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Antibody Engineering pp 195–209Cite as

Generation of Human Single Domain Antibody Repertoires by Kunkel Mutagenesis

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 907))

Abstract

Human antibody single domains are a promising new class of antibody fragments. Here we describe methods for the cloning of human VH and VL genes into phage and phagemid vectors. Furthermore, we provide detailed protocols for the generation of single domain antibody libraries by Kunkel mutagenesis and the analysis of diversity by DNA sequencing and superantigen binding.

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Acknowledgments

The protocols are based on methods originally developed in Greg Winter’s group at the MRC Laboratory of Molecular Biology and were modified in our laboratory at the Garvan Institute. This work was funded by the Garvan Institute of Medical Research, the Australian National Health and Medical Council, the Australian Research Council, the Cancer Institute NSW, and the United Kingdom Medical Research Council.

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Authors and Affiliations

  1. Garvan Institute of Medical Research, Darlinghurst/Sydney, NSW, Australia

    Romain Rouet, Kip Dudgeon & Daniel Christ

Authors
  1. Romain Rouet
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  2. Kip Dudgeon
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  3. Daniel Christ
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Corresponding author

Correspondence to Daniel Christ .

Editor information

Editors and Affiliations

  1. Antibody Therapeutics & Immunotargeting, INSERM U624, Marseille CEDEX 09, France

    Patrick Chames

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Rouet, R., Dudgeon, K., Christ, D. (2012). Generation of Human Single Domain Antibody Repertoires by Kunkel Mutagenesis. In: Chames, P. (eds) Antibody Engineering. Methods in Molecular Biology, vol 907. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-974-7_10

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  • DOI: https://doi.org/10.1007/978-1-61779-974-7_10

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-973-0

  • Online ISBN: 978-1-61779-974-7

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