Jump to main content
Jump to site search

Issue 11, 2014
Previous Article Next Article

Microfluidic device (ExoChip) for on-chip isolation, quantification and characterization of circulating exosomes

Author affiliations

Abstract

Membrane bound vesicles, including microvesicles and exosomes, are secreted by both normal and cancerous cells into the extracellular space and in blood circulation. These circulating extracellular vesicles (cirEVs) and exosomes in particular are recognized as a potential source of disease biomarkers. However, to exploit the use of circulatory exosomes as a biomarker, a rapid, high-throughput and reproducible method is required for their isolation and molecular analysis. We have developed a simple, low cost microfluidic-based platform to isolate cirEVs enriched in exosomes directly from blood serum allowing simultaneous capture and quantification of exosomes in a single device. To capture specific exosomes, we employed “ExoChip”, a microfluidic device fabricated in polydimethylsiloxane (PDMS) and functionalized with antibodies against CD63, an antigen commonly overexpressed in exosomes. Subsequent staining with a fluorescent carbocyanine dye (DiO) that specifically labels the exosomes, we quantitated exosomes using a standard plate-reader. Ten independent ExoChip experiments performed using serum obtained from five pancreatic cancer patients and five healthy individuals revealed a statistically significant increase (2.34 ± 0.31 fold, p < 0.001) in exosomes captured in cancer patients when compared to healthy individuals. Exosomal origins of ExoChip immobilized vesicles were further confirmed using immuno-electron-microscopy and Western blotting. In addition, we demonstrate the ability of ExoChip to recover exosomes with intact RNA enabling profiling of exosomal-microRNAs through openarray analysis, which has potential applications in biomarker discovery. Based on our findings, ExoChip is a well suited platform to be used as an exosome-based diagnostic and research tool for molecular screening of human cancers.

Graphical abstract: Microfluidic device (ExoChip) for on-chip isolation, quantification and characterization of circulating exosomes

Back to tab navigation

Supplementary files

Publication details

The article was received on 30 Jan 2014, accepted on 19 Mar 2014 and first published on 20 Mar 2014


Article type: Paper
DOI: 10.1039/C4LC00136B
Author version available: Download Author version (PDF)
Citation: Lab Chip, 2014,14, 1891-1900
  •   Request permissions

    Microfluidic device (ExoChip) for on-chip isolation, quantification and characterization of circulating exosomes

    S. S. Kanwar, C. J. Dunlay, D. M. Simeone and S. Nagrath, Lab Chip, 2014, 14, 1891
    DOI: 10.1039/C4LC00136B

Search articles by author

Spotlight

Advertisements