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Construction of Synthetic Antibody Phage-Display Libraries

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Phage Display

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1701))

Abstract

Synthetic antibody libraries provide a vast resource of renewable antibody reagents that can rival or exceed those of natural antibodies and can be rapidly isolated through controlled in vitro selections. Use of highly optimized human frameworks enables the incorporation of defined diversity at positions that are most likely to contribute to antigen recognition. This protocol describes the construction of synthetic antibody libraries based on a single engineered human autonomous variable heavy domain scaffold with diversity in all three complementarity-determining regions. The resulting libraries can be used to generate recombinant domain antibodies for a wide range of protein antigens using phage display. Furthermore, analogous methods can be used to construct antibody libraries based on larger antibody fragments or second-generation libraries aimed to fine-tune antibody characteristics including affinity, specificity, and manufacturability. The procedures rely on standard reagents and equipment available in most molecular biology laboratories.

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Acknowledgments

Members of the Sidhu lab are acknowledged for input, particularly Alia Pavlenco and Wei Ye. This work was supported by funds from the Swedish Research Council (637-2013-468 to J.N.). We thank Frederic Fellouse for assistance with Fig. 3.

Author information

Authors and Affiliations

  1. Division of Protein Technology, School of Biotechnology, Royal Institute of Technology, Stockholm, Sweden

    Johan Nilvebrant

  2. Donnelly Centre for Cellular and Biomolecular Research, Banting and Best Department of Medical Research, University of Toronto, Room 816, 160 College Street, Toronto, ON, Canada, M5S 3E1

    Johan Nilvebrant & Sachdev S. Sidhu

Authors
  1. Johan Nilvebrant
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  2. Sachdev S. Sidhu
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Correspondence to Sachdev S. Sidhu .

Editor information

Editors and Affiliations

  1. Technische Universität Braunschweig, Braunschweig, Germany

    Michael Hust

  2. Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Minden, Penang, Malaysia

    Theam Soon Lim

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Nilvebrant, J., Sidhu, S.S. (2018). Construction of Synthetic Antibody Phage-Display Libraries. In: Hust, M., Lim, T. (eds) Phage Display. Methods in Molecular Biology, vol 1701. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7447-4_3

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  • DOI: https://doi.org/10.1007/978-1-4939-7447-4_3

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7446-7

  • Online ISBN: 978-1-4939-7447-4

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