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An ultrasensitive universal detector based on neutralizer displacement

Nature Chemistry volume 4, pages 642–648 (2012)Cite this article

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Abstract

Diagnostic technologies that can provide the simultaneous detection of nucleic acids for gene expression, proteins for host response and small molecules for profiling the human metabolome will have a significant advantage in providing comprehensive patient monitoring. Molecular sensors that report changes in the electrostatics of a sensor's surface on analyte binding have shown unprecedented sensitivity in the detection of charged biomolecules, but do not lend themselves to the detection of small molecules, which do not carry significant charge. Here, we introduce the neutralizer displacement assay that allows charge-based sensing to be applied to any class of molecule irrespective of the analyte charge. The neutralizer displacement assay starts with an aptamer probe bound to a neutralizer. When analyte binding occurs the neutralizer is displaced, which results in a dramatic change in the surface charge for all types of analytes. We have tested the sensitivity, speed and specificity of this system in the detection of a panel of molecules: (deoxy)ribonucleic acid, ribonucleic acid, cocaine, adenosine triphosphate and thrombin.

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Figure 1: Summary of the NDA and sensor chips utilized for testing.
Figure 2: NDA small molecule detection: ATP and cocaine.
Figure 3: NDA nucleic acid and bacterial detection.
Figure 4: NDA protein detection.

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Acknowledgements

We acknowledge the Natural Sciences and Engineering Research Council (Discovery Grant to S.O.K.), Mitacs (Elevate Fellowship to J.D.) and the Defense Advanced Research Projects Agency (DXoD programme funding to S.O.K. and E.H.S.) for their support of this work.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, Canada

    Jagotamoy Das, Kristin B. Cederquist, Alexandre A. Zaragoza, Paul E. Lee & Shana O. Kelley

  2. Department of Electrical and Computer Engineering, Faculty of Engineering, University of Toronto, Toronto, Canada

    Edward H. Sargent

  3. Department of Biochemistry, Faculty of Medicine, University of Toronto, Toronto, Canada

    Shana O. Kelley

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  1. Jagotamoy Das
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  2. Kristin B. Cederquist
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Contributions

S.O.K. and J.D. conceived the experiments. J.D., K.B.C., A.Z. and P.L. performed the experiments. J.D., K.B.C., A.Z., P.L., E.H.S. and S.O.K. discussed the results. J.D., K.B.C., E.H.S. and S.O.K. co-wrote and edited the manuscript.

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Correspondence to Shana O. Kelley.

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Das, J., Cederquist, K., Zaragoza, A. et al. An ultrasensitive universal detector based on neutralizer displacement. Nature Chem 4, 642–648 (2012). https://doi.org/10.1038/nchem.1367

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