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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 2702))

Abstract

Synthetic antibody libraries provide a vast resource of renewable antibody reagents that can rival natural antibodies and 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 targeting 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. We thank Frederic Fellouse for assistance with Fig. 3.

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Authors and Affiliations

  1. Division of Protein Engineering, School of Chemistry, Biotechnology and Health, Royal Institute of Technology, Stockholm, Sweden

    Kim Anh Giang & Johan Nilvebrant

  2. School of Pharmacy, University of Waterloo, Kitchener, ON, Canada

    Sachdev S. Sidhu

Authors
  1. Kim Anh Giang
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  2. Sachdev S. Sidhu
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  3. Johan Nilvebrant
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Corresponding author

Correspondence to Johan Nilvebrant .

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Editors and Affiliations

  1. Institut für Biochemie, Biotechnologie und Bioinformatik, Technische Universität Braunschweig, Braunschweig, Germany

    Michael Hust

  2. Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Penang, Malaysia

    Theam Soon Lim

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Giang, K.A., Sidhu, S.S., Nilvebrant, J. (2023). Construction of Synthetic Antibody Phage Display Libraries. In: Hust, M., Lim, T.S. (eds) Phage Display. Methods in Molecular Biology, vol 2702. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3381-6_4

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  • DOI: https://doi.org/10.1007/978-1-0716-3381-6_4

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3380-9

  • Online ISBN: 978-1-0716-3381-6

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