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Antigen-Specific Human Monoclonal Antibodies from Transgenic Mice

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Human Monoclonal Antibodies

Part of the book series: Methods in Molecular Biology ((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.

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Author information

Authors and Affiliations

  1. Immunology, Biomedical Research Center (CINBIO), Vigo, Spain

    Susana Magadán Mompó

  2. Institute of Biomedical Research (IBIV), Universidad de Vigo, Vigo, Spain

    Susana Magadán Mompó

  3. Directora del Centro de Investigación Biomédica (CINBIO), Vigo, Pontevedra, Spain

    África González-Fernández

  4. Catedrática de Inmunología Universidad de Vigo, Vigo, Pontevedra, Spain

    África González-Fernández

Authors
  1. Susana Magadán Mompó
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  2. África González-Fernández
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Editors and Affiliations

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

    Michael Steinitz

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Mompó, S.M., González-Fernández, Á. (2014). Antigen-Specific Human Monoclonal Antibodies from Transgenic Mice. In: Steinitz, M. (eds) Human Monoclonal Antibodies. Methods in Molecular Biology, vol 1060. Humana, Totowa, NJ. https://doi.org/10.1007/978-1-62703-586-6_13

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  • DOI: https://doi.org/10.1007/978-1-62703-586-6_13

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  • Publisher Name: Humana, Totowa, NJ

  • Print ISBN: 978-1-62703-585-9

  • Online ISBN: 978-1-62703-586-6

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