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Antibody Fragments Humanization: Beginning with the End in Mind

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Human Monoclonal Antibodies

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1904))

Abstract

Molecular engineering has made possible to reformat monoclonal antibodies into smaller antigen-binding structures like scFvs, diabodies, Fabs with new potential in vivo applications because they do not induce Fc-mediated functions. However, most of these molecules are from rodent origin. As a consequence, they are immunogenic and approval for administration to humans requires prior humanization. Today, there is no well-identified strategy to create recombinant humanized antibody V-domains that preserve the antigen-binding characteristics of the parental antibody associated with high stability and solubility. Here, we propose a strategy that consists in grafting CDRs onto properly chosen human antibody frameworks in order to reduce immunogenicity. A flowchart indicates the way to proceed in order to introduce an internal affinity purification tag while structural refinements are proposed to maintain antigen-binding characteristics. The best humanized candidates are identified through selection steps including in silico analysis, research scale production followed by early functional evaluation, purification assays, aggregation, and stability assessment.

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Author information

Authors and Affiliations

  1. UMR Université-INRA ISP 1282, BioMAP, Université de Tours, Tours, France

    Nicolas Aubrey

  2. School of Pharmacy, Paris-Sud University, Châtenay-Malabry, France

    Philippe Billiald

Authors
  1. Nicolas Aubrey
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  2. Philippe Billiald
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Corresponding author

Correspondence to Philippe Billiald .

Editor information

Editors and Affiliations

  1. Department of Pathology, The Hebrew University, Hadassah Medical School, Jerusalem, Israel

    Michael Steinitz

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Aubrey, N., Billiald, P. (2019). Antibody Fragments Humanization: Beginning with the End in Mind. In: Steinitz, M. (eds) Human Monoclonal Antibodies. Methods in Molecular Biology, vol 1904. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8958-4_10

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  • DOI: https://doi.org/10.1007/978-1-4939-8958-4_10

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8957-7

  • Online ISBN: 978-1-4939-8958-4

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