Chapter title |
Isolation of Natural Anti-FcεRIα Autoantibodies from Healthy Donors
|
---|---|
Chapter number | 2 |
Book title |
Natural Antibodies
|
Published in |
Methods in molecular biology, July 2017
|
DOI | 10.1007/978-1-4939-7180-0_2 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7179-4, 978-1-4939-7180-0
|
Authors |
Monique Vogel, Michael P. Horn, Vogel, Monique, Horn, Michael P. |
Editors |
Srinivas V. Kaveri, Jagadeesh Bayry |
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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