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Dramatic enhancement of the detection limits of bioassays via ultrafast deposition of polydopamine

Nature Biomedical Engineering volume 1, Article number: 0082 (2017) Cite this article

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Abstract

The ability to detect biomarkers with ultrahigh sensitivity radically transformed biology and disease diagnosis. However, owing to incompatibilities with infrastructure in current biological and medical laboratories, recent innovations in analytical technology have not yet been adopted broadly. Here, we report a simple, universal ‘add-on’ technology (dubbed EASE) that converts the ordinary sensitivities of common bioassays to extraordinary ones, and that can be directly plugged into the routine practices of current research and clinical laboratories. The assay relies on the bioconjugation capabilities and ultrafast and localized deposition of polydopamine at the target site, which permit a large number of reporter molecules to be captured and lead to detection-sensitivity enhancements exceeding three orders of magnitude. The application of EASE in the ELISA-based detection of the HIV antigen in blood from patients leads to a sensitivity lower than 3 fg ml−1. We also show that EASE allows for the direct visualization, in tissues, of the Zika virus and of low-abundance biomarkers related to neurological diseases and cancer immunotherapy.

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Figure 1: HRP-accelerated dopamine polymerization and deposition.
Figure 2: IHC-EASE single-cell staining.
Figure 3: IF-EASE cell staining.
Figure 4: Ultrasensitive suspension microarray enabled by EASE.
Figure 5: ELISA and lateral flow strips with EASE.
Figure 6: Early diagnosis of HIV in patient blood samples using ELISA-EASE.
Figure 7: Sensitive imaging of ZIKV in placenta and PD-L1 in FFPE pancreatic tumour specimens.

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Acknowledgements

This work was supported in part by the NIH (R21CA192985, R01AI100989, AI083019, AI104002, and AI060389) and the Department of Bioengineering at the University of Washington. J.L. thanks the Howard Hughes Medical Institute for a student fellowship. We are also grateful to B. Lutz and D. Leon for help with the lateral flow test, and P. Zrazhevskiy for discussions on immunostaining.

Author information

Authors and Affiliations

  1. Department of Bioengineering, University of Washington, Seattle, 98195, Washington, USA

    Junwei Li, Wanyi Tai & Xiaohu Gao

  2. Department of Pharmacology, University of Washington, Seattle, 98105, Washington, USA

    Madison A. Baird & Larry S. Zweifel

  3. Department of Immunology, University of Washington, Seattle, 98105, Washington, USA

    Michael A. Davis & Michael Gale Jr

  4. Center for Innate Immunity and Immune Disease, University of Washington, Seattle, 98105, Washington, USA

    Michael A. Davis, Kristina M. Adams Waldorf, Michael Gale Jr & Lakshmi Rajagopal

  5. Department of Psychiatry and Behavioral Sciences, University of Washington, Seattle, 98105, Washington, USA

    Larry S. Zweifel

  6. Department of Obstetrics and Gynecology, University of Washington, Seattle, 98105, Washington, USA

    Kristina M. Adams Waldorf

  7. Department of Global Health, University of Washington, Seattle, 98105, Washington, USA

    Kristina M. Adams Waldorf, Michael Gale Jr & Lakshmi Rajagopal

  8. Sahlgrenska Academy, University of Gothenburg, Medicinaregatan 3, Gothenburg, 41390, Sweden

    Kristina M. Adams Waldorf

  9. Department of Pediatrics, University of Washington, Seattle, 98105, Washington, USA

    Lakshmi Rajagopal

  10. Center for Global Infectious Disease Research, Seattle Children’s Research Institute, Seattle, 98101, Washington, USA

    Lakshmi Rajagopal

  11. Fred Hutchinson Cancer Research Center, Program in Immunology, 1100 Fairview Avenue N, Seattle, 98109, Washington, USA

    Robert H. Pierce

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Contributions

J.L. and X.G. conceived the idea and designed the project. J.L. and W.T. performed the majority of the experiments, with help from M.A.B. and L.S.Z. for CRF imaging in the brain, from M.A.D., K.M.A.W., M.G. and L.R. for ZIKA imaging, and R.H.P. for PD-L1 imaging. All authors were involved in data analysis. J.L., L.S.Z., K.M.A.W., M.G., L.R., R.H.P. and X.G. wrote the paper.

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Correspondence to Xiaohu Gao.

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The authors declare no competing financial interests.

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Li, J., Baird, M., Davis, M. et al. Dramatic enhancement of the detection limits of bioassays via ultrafast deposition of polydopamine. Nat Biomed Eng 1, 0082 (2017). https://doi.org/10.1038/s41551-017-0082

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