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Exosome aggregation mediated stop‐flow paper‐based portable device for rapid exosome quantification

机译:外出聚集介导的止动纸基便携式装置,用于快速外出量化

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Abstract Exosome quantification is important for estimation of informative messengers (e.g., proteins, lipids, RNA, etc.) involving physiological and pathological effects. This work aimed to develop a simple and rapid distance‐based paper portable device using exosome‐capture vesicles (polydiacetylene conjugated with antiCD81) for exosome quantification in cell cultures. This novel concept relied on distinct aggregation of exosomes and exosome‐capture vesicles leading to different solvent migration. Distances of the migration were used as signal readouts, which could be detected by naked eye. PDA‐antiCD81 as exosome‐capture vesicles were optimized (e.g., size, reaction ratio, and concentration) and the paper designs were investigated (e.g., diameter of sample reservoir and lamination layer) to enhance the solvent stop‐flow effects. Finally, exosome screening on three cell culture samples (COLO1, MDA‐MB‐231, and HuR‐KO1 subclone) was demonstrated. The method could linearly measure exosome concentrations in correlation with solvent migration distances in the range of 10 6 –10 10 particles/mL ( R 2 ??0.98) from the cell culture samples. The exosome concentration measurements by the developed device were independently assessed by nanoparticle tracking analysis. Results demonstrated no statistically significant difference ( p ? 0.05) by t ‐test. This low‐cost and rapid device allows a portable platform for exosome quantification without the requirement of expensive equipment and expertise of operation. The developed device could potentially be useful for quantification of other biomarker‐related extracellular vesicles.
机译:抽象外来体定量为翔实信使(例如,蛋白质,脂质,RNA等)有关生理和病理作用的估计重要。这项工作的目的是开发用于在细胞培养物的外来体定量的简单和使用外来体捕获囊泡快速的基于距离的纸便携式设备(聚二乙炔与antiCD81缀合)。这种新颖的概念依赖于外来体和外来体捕获囊泡从而导致不同的溶剂迁移的不同聚集。迁移的距离被用作信号读出,其可以通过肉眼进行检测。 PDA-antiCD81作为外来体捕获囊泡进行优化(例如,大小,反应率,和浓度)以及纸张的设计进行了研究(例如,样本池和层压层的直径),以增强溶剂停流的效果。最后,在三种细胞培养物样品(COLO1,MDA-MB-231,和的HuR-KO1亚克隆)的外来体的筛选证实。该方法可线性地测量在相关外来体的浓度与溶剂迁移距离在10 6 -10 10个颗粒/ mL(R 2 - ; 0.98?)的范围从细胞培养物的样品。由开发的设备的外来体浓度测量是独立地由纳米粒子追踪分析评估。结果通过t检验表明无统计学显著差异(p&GT ;? 0.05)。这种低成本,快速的设备允许外来体定量便携式平台,而无需昂贵的设备和操作的专业知识的要求。该开发的设备可能是其他生物标志物相关的细胞外囊泡的定量是有用的。

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