期刊
NANO LETTERS
卷 18, 期 7, 页码 4226-4232出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.nanolett.8b01184
关键词
Droplet microfluidics; exosomes; droplet digital ExoELISA; cancer diagnostics
类别
资金
- Innovation and Technology Fund [ITS/224/16]
- National Natural Science Foundation of China [81702100]
- Science and Technology Program of Guangzhou [201510010097]
- Major Program of Health Care and Innovation of Guangzhou [201704020213, 201604020015]
Exosomes shed by tumor cells have been recognized as promising biomarkers for cancer diagnostics due to their unique composition and functions. Quantification of low concentrations of specific exosomes present in very small volumes of clinical samples may be used for noninvasive cancer diagnosis and prognosis. We developed an immunosorbent assay for digital qualification of target exosomes using droplet microfluidics. The exosomes were immobilized on magnetic microbeads through sandwich ELISA complexes tagged with an enzymatic reporter that produces a fluorescent signal. The constructed beads were further isolated and encapsulated into a sufficient number of droplets to ensure only a single bead was encapsulated in a droplet. Our droplet-based single-exosome-counting enzyme-linked immunoassay (droplet digital ExoELISA) approach enables absolute counting of cancer-specific exosomes to achieve unprecedented accuracy. We were able to achieve a limit of detection (LOD) down to 10 enzyme-labeled exosome complexes per microliter (similar to 10(-17) M). We demonstrated the application of the droplet digital ExoELISA platform in quantitative detection of exosomes in plasma samples directly from breast cancer patients. We believe our approach may have the potential for early diagnosis of cancer and accelerate the discovery of cancer exosomal biomarkers for clinical diagnosis.
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