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Generating Conformation and Complex-Specific Synthetic Antibodies

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Book cover Synthetic Antibodies

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1575))

Abstract

Phage display is commonly used to identify and isolate binders from large combinatorial libraries. Here we present phage selection protocols enabling generation of synthetic antibodies capable of recognizing multiprotein complexes and conformational states. The procedure describes stages of the experiment design, optimization, and screening, as well as provides the framework for building downstream assays with an end goal of isolating bioactive antibodies for future therapeutic use. The methods described are also applicable to screening directly on cells and can be ported to other in vitro directed evolution systems utilizing non-immunoglobulin scaffolds.

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Acknowledgments

We thank Vincent Lu, Daniel King, and Svitlana Usatyuk for contributing to the development of protocols. This work was supported by National Institutes of Health Grants: U54-GM087519, U54-HG006436.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, The University of Chicago, KCBD Rm.3240G, 900 East 57th Street, Chicago, IL, 60637, USA

    Marcin Paduch & Anthony A. Kossiakoff

  2. Institute for Biophysical Dynamics, The University of Chicago, Chicago, IL, 60637, USA

    Anthony A. Kossiakoff

Authors
  1. Marcin Paduch
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  2. Anthony A. Kossiakoff
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Corresponding author

Correspondence to Marcin Paduch .

Editor information

Editors and Affiliations

  1. MorphoSys AG, Discovery Alliances & Technologies, Planegg, Germany

    Thomas Tiller

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Paduch, M., Kossiakoff, A.A. (2017). Generating Conformation and Complex-Specific Synthetic Antibodies. In: Tiller, T. (eds) Synthetic Antibodies. Methods in Molecular Biology, vol 1575. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6857-2_6

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  • DOI: https://doi.org/10.1007/978-1-4939-6857-2_6

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6855-8

  • Online ISBN: 978-1-4939-6857-2

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