluidic size exclusion chromatography (μSEC) for extracellular vesicles and plasma protein separationen_US
dc.typeJournal Articleen
dc.contributor.schoolSchool of Biological Sciencesen_US
dc.contributor.schoolSchool of Mechanical and Aerospace Engineeringen_US
dc.contributor.schoolLee Kong Chian School of Medicine (LKCMedicine)en_US
dc.contributor.organizationTan Tock Seng Hospitalen_US
dc.subject.keywordsExtracellular Vesiclesen_US
dc.subject.keywordsSize Exclusion Chromatographyen_US
dc.description.acknowledgementH.W.H. wrote the manuscript. All authors reviewed the manuscript. H.W.H. would like to acknowledge the kind financial support from SMART Innovation Centre (ING-000539 BIO IGN and ING-001058 BIO IGN), and NTU Startup grant. S.Y.L. and L.G. would like to acknowledge support from the NTU Research Scholarship.en_US
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