Welsh Joshua A, Killingsworth Bryce, Kepley Julia, Traynor Tim, McKinnon Kathy, Savage Jason, Appel Deven, Aldape Kenneth, Camphausen Kevin, Berzofsky Jay A, Ivanov Alexander R, Ghiran Ionita H, Jones Jennifer C
Translational Nanobiology Section, Laboratory of Pathology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
Vaccine Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
Nanoscale. 2021 Feb 18;13(6):3737-3745. doi: 10.1039/d0nr07830a.
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.
细胞外囊泡（EVs）作为转化生物标志物的临床应用证据不断增加。虽然已经实施了各种各样的EV分离和纯化方法，但很少有技术能够高通量且可扩展地去除过量的荧光试剂（如染料、抗体）。EVs太小，无法从常规细胞处理程序（如过滤或离心）中回收。缺乏合适的方法来去除未结合的标记物，尤其是在光学检测中，这是在转化或临床环境中准确进行EV表型分析和利用EV检测的主要障碍。因此，我们开发了一种使用称为EV-Clean的多模态树脂从EV样品中去除未结合标记物的方法，并证明使用这种EV-Clean方法可改善流式细胞术对EV的分析。
