On-chip immunoelectrophoresis of extracellular vesicles released from human breast cancer cells.
On-chip immunoelectrophoresis of extracellular vesicles released from human breast cancer cells.
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DOI：
10.1371/journal.pone.0123603
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Ichiki T
中科院分区：
文献类型：
--
作者：
Akagi T;Kato K;Kobayashi M;Kosaka N;Ochiya T;Ichiki T
细胞外囊泡（extracellular vesicles，EV）包括外泌体和微囊泡，在细胞生物学和医学领域引起了相当大的关注。为了更好地了解电动汽车和进一步探索其应用，生物纳米囊泡的分析方法的发展一直是必需的。特别地，考虑到EV的异质性，需要能够测量单个囊泡的方法。在这里，我们报告，芯片上的免疫电泳可以提供一个有用的方法，个别EV的差异蛋白表达谱。使用包括微毛细管电泳芯片和激光暗场显微成像系统的测量平台，对从MDA-MB-231人乳腺癌细胞的培养上清液收集的EV进行电泳实验。与正常免疫球蛋白G（IgG）同种型对照相比，与抗人CD 63（外来体和微囊泡标记物）抗体反应的EV的ζ电位分布显示出显著的正向偏移。因此，芯片免疫电泳可以灵敏地检测EV上的CD 63糖蛋白的过表达。此外，为了探索芯片上免疫电泳对癌症诊断的适用性，通过该方法分析了从小鼠肿瘤模型的血液中收集的EV。通过比较EV与正常IgG、抗人CD 63和抗人CD 44（癌症干细胞标记物）抗体免疫化学反应后的Zeta电位分布，在真实的样品中差异检测血液中循环的肿瘤来源的EV。结果表明，本方法是潜在的适用于液体活检，一个有前途的方法，以低侵入性的癌症诊断。
Extracellular vesicles (EVs) including exosomes and microvesicles have attracted considerable attention in the fields of cell biology and medicine. For a better understanding of EVs and further exploration of their applications, the development of analytical methods for biological nanovesicles has been required. In particular, considering the heterogeneity of EVs, methods capable of measuring individual vesicles are desired. Here, we report that on-chip immunoelectrophoresis can provide a useful method for the differential protein expression profiling of individual EVs. Electrophoresis experiments were performed on EVs collected from the culture supernatant of MDA-MB-231 human breast cancer cells using a measurement platform comprising a microcapillary electrophoresis chip and a laser dark-field microimaging system. The zeta potential distribution of EVs that reacted with an anti-human CD63 (exosome and microvesicle marker) antibody showed a marked positive shift as compared with that for the normal immunoglobulin G (IgG) isotype control. Thus, on-chip immunoelectrophoresis could sensitively detect the over-expression of CD63 glycoproteins on EVs. Moreover, to explore the applicability of on-chip immunoelectrophoresis to cancer diagnosis, EVs collected from the blood of a mouse tumor model were analyzed by this method. By comparing the zeta potential distributions of EVs after their immunochemical reaction with normal IgG, and the anti-human CD63 and anti-human CD44 (cancer stem cell marker) antibodies, EVs of tumor origin circulating in blood were differentially detected in the real sample. The result indicates that the present method is potentially applicable to liquid biopsy, a promising approach to the low-invasive diagnosis of cancer.
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影响因子：
46.9
作者：
Alvarez-Erviti, Lydia;Seow, Yiqi;Wood, Matthew J. A.
通讯作者：
Wood, Matthew J. A.
影响因子：
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作者：
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影响因子：
4
作者：
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通讯作者：
Kuo WP
影响因子：
8.8
作者：
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影响因子：
7.4
作者：
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通讯作者：
Lyerly HK