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Isolation of Natural Anti-FcεRIα Autoantibodies from Healthy Donors

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Natural Antibodies

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1643))

Abstract

Natural antibodies are defined as antibodies detected in a healthy individual without active immunization. These antibodies are specific for exoantigens, as well as for autoantigens, mostly without any pathogenic role. Most of the studies conducted with natural (auto-) antibodies have been performed using affinity purified antibodies from individual sera or polyclonal Ig-preparations such as Intravenous Ig (IVIg). For in-depth analysis of such autoantibodies affinity-purified Ig-preparations from healthy individuals are of no use, as they are oligoclonal or polyclonal. Thus, there is a need of human monoclonal autoantibodies. Human monoclonal autoantibodies can be produced from B cells isolated from humans; however, this requires the screening of a large number of antibodies to identify one among them specific to an antigen. Using the phage display technology we generated such autoantibodies against the alpha subunit of the high-affinity IgE receptor (FcεRIα). Here we describe the step-by-step protocol for the generation of such libraries and the isolation of autoantibodies by affinity panning.

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Author information

Authors and Affiliations

  1. University Clinic of Rheumatology, Immunology and Allergology, Inselspital, University Hospital of Bern, CH-3010, Bern, Switzerland

    Monique Vogel

  2. University Institute of Clinical Chemistry and Center of Laboratory Medicine, Inselspital, University Hospital of Bern, CH-3010, Bern, Switzerland

    Michael P. Horn

Authors
  1. Monique Vogel
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  2. Michael P. Horn
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Corresponding author

Correspondence to Monique Vogel .

Editor information

Editors and Affiliations

  1. Centre de Recherche des Cordeliers, INSERM, PARIS, Paris, France

    Srinivas V. Kaveri

  2. Centre de Recherche des Cordeliers, INSERM, PARIS, Paris, France

    Jagadeesh Bayry

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Vogel, M., Horn, M.P. (2017). Isolation of Natural Anti-FcεRIα Autoantibodies from Healthy Donors. In: Kaveri, S., Bayry, J. (eds) Natural Antibodies. Methods in Molecular Biology, vol 1643. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7180-0_2

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  • DOI: https://doi.org/10.1007/978-1-4939-7180-0_2

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7179-4

  • Online ISBN: 978-1-4939-7180-0

  • eBook Packages: Springer Protocols

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