Electrokinetic Evaluation of Individual Exosomes by On-Chip Microcapillary Electrophoresis with Laser Dark-Field Microscopy

Kato, Kenichi; Kobayashi, Masashi; Hanamura, Nami; Akagı, Takanori; Kosaka, Nobuyoshi; Ochiya, Takahiro; Ichikı, Takanori

doi:10.7567/jjap.52.06gk10

Cited by 40 publications

(30 citation statements)

References 35 publications

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“…The antibody binding on the cell surface was detected on a microcapillary electrophoresis (μCE) chip by measuring changes in the electrophoretic velocity of cells by particle tracking velocimetry. In analogy, the extended use of on-chip immunoelectrophoresis is expected to be a promising method for the surface marker analysis of individual EVs, but it requires some technical problems to be overcome such as the implementation of an appropriate imaging system that enables the tracking of the electrophoretic migration of individual vesicles smaller than the wavelength of light [ 28 , 29 ].…”

Section: Introductionmentioning

confidence: 99%

On-Chip Immunoelectrophoresis of Extracellular Vesicles Released from Human Breast Cancer Cells

et al. 2015

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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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Section: Introductionmentioning

confidence: 99%

On-Chip Immunoelectrophoresis of Extracellular Vesicles Released from Human Breast Cancer Cells

et al. 2015

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“…The lipid type in exosome bilayer, surface antigen type and density were found to be determining factors for the exome’s zeta potential. Using this knowledge, the authors managed investigated the correlation between the measured zeta potential of the exosomes and their cell of origin (Kato 2013) (Figure 4). …”

Section: - Detection Of Ev-containing Samplesmentioning

confidence: 99%

Microfluidic approaches for isolation, detection, and characterization of extracellular vesicles: Current status and future directions

Gholizadeh

Draz

Zarghooni

et al. 2017

Biosensors and Bioelectronics

178

113

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Extracellular vesicles (EVs) are cell-derived vesicles present in body fluids that play an essential role in various cellular processes, such as intercellular communication, inflammation, cellular homeostasis, survival, transport, and regeneration. Their isolation and analysis from body fluids have a great clinical potential to provide information on a variety of disease states such as cancer, cardiovascular complication and inflammatory disorders. Despite increasing scientific and clinical interest in this field, at the time of writing there are still no standardized procedures available for the purification, detection, and characterization of EVs. Advances in microfluidics allow for chemical sampling with increasingly high spatial resolution and under precise manipulation down to single molecule level. In this review, our objective is to give a brief overview on the working principle and examples of the isolation and detection methods with the potential to be used for extracellular vesicles. This review will also highlight the integrated on-chip systems for isolation and characterization of EVs.

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“…Zeta potential of individual exosomes was evaluated by a procedure described previously [17][18][19]. Briefly, after of a sample chip, the chip was placed on the stage of an inverted microscope, and then electrodes were dipped into the reservoirs at both ends of the microchannel.…”

Section: Zeta Potential Measurementmentioning

confidence: 99%

“…Recently, we have developed an on-chip microcapillary electrophoresis ( CE) platform equipped with a laser-dark field microscope that enables the zeta potential of individual nanobioparticles to be obtained [17][18][19]. Additionally, an analytical method for profiling the surface proteins of individual exosomes that employs the principle of on-chip particle immunoelectrophoresis has been proposed [19].…”

Section: Introductionmentioning

confidence: 99%

Measurement of Individual Nanobioparticles on Microfluidic Chips by Laser Dark-field Imaging

Akagı

Hanamura

Ichikı

2015

J. Photopol. Sci. Technol.

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A microfluidic-chip-based analysis platform specially tuned for characterizing individual nanobio-particles has been developed. Using this platform, both artificial nanobioparticles (polyion complex vesicles) and cell-secreted exosomes with diameter down to 50 nm can be observed. The zeta potential distribution of the exosomes was obtained by measuring the zeta potential of individual exosomes. Since the number of surface molecules on nanobioparticles can be estimated by applying a particle immunoelectrophoresis method, this platform is promising for obtaining profiles of heterogeneous nanobioparticles including nanoDDS carriers and exosomes.

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Electrokinetic Evaluation of Individual Exosomes by On-Chip Microcapillary Electrophoresis with Laser Dark-Field Microscopy

Cited by 40 publications

References 35 publications

On-Chip Immunoelectrophoresis of Extracellular Vesicles Released from Human Breast Cancer Cells

On-Chip Immunoelectrophoresis of Extracellular Vesicles Released from Human Breast Cancer Cells

Microfluidic approaches for isolation, detection, and characterization of extracellular vesicles: Current status and future directions

Measurement of Individual Nanobioparticles on Microfluidic Chips by Laser Dark-field Imaging

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