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Preparation of Chromobodies for the Detection of Cell Surface Epitopes

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Mammalian Synthetic Systems

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2774))

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Abstract

Chromobodies are nanobodies genetically fused to fluorescent proteins, which were developed to visualize endogenous intracellular antigens. These versatile bioimaging nanotools can also be used to detect cell surface epitopes, and we describe here how we use them as an alternative to conjugated antibodies. This way, we routinely test the binding efficiency of nanobodies for their cognate cell surface antigens, before integrating them as sensing domains into complex synthetic receptor architectures.

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Acknowledgements

This work was supported by the School of Biological Sciences at the University of Edinburgh. Flow cytometry data were generated within the Flow Cytometry and Cell Sorting Facility in King’s Buildings at the University of Edinburgh, with the help of Dr Martin Waterfall. Microscopy images were acquired on a microscope funded by the School of Biological Sciences, the Institute of Quantitative Biology, Biochemistry, and Biotechnology, and the UK Centre for Mammalian Synthetic Biology.

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Authors and Affiliations

  1. Centre for Engineering Biology, School of Biological Sciences, The University of Edinburgh, Edinburgh, United Kingdom

    Ugne Baronaite & Elise Cachat

  2. Institute for Quantitative Biology, Biochemistry and Biotechnology, School of Biological Sciences, The University of Edinburgh, Edinburgh, United Kingdom

    Ugne Baronaite & Elise Cachat

Authors
  1. Ugne Baronaite
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  2. Elise Cachat
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Correspondence to Elise Cachat .

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Editors and Affiliations

  1. Department of Chemical Engineering and Imperial College Centre for Synthetic Biology, Imperial College London, London, UK

    Francesca Ceroni

  2. Department of Chemical Engineering and Imperial College Centre for Synthetic Biology, Imperial College London, London, UK

    Karen Polizzi

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Baronaite, U., Cachat, E. (2024). Preparation of Chromobodies for the Detection of Cell Surface Epitopes. In: Ceroni, F., Polizzi, K. (eds) Mammalian Synthetic Systems. Methods in Molecular Biology, vol 2774. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3718-0_20

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  • DOI: https://doi.org/10.1007/978-1-0716-3718-0_20

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3717-3

  • Online ISBN: 978-1-0716-3718-0

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