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首页> 外文期刊>Journal of Extracellular Vesicles >Cross‐flow microfiltration for isolation, selective capture and release of liposarcoma extracellular vesicles
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Cross‐flow microfiltration for isolation, selective capture and release of liposarcoma extracellular vesicles

机译:用于分离,选择性捕获和释放脂质瘤细胞外囊泡的交叉流量微滤

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We present a resource‐efficient approach to fabricate and operate a micro‐nanofluidic device that uses cross‐flow filtration to isolate and capture liposarcoma derived extracellular vesicles (EVs). The isolated extracellular vesicles were captured using EV‐specific protein markers to obtain vesicle enriched media, which was then eluted for further analysis. Therefore, the micro‐nanofluidic device integrates the unit operations of size‐based separation with CD63 antibody immunoaffinity‐based capture of extracellular vesicles in the same device to evaluate EV‐cargo content for liposarcoma. The eluted media collected showed ~76% extracellular vesicle recovery from the liposarcoma cell conditioned media and ~32% extracellular vesicle recovery from dedifferentiated liposarcoma patient serum when compared against state‐of‐art extracellular vesicle isolation and subsequent quantification by ultracentrifugation. The results reported here also show a five‐fold increase in amount of critical liposarcoma‐relevant extracellular vesicle cargo obtained in 30?min presenting a significant advance over existing state‐of‐art.
机译:我们提出了一种资源有效的方法来制造和操作微纳米流体装置,其使用横流过滤来分离和捕获脂质瘤衍生的细胞外囊泡(EVS)。使用EV特异性蛋白质标记物捕获分离的细胞外囊泡,得到浓缩培养基,然后洗脱进一步分析。因此,微纳米流体装置与基于CD63抗体免疫亲和基于CD63抗体免疫亲和的细胞外囊泡的单位操作集成了同一装置中的基于CD63抗体的分离,以评估脂质瘤的EV-FAR含量。收集的洗脱培养基显示脂肪瘤细胞条件培养基中的〜76%的细胞外囊泡回收,并且在与现有技术的细胞外囊泡分离和随后通过超速离心进行定量时,从消除的脂质糖尿病患者血清中恢复〜32%的细胞外囊泡回收。在此报告的结果还显示了在30?min中获得的临界脂肪脂肪瘤相关细胞内囊泡货物量的五倍增加,提出了对现有最先进的。

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