Abstract

Extracellular vesicles (EVs) are considered essential biomarkers in liquid biopsies. Despite intensive efforts aimed at employing EVs in a clinical setting, workable approaches are currently limited owing to the fact that EV-isolation technologies are still in a nascent stage. This study introduces a magnetic bead-based ion exchange platform for isolating EVs called ExoCAS-2 (exosome clustering and scattering). Owing to their negative charge, exosomes can easily adhere to magnetic beads coated with a polycationic polymer. Owing to the features of magnetic beads, exosomes can be easily processed via washing and elution steps and isolated with high purity and yield within 40 min. The present results confirmed the isolation of exosomes through analyses of size distribution, morphology, surface and internal protein markers, and exosomal RNA. Compared with the commercially available methods, the proposed method showed superior performance in terms of key aspects, including operation time, purity, and recovery rate. This highlights the potential of this magnetic bead-based ion exchange platform for isolating exosomes present in blood plasma.

Highlights

  • Nano-sized (30–160 nm) exosomes, discovered in 1983 [1,2], are a subset of extracellular vesicles (EVs)

  • Several exosomes were bound to the surfaces of PLL-coated magnetic beads

  • The PLL-coated magnetic beads were visualized by staining with FITC, whereas exosomes were stained with anti-CD 63 (Alexa Fluor 647)

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Summary

Introduction

Nano-sized (30–160 nm) exosomes, discovered in 1983 [1,2], are a subset of extracellular vesicles (EVs). They are shed from almost all cell types in the human body into body fluids, such as blood, urine, saliva, milk, semen, etc. Once considered to be cellular garbage packs, exosomes are recognized as important mediators of intercellular communication, transporting diverse cargo to other cells. Exosomes are considered to be closely associated with the pathological mechanisms underlying various diseases, including cancer, neurodegenerative disorders, multiple sclerosis, and inflammatory diseases, by virtue of their role in intercellular communication [9,10,11,12,13,14]

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