Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/154936
Title: Microfluidic size exclusion chromatography (μSEC) for extracellular vesicles and plasma protein separation
Authors: Leong, Sheng Yuan
Ong, Hong Boon
Tay, Hui Min
Kong, Fang
Upadya, Megha
Gong, Lingyan
Dao, Ming
Dalan, Rinkoo
Hou, Han Wei
Keywords: Science::Biological sciences
Science::Medicine
Issue Date: 2022
Source: Leong, 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-. https://dx.doi.org/10.1002/smll.202104470
Project: ING-000539 BIO IGN 
ING-001058 BIO IGN 
Journal: Small 
Abstract: Extracellular 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.
URI: https://hdl.handle.net/10356/154936
ISSN: 1613-6810
DOI: 10.1002/smll.202104470
Schools: School of Biological Sciences 
School of Mechanical and Aerospace Engineering 
Lee Kong Chian School of Medicine (LKCMedicine) 
Organisations: Tan Tock Seng Hospital
Rights: © 2022 Wiley-VCH GmbH. All rights reserved. This is the peer reviewed version of the following article: Leong, 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-. https://dx.doi.org/10.1002/smll.202104470, which has been published in final form at https://dx.doi.org/10.1002/smll.202104470. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:LKCMedicine Journal Articles
MAE Journal Articles
SBS Journal Articles

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