Isolation of Natural Anti-FcεRIα Autoantibodies from Healthy Donors

  • Monique VogelEmail author
  • Michael P. Horn
Part of the Methods in Molecular Biology book series (MIMB, volume 1643)


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.

Key words

Natural antibodies Anti-FcεRIα autoantibodies IVIg Phage display technology Isolation 


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© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.University Clinic of Rheumatology, Immunology and Allergology, Inselspital, University Hospital of BernBernSwitzerland
  2. 2.University Institute of Clinical Chemistry and Center of Laboratory Medicine, Inselspital, University Hospital of BernBernSwitzerland

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