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利用聚酯、毛细管通道聚合物纤维相通过疏水相互作用层析法分离和纯化外泌体。

Exosome isolation and purification via hydrophobic interaction chromatography using a polyester, capillary-channeled polymer fiber phase.

机构信息

Department of Bioengineering, Life Sciences Facility, Clemson University, Clemson, SC, USA.

Department of Chemistry, Biosystems Research Complex, Clemson University, Clemson, SC, USA.

出版信息

Electrophoresis. 2019 Feb;40(4):571-581. doi: 10.1002/elps.201800417. Epub 2018 Dec 27.

Abstract

Extracellular vesicles, including microvesicles and exosomes, are lipidic membrane-derived vesicles that are secreted by most cell types. Exosomes, one class of these vesicles that are 30-100 nm in diameter, hold a great deal of promise in disease diagnostics, as they display the same protein biomarkers as their originating cell. For exosomes to become useful in disease diagnostics, and as burgeoning drug delivery platforms, they must be isolated efficiently and effectively without compromising their structure. Most current exosome isolation methods have practical problems including being too time-consuming and labor intensive, destructive to the exosomes, or too costly for use in clinical settings. To this end, this study examines the use of poly(ethylene terephthalate) (PET) capillary-channeled polymer (C-CP) fibers in a hydrophobic interaction chromatography (HIC) protocol to isolate exosomes from diverse matrices of practical concern. Initial results demonstrate the ability to isolate extracellular vesicles enriched in exosomes with comparable yields and size distributions on a much faster time scale when compared to traditional isolation methods. As a demonstration of the potential analytical utility of the approach, extracellular vesicle recoveries from cell culture milieu and a mock urine matrix are presented. The potential for scalable separations covering submilliliter spin-down columns to the preparative scale is anticipated.

摘要

细胞外囊泡,包括微囊泡和外泌体,是由大多数细胞类型分泌的脂质膜衍生的囊泡。外泌体是这些囊泡的一种,直径为 30-100nm,在疾病诊断中具有很大的应用潜力,因为它们显示出与起源细胞相同的蛋白质生物标志物。为了使外泌体在疾病诊断中变得有用,并作为新兴的药物输送平台,它们必须在不破坏其结构的情况下有效地进行分离。目前大多数外泌体分离方法都存在实际问题,包括耗时太长、劳动强度大、对囊泡具有破坏性,或者在临床环境中使用成本过高。为此,本研究探讨了在疏水性相互作用色谱(HIC)方案中使用聚对苯二甲酸乙二醇酯(PET)毛细管通道聚合物(C-CP)纤维从实际关注的各种基质中分离外泌体的用途。初步结果表明,与传统分离方法相比,该方法能够更快地分离出富含外泌体的细胞外囊泡,并且具有相当的产量和粒径分布。作为该方法潜在分析应用的演示,展示了从细胞培养环境和模拟尿液基质中回收细胞外囊泡的情况。预计该方法具有从亚毫升离心柱到制备规模的可扩展分离潜力。

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