A simple, high-throughput method of protein and label removal from extracellular vesicle samples

一种从细胞外囊泡样品中去除蛋白质和标记物的简便、高通量方法

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作者:Joshua A Welsh ,Bryce Killingsworth ,Julia Kepley ,Tim Traynor ,Kathy McKinnon ,Jason Savage ,Deven Appel ,Kenneth Aldape ,Kevin Camphausen ,Jay A Berzofsky ,Alexander R Ivanov ,Ionita H Ghiran ,Jennifer C Jones

Abstract

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.

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