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Construction of Naïve and Immune Human Fab Phage Display Library

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Phage Display

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2702))

Abstract

Phage display has been applied successfully for the rapid isolation of monoclonal antibodies against various targets including infectious diseases, autoantigens, cancer markers, and even small molecules. The main component in any phage display experiment is the availability of an antibody library to carry out the selection process of target-specific antibodies through an iterative process termed as biopanning. To generate human antibody libraries, the antibody repertoire can be obtained from human peripheral blood mononuclear cell (PBMC) or directly from cell-sorted B-cell populations. The choice of antibody isotype is dictated by the nature of the library. Naïve libraries would utilize IgM repertoires, whereas the IgG repertoire is commonly used for immune libraries. Antibody genes are amplified through polymerase chain reaction (PCR) and paired in a combinatorial fashion to expand the diversity of the cloned library repertoire. The protocol here describes the use of a two-step cloning method that can be applied for the construction of either a naïve or immune human antibody library in Fab format followed by the subsequent panning.

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Acknowledgments

This work was supported by a Universiti Sains Malaysia, Special (Matching) Short-Term Grant with Project No: 304/CIPPM/6315708.

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Authors and Affiliations

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

    Jing Yi Lai & Theam Soon Lim

Authors
  1. Jing Yi Lai
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  2. Theam Soon Lim
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Corresponding author

Correspondence to Theam Soon Lim .

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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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Lai, J.Y., Lim, T.S. (2023). Construction of Naïve and Immune Human Fab Phage Display Library. 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_3

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  • DOI: https://doi.org/10.1007/978-1-0716-3381-6_3

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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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