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Antigen-Specific Human Monoclonal Antibodies from Transgenic Mice

  • Susana Magadán Mompó
  • África González-Fernández
Part of the Methods in Molecular Biology book series (MIMB, volume 1060)

Abstract

Due to the difficulties found when generating fully human monoclonal antibodies (mAbs) by the traditional method, several efforts have attempted to overcome these problems, with varying levels of success. One approach has been the development of transgenic mice carrying immunoglobulin (Ig) genes in germ line configuration. The engineered mouse genome can undergo productive rearrangement in the B cell population, with the generation of mouse B lymphocytes expressing human Ig (hIg) chains. To avoid the expression of mouse heavy or light chains, the endogenous mouse Ig (mIg) loci must be silenced by gene-targeting techniques. Subsequently, to obtain antigen-specific mAbs, conventional immunization protocols can be followed and the mAb technique used (fusion of activated B cells with mouse myeloma cells, screening, cloning, freezing, and testing) with these animals expressing human Ig genes. This chapter describes the type of transgenic knockout mice generated for various research groups, provides examples of human mAbs developed by research groups and companies, and includes protocols of immunization, generation, production, and purification of human mAbs from such mice. In addition, it also addresses the problems detected, and includes some of the methods that can be used to analyze functional activities with human mAbs.

Key words

Human monoclonal antibodies Transgenic mice Immunoglobulin transgenes Knockout mice Transloci bearing human Ig genes Gene targeting YAC-based human Ig transloci 

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Copyright information

© Springer Science+Business Media, LLC 2014

Authors and Affiliations

  • Susana Magadán Mompó
    • 1
    • 2
  • África González-Fernández
    • 3
    • 4
  1. 1.Immunology, Biomedical Research Center (CINBIO)VigoSpain
  2. 2.Institute of Biomedical Research (IBIV)Universidad de VigoVigoSpain
  3. 3.Directora del Centro de Investigación Biomédica (CINBIO)VigoSpain
  4. 4.Catedrática de Inmunología Universidad de VigoVigoSpain

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