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Label-free detection and molecular profiling of exosomes with a nano-plasmonic sensor

Nature Biotechnology volume 32, pages 490–495 (2014)Cite this article

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Abstract

Exosomes show potential for cancer diagnostics because they transport molecular contents of the cells from which they originate. Detection and molecular profiling of exosomes is technically challenging and often requires extensive sample purification and labeling. Here we describe a label-free, high-throughput approach for quantitative analysis of exosomes. Our nano-plasmonic exosome (nPLEX) assay is based on transmission surface plasmon resonance through periodic nanohole arrays. Each array is functionalized with antibodies to enable profiling of exosome surface proteins and proteins present in exosome lysates. We show that this approach offers improved sensitivity over previous methods, enables portable operation when integrated with miniaturized optics and allows retrieval of exosomes for further study. Using nPLEX to analyze ascites samples from ovarian cancer patients, we find that exosomes derived from ovarian cancer cells can be identified by their expression of CD24 and EpCAM, suggesting the potential of exosomes for diagnostics.

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Figure 1: Label-free detection of exosomes with nPLEX sensor.
Figure 2: Exosome quantification and protein profiling with nPLEX.
Figure 3: Molecular profiling of ovarian cancer protein markers.
Figure 4: Profiling of ovarian cancer patient exosomes with nPLEX.

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Acknowledgements

The authors thank S. Skates (Massachusetts General Hospital) for helpful discussion on statistical analyses; M. Birrer for facilitating sample collection; K. Joyes for reviewing the manuscript. This work was supported in part by US National Institutes of Health (NIH) grants R01-HL113156 (H.L.), R01-EB010011 (R.W.), R01-EB00462605A1 (R.W.), T32CA79443 (R.W.), K12CA087723-11A1 (C.M.C) and National Heart, Lung, and Blood Institute contract HHSN268201000044C (R.W.). The device was fabricated using the facilities at the Center for Nanoscale Systems (CNS) at Harvard University (National Science Foundation award ECS-0335765).

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  1. Hyungsoon Im and Huilin Shao: These authors contributed equally to this work.

Authors and Affiliations

  1. Center for Systems Biology, Massachusetts General Hospital, Boston, Massachusetts, USA

    Hyungsoon Im, Huilin Shao, Yong Il Park, Vanessa M Peterson, Cesar M Castro, Ralph Weissleder & Hakho Lee

  2. Department of Systems Biology, Harvard Medical School, Boston, Massachusetts, USA

    Ralph Weissleder

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  1. Hyungsoon Im
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Contributions

H.I., H.S., R.W. and H.L. designed the research. C.M.C. and R.W. designed the clinical study. H.I., H.S., Y.I.P., V.M.P. and C.M.C. performed the research. V.M.P. and C.M.C. collected the clinical samples. H.I., H.S., R.W. and H.L. analyzed data. H.I., H.S., C.M.C., R.W. and H.L. wrote the paper.

Corresponding authors

Correspondence to Ralph Weissleder or Hakho Lee.

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The authors declare no competing financial interests.

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Supplementary Figures 1–15 and Supplementary Tables 1–4 (PDF 2898 kb)

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Im, H., Shao, H., Park, Y. et al. Label-free detection and molecular profiling of exosomes with a nano-plasmonic sensor. Nat Biotechnol 32, 490–495 (2014). https://doi.org/10.1038/nbt.2886

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