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Single Domain Antibodies pp 327–353Cite as

Selecting and Purifying Autonomous Human Variable Heavy (VH) Domains

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 911))

Abstract

Antibodies are invaluable macromolecules effectively utilized as detection reagents and therapeutics. Traditionally, researchers have relied upon the entire immunoglobulin molecule, however advances in protein engineering have ushered the use of antibody fragments as equally important biological tools such that at present, the downstream application generally dictates the antibody format employed. We provide herein robust and proven protocols for the isolation of autonomous human antibody variable heavy domains (VH). The strategy utilizes combinatorial phage-displayed libraries targeting human VH domain positions previously shown to promote autonomous behavior, and selection against a specified antigen. Subsequently, autonomous VH domains are characterized and chosen using standard biophysical methods.

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Author information

Authors and Affiliations

  1. Terrence Donnelly Center for Cellular and Biomolecular Research and Banting and Best Department of Medical Research, University of Toronto, Toronto, ON, Canada

    Raffi Tonikian & Sachdev S. Sidhu

  2. Department of Protein Engineering, Biogen Idec, Cambridge, MA, USA

    Raffi Tonikian

  3. Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada

    Sachdev S. Sidhu

Authors
  1. Raffi Tonikian
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  2. Sachdev S. Sidhu
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Corresponding author

Correspondence to Sachdev S. Sidhu .

Editor information

Editors and Affiliations

  1. , Molecular & Cellular Interactions, Vrije Universiteit Brussel, Pleinlaan 2, Brussels, 1050, Belgium

    Dirk Saerens

  2. , Molecular & Cellular Interactions, Vrije Universiteit Brussel, Pleinlaan 2, Brussels, 1050, Belgium

    Serge Muyldermans

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Tonikian, R., Sidhu, S.S. (2012). Selecting and Purifying Autonomous Human Variable Heavy (VH) Domains. In: Saerens, D., Muyldermans, S. (eds) Single Domain Antibodies. Methods in Molecular Biology, vol 911. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-968-6_20

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  • DOI: https://doi.org/10.1007/978-1-61779-968-6_20

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-967-9

  • Online ISBN: 978-1-61779-968-6

  • eBook Packages: Springer Protocols

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