Open AccessA simple, high-throughput method of protein and label removal from extracellular vesicle samples
Author(s) -
Joshua A. Welsh,
Bryce Killingsworth,
Julia Kepley,
Tim Traynor,
Kathy McKin,
Jason E. Savage,
Deven Appel,
Kenneth Aldape,
Kevin Camphausen,
Jay A. Berzofsky,
Alexander R. Ivanov,
Ionita Ghiran,
Jennifer C. Jones
Publication year - 2021
Publication title -
nanoscale
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.038
H-Index - 224
eISSN - 2040-3372
pISSN - 2040-3364
DOI - 10.1039/d0nr07830a
Subject(s) - extracellular vesicles , centrifugation , filtration (mathematics) , reagent , chromatography , chemistry , throughput , extracellular vesicle , vesicle , fluorescence , nanotechnology , microvesicles , materials science , computer science , biochemistry , biology , microbiology and biotechnology , membrane , telecommunications , microrna , statistics , mathematics , gene , wireless , physics , quantum mechanics
Evidence continues to increase of the clinical utility extracellular vesicles (EVs) as translational biomarkers. While a wide variety of EV isolation and purification methods have been implemented, few techniques are high-throughput and scalable for removing excess fluorescent reagents (e.g. dyes, antibodies). EVs are too small to be recovered from routine cell-processing procedures, such as filtration or centrifugation. The lack of suitable methods for removing unbound labels, especially in optical assays, is a major roadblock to accurate EV phenotyping and utilization of EV assays in a translational or clinical setting. Therefore, we developed a method for using a multi-modal resin, referred to as EV-Clean, to remove unbound labels from EV samples, and we demonstrate improvement in flow cytometric EV analysis with the use of this EV-Clean method.