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Focal molography is a new method for the in situ analysis of molecular interactions in biological samples

An Author Correction to this article was published on 11 February 2021

This article has been updated

Abstract

Focal molography is a next-generation biosensor that visualizes specific biomolecular interactions in real time. It transduces affinity modulation on the sensor surface into refractive index modulation caused by target molecules that are bound to a precisely assembled nanopattern of molecular recognition sites, termed the ‘mologram’. The mologram is designed so that laser light is scattered at specifically bound molecules, generating a strong signal in the focus of the mologram via constructive interference, while scattering at nonspecifically bound molecules does not contribute to the effect. We present the realization of molograms on a chip by submicrometre near-field reactive immersion lithography on a light-sensitive monolithic graft copolymer layer. We demonstrate the selective and sensitive detection of biomolecules, which bind to the recognition sites of the mologram in various complex biological samples. This allows the label-free analysis of non-covalent interactions in complex biological samples, without a need for extensive sample preparation, and enables novel time- and cost-saving ways of performing and developing immunoassays for diagnostic tests.

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Figure 1: Schematic illustration of the fundamental components of focal molography and the submicrometre near-field lithographic process used in molography.
Figure 2: Comparison of sandwich immunoassay performed by molography and OWLS (diffractometric versus refractometric biosensor).
Figure 3: Determination of assay specific affinity for antibody binding to β-amyloid peptide.
Figure 4: Real-time detection of a therapeutic antibody in human plasma.
Figure 5: In situ measurement of secreted immunoglobulins in a hybridoma cell culture.

Change history

  • 25 October 2017

    In the version of this Article originally published, the illumination pattern below the phase mask was incorrectly positioned in Fig. 1b (ii) and Zeptosens was misspelled in two instances in Methods. These errors have been corrected in all versions of the Article.

  • 11 February 2021

    A Correction to this paper has been published: https://doi.org/10.1038/s41565-021-00863-x

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Acknowledgements

The authors thank V. Guzenko (ETH) and W. Arens (IMT) for technical support in fabrication of the phase mask, S. Tosatti (SuSoS) and S. Zürcher (SuSoS) for consulting regarding surface science questions and production of the copolymer, A. Nichtl (Roche Diagnostics) for support with various reagents, H.P. Herzig (EPFL) and A. Naqavi (EPFL) for support with numerical simulations and J. Hehl and T. Schwarz (ScopeM/ETH) for STED support. For designing and fabricating numerous hardware components, the authors thank T. Kissling, R. Rietmann (Roche) and S. Wheeler (ETH). The authors also thank A. Lieb for support with ZeptoReader-related issues. The authors thank the following for discussions on various aspects of the project: M. Hennig, K. Mueller, M. Lauer, A. Rufer, G. Dernick, M. Marcinowski, M. Essenpreis, M. Hein, O. Gutmann, A. Drechsler, M. Glauser, N. Milicevic, J. Spinke, A. Maurer, C. Patsch, C. Cusulin, J. Fingerle, R. Staack and A. Poehler. The authors acknowledge the Roche Postdoc Fellowship (RPF) Program, ETH Zurich and the NCCR Molecular Systems Engineering for funding.

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Experiments were designed by V.G., K.-P.S., D.H., T.L., J.V. and C.F. C.F. performed the calculations for the phase mask and all other optical components. Molographic experiments were performed by V.G. A.F. performed the numerical simulations and wrote the evaluation software with support from J.V. and C.F. All authors read and approved the manuscript for submission.

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Correspondence to Janos Vörös or Christof Fattinger.

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Gatterdam, V., Frutiger, A., Stengele, KP. et al. Focal molography is a new method for the in situ analysis of molecular interactions in biological samples. Nature Nanotech 12, 1089–1095 (2017). https://doi.org/10.1038/nnano.2017.168

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