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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.

As recent advances in medicine rapidly unravel the genomic and proteomic signatures of disease development, progression and response to therapy, sensitive and quantitative analysis of disease biomarkers (such as DNA, RNA and proteins) has become increasingly important in the era of precision medicine, in which diagnostic and therapeutic decisions are tailored towards individual patients. In parallel, to address the challenge in sensitive and multiplexed biomarker analysis, a large variety of intricately designed imaging and detection technologies have also been developed in the past decade1,2,3,4,5,6,7,8,9,10,11. These enabling technologies—often leveraging on the unique properties of colloidal nanostructures (such as quantum dots, magnetic nanoparticles and plasmonic nanoparticles) and precisely engineered sensor devices (for example, nanowire sensors, cantilevers and microfluidic channels)—are so sensitive that their detection limits are commonly seen at the single-molecule level, and low-abundance targets such as circulating oligonucleotides, proteins, viruses and cells can be enumerated with polymerase chain reaction (PCR)-like sensitivity12,13,14,15,16,17,18. Despite these remarkable achievements in biotechnology laboratories, broad adoption of these technological innovations by biology and clinical laboratories has been limited, and consequently so has their impact19. This is a result of multiple factors, including complex protocols, the need for specialized reagents and equipment, and most importantly the requirement of different infrastructures, a disconnect that not only elevates the upfront cost but also reduces persistent output and cross-laboratory cross-platform consistency.

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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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  1. Junwei Li
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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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