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Characterization of Human Glioblastoma versus Normal Plasma-Derived Extracellular Vesicles Preisolated by Differential Centrifugation Using Cyclical Electrical Field-Flow Fractionation.
Analytical Chemistry ( IF 6.7 ) Pub Date : 2020-06-23 , DOI: 10.1021/acs.analchem.0c01373 Farhad Shiri 1 , Bruce K Gale 1 , Himanshu Sant 1 , Gina T Bardi 2 , Joshua L Hood 2, 3 , Kevin E Petersen 1
Analytical Chemistry ( IF 6.7 ) Pub Date : 2020-06-23 , DOI: 10.1021/acs.analchem.0c01373 Farhad Shiri 1 , Bruce K Gale 1 , Himanshu Sant 1 , Gina T Bardi 2 , Joshua L Hood 2, 3 , Kevin E Petersen 1
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Although many properties for small extracellular vesicles (sEVs, formerly termed “exosomes”) isolated at ∼100 000g are known, a wide range of values are reported for their electrophoretic mobility (EM) measurements. This paper reports for the first time the effect of dilution on the EM of U87 glioblastoma cell-derived and plasma-derived sEVs and medium size EVs (mEVs, commonly termed “oncosomes”) preisolated by differential centrifugation. Furthermore, the effect of resalting on the EM of sEVs and mEVs was evaluated. The EM of U87 sEVs and U87 mEVs showed an increase as the salt concentration decreased to 0.005% of the initial salt concentration. However, for the plasma sEVs and plasma mEVs, the electrophoretic mobility increased as the salt concentration decreased to 0.01% of the initial salt concentration and then increased to its initial value when the salt concentration decreased to 0.005% of the initial salt concentration. For both U87 and plasma sEVs and mEVs, the EM remained almost constant when the concentration of the particles changed and the salt concentration was kept the same as its initial value. This indicates that the EM of EVs is only a function of the salt concentration of the buffer and is independent of the concentration of the particles. The sEVs and mEVs were separated with cyclical ElFFF for the first time. The results indicate that ElFFF was able to fractionate the EVs, and a crescent-shaped trend was found for the retention time when the applied AC voltage was altered (increased).
中文翻译：
人胶质母细胞瘤与正常血浆来源的细胞外囊泡的表征，使用循环电场流分馏技术通过差异离心法进行预分离。
尽管分离出约100000 g的小细胞外囊泡（sEVs，以前称为“外来体”）的许多特性众所周知，据报道其电泳迁移率（EM）测量值范围广泛。本文首次报道了稀释对通过离心分离法分离的U87胶质母细胞瘤细胞和血浆衍生的sEV和中等大小的EV（mEV，通常称为“癌体”）EM的影响。此外，评估了转盐对sEV和mEV的EM的影响。当盐浓度降低到初始盐浓度的0.005％时，U87 sEV和U87 mEV的EM显示增加。然而，对于血浆sEV和血浆mEV，当盐浓度降低到初始盐浓度的0.01％时，电泳迁移率增加，而当盐浓度降低到初始盐浓度的0.005％时，电泳迁移率增加到其初始值。对于U87和血浆sEV和mEV，当颗粒浓度改变并且盐浓度保持与其初始值相同时，EM几乎保持恒定。这表明EV的EM仅是缓冲液的盐浓度的函数，并且与颗粒的浓度无关。sEV和mEV首次用周期性ElFFF分离。结果表明，ElFFF能够分馏电动汽车，并且当施加的交流电压发生变化（增加）时，保留时间呈新月形趋势。这表明EV的EM仅是缓冲液的盐浓度的函数，并且与颗粒的浓度无关。sEV和mEV首次用周期性ElFFF分离。结果表明，ElFFF能够分馏电动汽车，并且当施加的交流电压发生变化（增加）时，保留时间呈新月形趋势。这表明EV的EM仅是缓冲液的盐浓度的函数，并且与颗粒的浓度无关。sEV和mEV首次用周期性ElFFF分离。结果表明，ElFFF能够分馏电动汽车，并且当施加的交流电压发生变化（增加）时，保留时间呈新月形趋势。
更新日期：2020-07-21
中文翻译：
人胶质母细胞瘤与正常血浆来源的细胞外囊泡的表征，使用循环电场流分馏技术通过差异离心法进行预分离。
尽管分离出约100000 g的小细胞外囊泡（sEVs，以前称为“外来体”）的许多特性众所周知，据报道其电泳迁移率（EM）测量值范围广泛。本文首次报道了稀释对通过离心分离法分离的U87胶质母细胞瘤细胞和血浆衍生的sEV和中等大小的EV（mEV，通常称为“癌体”）EM的影响。此外，评估了转盐对sEV和mEV的EM的影响。当盐浓度降低到初始盐浓度的0.005％时，U87 sEV和U87 mEV的EM显示增加。然而，对于血浆sEV和血浆mEV，当盐浓度降低到初始盐浓度的0.01％时，电泳迁移率增加，而当盐浓度降低到初始盐浓度的0.005％时，电泳迁移率增加到其初始值。对于U87和血浆sEV和mEV，当颗粒浓度改变并且盐浓度保持与其初始值相同时，EM几乎保持恒定。这表明EV的EM仅是缓冲液的盐浓度的函数，并且与颗粒的浓度无关。sEV和mEV首次用周期性ElFFF分离。结果表明，ElFFF能够分馏电动汽车，并且当施加的交流电压发生变化（增加）时，保留时间呈新月形趋势。这表明EV的EM仅是缓冲液的盐浓度的函数，并且与颗粒的浓度无关。sEV和mEV首次用周期性ElFFF分离。结果表明，ElFFF能够分馏电动汽车，并且当施加的交流电压发生变化（增加）时，保留时间呈新月形趋势。这表明EV的EM仅是缓冲液的盐浓度的函数，并且与颗粒的浓度无关。sEV和mEV首次用周期性ElFFF分离。结果表明，ElFFF能够分馏电动汽车，并且当施加的交流电压发生变化（增加）时，保留时间呈新月形趋势。
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