Isolation of Natural Anti-FcεRIα Autoantibodies from Healthy Donors.

Vogel, Monique; Horn, Michael (2017). Isolation of Natural Anti-FcεRIα Autoantibodies from Healthy Donors. In: Natural Antibodies. Methods in molecular biology: Vol. 1643 (pp. 5-22). Springer 10.1007/978-1-4939-7180-0_2

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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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