Microfluidic device for on-chip isolation and detection of circulating exosomes in blood of breast cancer patients

Abstract

Tumor-derived circulating exosomes have been recognized as a promising biomarker source for cancer diagnosis via a less invasive procedure. The integration of isolation and detection of exosomes in routine clinical settings is still challenging. In this study, we developed a new microfluidic device for immunomagnetic separation and detection of blood exosomes in situ. The microfluidic device may empower the integration of target exosome analysis via high surface to volume ratios of immunomagnetic beads and highly precise fluid control with the aid of microvalves. The obtained microfluidic device was capable of on-chip isolation and detection of circulating exosomes within 1.5 h. The captured exosomes could be directly visualized with an inverted fluorescence microscope in situ by tetramethylbenzidine-based colorimetric sensing. It was revealed that a statistically significant increase ( p < 0.01) in EpCAM-positive exosomes was captured for cancer patients (n = 10) on the device when compared to healthy individuals (n = 10). The device also demonstrated high predicting accuracy for tumor exosomal markers with a sensitivity of 90% and a specificity of >95% using receiver operating characteristic curves. The microfluidic device might provide a new platform to assist cancer diagnosis and molecular classification in an automated and simple fashion.

Rapid Capture of Cancer Extracellular Vesicles by Lipid Patch Microarrays

“…It was shown that the platform can reliably target EVs highly accurate and specific, out of a background of complex media and in mixtures containing off‐target types of EVs. The obtained sensitivity is matching that of other ultrasensitive methods (comparison to selected EV capture methods in Table S1, Supporting Information), [ 74–81 ] spanning the whole range of physiological relevant EV concentrations. Furthermore, the lipid arrays were also demonstrated to retain the RNA (and thus probably the other biomaterial cargo) of EVs in the system, which shows the platforms potential to isolate RNA for downstream analysis.…”

Towards nanovesicle-based disease diagnostics: a rapid single-step exosome assay within one hour throughin situimmunomagnetic extraction and nanophotonic label-free detection

“…Magnetic microbeads have been widely used for the separation of nucleic acids, protein, and EVs from clinic samples. [19][20][21][22][23][24][25][26][27] Although the microbeads allow fast and efficient collection of beads, the high material loss of microscale beads significantly quenches the resonance of the PC when the beads are immobilized. In contrast, the MNPs, whose diameter are much smaller than the operation wavelength, can be used to enhance the PC output without deteriorating the resonance mode.…”

“…16,17 Alternatively, immunomagnetic methods using functionalized magnetic micro-and nanoparticles can enable efficient, fast, and specific extractions of EVs. [18][19][20][21][22][23][24][25][26][27] Following an extraction process, the EV samples need to be eluted and detected using an EV sensing approach. For example, nanoparticle tracking analysis can be used to obtain EVs' size and phenotype.…”

Current Search through Liquid Biopsy of Effective Biomarkers for Early Cancer Diagnosis into the Rich Cargoes of Extracellular Vesicles

“…In both cases, the breast cancer cells-derived exosomes depicted a much higher MUC1 fluorescence, indicating the potential use of this method for further diagnosis of breast cancer. Chen et al [ 64 ] developed a new microfluidic device for immunomagnetic separation and detection of circulating exosomes in blood of breast cancer patients, able of on-chip isolation and detection of circulating exosomes within 1.5 h. A statistically significant increase ( p < 0.01) in EpCAM-positive exosomes was captured for breast cancer patients ( n = 10), when compared to healthy individuals ( n = 10). The device demonstrated high predicting accuracy for tumor exosomal markers with a sensitivity of 90% and a specificity of > 95%, providing a new automated platform to assist BC diagnosis.…”