Abstract
Tumor-derived extracellular vesicles (EVs) have become important biomarkers of liquid biopsies for precision medicine. However, the clinical application of EVs has been limited due to the lack of EV isolation practical technology applicable to clinical environments. Here, we report an innovative EV isolation method, which is quick and simple, and facilitates high-yield and high-purity EV isolation from blood. Introducing a cationic polymer in plasma resulted in rapid clustering of anionic EVs and a chaotropic agent can separate EVs from these clusters. Isolated EVs were characterized in terms of size distribution, morphology, surface protein markers, and exosomal RNA. Through performance comparison with various methods, including ultracentrifugation (UC), the present method delivered the highest recovery rate (~20 folds that of UC) and purity ratio (3.5 folds that of UC) of EVs in a short period of time (<20 min). The proposed method is expected to be used in basic and applied research on EV isolation and in clinical applications.
Highlights
Exosomes are nanosized vesicles (30–220 nm) extracellular vesicles (EVs), which, play a significant role in the delivery of signaling molecules for cell–cell communication [1,2,3,4] and in the mediation of pathological signaling [5,6,7]
PLL, is a protein composed of amino acids, guanidium thiocyanate (GuTc) unfolds and dissolves the amino acid chains of PLL to allow resuspension of EVs in liquid
Despite the clinical significance of EVs, technical advances in EV isolation and downstream RNA and protein analysis have been remained a major challenge in the field of clinical oncology
Summary
Exosomes are nanosized vesicles (30–220 nm) extracellular vesicles (EVs), which, play a significant role in the delivery of signaling molecules for cell–cell communication [1,2,3,4] and in the mediation of pathological signaling [5,6,7]. EVs including exosomes are shed from cells into body fluids such as blood and urine. Owing to the protein-lipid membrane of EVs, internal nucleic acids are securely preserved in body fluids [1,2]. Despite the potential significance of exosomes, they remain to be challenging analytes mainly due to the lack of EV isolation technology. The technical difficulty in EVs isolation arise from their unique characteristics such as nanoscale size, near neutral buoyancy, and containing excessive proteins and lipids in body fluids [10]
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