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Production of Human or Humanized Antibodies in Mice

  • Brice Laffleur
  • Virginie Pascal
  • Christophe Sirac
  • Michel CognéEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 901)

Abstract

Mice are widely available laboratory animals that can easily be used for the production of antibodies against a broad range of antigens, using well-defined immunization protocols. Such an approach allows optimal in vivo affinity maturation of the humoral response. In addition, high-affinity antibodies arising in this context can readily be further characterized and produced as monoclonals after immortalizing and selecting specific antibody-producing cells through hybridoma derivation. Using such conventional strategies combined with mice that are either genetically engineered to carry humanized immunoglobulin (Ig) genes or engrafted with a human immune system, it is thus easy to obtain and immortalize clones that produce either fully human Ig or antibodies associating variable (V) domains with selected antigen specificities to customized human-like constant regions, with defined effector functions. In some instances, where there is a need for in vivo functional assays of a single antibody with a known specificity, it might be of interest to transiently express that gene in mice by in vivo gene transfer. This approach allows a rapid functional assay. More commonly, mice are used to obtain a diversified repertoire of antibody specificities after immunization by producing antibody molecules in the mouse B cell lineage from mouse strains with transgene Ig genes which are of human, humanized, or chimeric origin. After in vivo maturation of the immune response, this will lead to the secretion of antibodies with optimized antigen binding sites, associated to the desired human constant domains. This chapter focuses on two simple methods: (1) to obtain such humanized Ig mice and (2) to transiently express a human Ig gene in mice using hydrodynamics-based transfection.

Key words

B cells ES cells Genetic engineered mice Homologous recombination Humanized mice Humanized antibodies Hydrodynamics-based transfection Immunoglobulin 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Brice Laffleur
    • 1
  • Virginie Pascal
    • 1
  • Christophe Sirac
    • 1
  • Michel Cogné
    • 1
    Email author
  1. 1.CNRS UMR6101, Contrôle des Réponses Immunes B et LymphoproliférationsUniversité de LimogesLimogesFrance

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