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Stochastic sensing of proteins with receptor-modified solid-state nanopores

Nature Nanotechnology volume 7, pages 257–263 (2012)Cite this article

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Abstract

Solid-state nanopores are capable of the label-free analysis of single molecules. It is possible to add biochemical selectivity by anchoring a molecular receptor inside the nanopore, but it is difficult to maintain single-molecule sensitivity in these modified nanopores. Here, we show that metallized silicon nitride nanopores chemically modified with nitrilotriacetic acid receptors can be used for the stochastic sensing of proteins. The reversible binding and unbinding of the proteins to the receptors is observed in real time, and the interaction parameters are statistically analysed from single-molecule binding events. To demonstrate the versatile nature of this approach, we detect His-tagged proteins and discriminate between the subclasses of rodent IgG antibodies.

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Figure 1: Chemical modification of metallized solid-state nanopores and stochastic sensing of proteins.
Figure 2: Single-molecule binding kinetics of His6-tagged protein A/G/L to a bisNTA modified nanopore.
Figure 3: Quantitation of dissociation rates and influence of receptor position.
Figure 4: Influence of receptor multivalence on binding stability.
Figure 5: Antibody-selective nanopore.

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Acknowledgements

The authors thank G. Abstreiter for his support, and M. Firnkes, D. Pedone, A. Kleefen and M. Langecker for discussions. This work was supported by the German Research Foundation DFG (SFB 863 and SFB 807), the TUM Institute for Advanced Study, the Federal Ministry of Education and Research BMBF (0312031/0312034), and the Clusters of Excellence Nanosystems Initiative Munich and Macromolecular Complexes (at the Goethe-University Frankfurt).

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Authors and Affiliations

  1. Walter Schottky Institute & Institute for Advanced Study, Technische Universität München, Am Coulombwall 4, Garching, 85748, Germany

    Ruoshan Wei & Ulrich Rant

  2. Institute of Biochemistry, Biocenter, Center for Membrane Proteomics (CMP), and Cluster of Excellence Frankfurt (CEF) Macromolecular Complexes, Goethe-University Frankfurt, Max-von-Laue-Strasse 9, Frankfurt am Main, 60438, Germany

    Volker Gatterdam, Ralph Wieneke & Robert Tampé

  3. Correspondence regarding thiols and surface chemistry should be addressed to R.T.,

    Robert Tampé

  4. Correspondence and requests for materials should be addressed to U.R.,

    Ulrich Rant

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Contributions

U.R., R.We. and R.T. devised the research. R.We. prepared the pores and performed experiments. V.G. and R.Wi. synthesized the NTA compounds. R.We., U.R. and R.T. analysed the data. R.We. and U.R. wrote the paper.

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Correspondence to Robert Tampé or Ulrich Rant.

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Wei, R., Gatterdam, V., Wieneke, R. et al. Stochastic sensing of proteins with receptor-modified solid-state nanopores. Nature Nanotech 7, 257–263 (2012). https://doi.org/10.1038/nnano.2012.24

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