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Combination of two epitope identification techniques enables the rational design of soy allergen Gly m 4 mutants
Authors:Heide Havenith  Karolin Kern  Paul Rautenberger  Holger Spiegel  Michael Szardenings  Elke Ueberham  Jörg Lehmann  Matthias Buntru  Simon Vogel  Regina Treudler  Rainer Fischer  Stefan Schillberg
Institution:1. Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany;2. Fraunhofer Institute for Cell Therapy and Immunology IZI, Leipzig, Germany;3. Dermatology, Venereology and Allergology Clinic, University of Leipzig, Leipzig, Germany;4. RWTH Aachen University, Institute for Molecular Biotechnology, Aachen, Germany;5. Institute for Phytopathology and Applied Zoology, Justus‐Liebig University Giessen, Gie?en, Germany
Abstract:Detailed IgE‐binding epitope analysis is a key requirement for the understanding and development of diagnostic and therapeutic agents to address food allergies. An IgE‐specific linear peptide microarray with random phage peptide display for the high‐resolution mapping of IgE‐binding epitopes of the major soybean allergen Gly m 4, which is a homologue to the birch pollen allergen Bet v 1 is combined. Three epitopes are identified and mapped to a resolution of four key amino acids, allowing the rational design and the production of three Gly m 4 mutants with the aim to abolish or reduce the binding of epitope‐specific IgE. In ELISA, the binding of the mutant allergens to polyclonal rabbit‐anti Gly m 4 serum as well as IgE purified from Gly m 4‐reactive soybean allergy patient sera is reduced by up to 63% compared to the wild‐type allergen. Basophil stimulation experiments using RBL‐SX38 cells loaded with patient IgE are showed a decreased stimulation from 25% for the wild‐type Gly m 4 to 13% for one mutant. The presented approach demonstrates the feasibility of precise mapping of allergy‐related IgE‐binding epitopes, allowing the rational design of less allergenic mutants as potential therapeutic agents.
Keywords:Bet v 1 homolog  Epitope mapping  Peptide microarray  Phage display  Soy allergy
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