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Spatial Control of Biological Ligands on Surfaces Applied to T Cell Activation

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The Immune Synapse

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1584))

Abstract

In this chapter, we present techniques, based on molecular-scale nanofabrication and selective self-assembly, for the presentation of biomolecules of interest (ligands, receptors, etc.) on a surface with precise spatial control and arbitrary geometry at the single-molecule level. Metallic nanodot arrays are created on glass coverslips and are then used as anchors for the immobilization of biological ligands via thiol linking chemistry. The nanodot size is controlled by both lithography and metallization. The reagent concentration in self-assembly can be adjusted to ensure single-molecule occupancy for a given dot size. The surrounding glass is backfilled by a protein-repellent layer to prevent nonspecific adsorption. Moreover, bifunctional surfaces are created, whereby a second ligand is presented on the background, which is frequently a requirement for simulating complex cellular functions involving more than one key ligand. This platform serves as a novel and powerful tool for molecular and cellular biology, e.g., to study the fundamental mechanisms of receptor-mediated signaling.

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Acknowledgments

The authors thank Dr. M. Palma for guidance on nanoarray functionalization and Dr. Silvia Curado for coordinating our collaboration. This work was supported primarily by the National Science Foundation (NSF) under award no. CMMI-1300590 and by the National Institutes of Health (NIH) Common Fund Nanomedicine program grant PN2 EY016586. The authors are grateful to the Columbia Nano Initiative for providing cleanroom and other facilities used in this work.

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Authors and Affiliations

  1. Department of Mechanical Engineering, Columbia University, New York, USA

    Haogang Cai

  2. Kennedy Institute of Rheumatology, NDORMS, The University of Oxford, Oxford, UK

    David Depoil & Michael L. Dustin

  3. Department of Pathology, Skirball Institute, New York University School of Medicine, New York, USA

    James Muller & Michael L. Dustin

  4. Department of Biological Sciences, Columbia University, New York, USA

    Michael P. Sheetz

  5. National University of Singapore, Singapore, Singapore

    Michael P. Sheetz

  6. Department of Applied Physics and Applied Mathematics, Columbia University, 500 W 120th St, New York, NY, 10027, USA

    Shalom J. Wind

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  1. Haogang Cai
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  2. David Depoil
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  3. James Muller
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  4. Michael P. Sheetz
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  5. Michael L. Dustin
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Correspondence to Shalom J. Wind .

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Editors and Affiliations

  1. Department of Life sciences, University of Siena, Siena, Siena, Italy

    Cosima T. Baldari

  2. University of Oxford, Kennedy Institute of Rheumatology, Headington, Oxford, United Kingdom

    Michael L. Dustin

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Cai, H., Depoil, D., Muller, J., Sheetz, M.P., Dustin, M.L., Wind, S.J. (2017). Spatial Control of Biological Ligands on Surfaces Applied to T Cell Activation. In: Baldari, C., Dustin, M. (eds) The Immune Synapse. Methods in Molecular Biology, vol 1584. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6881-7_18

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  • DOI: https://doi.org/10.1007/978-1-4939-6881-7_18

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