title: Combination of two epitope identification techniques enables the rational design of soy allergen Gly m 4 mutants.
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.
authors.affiliation: Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany.
authors.forename: Heide
authors.initials: H
authors.lastname: Havenith
authors.affiliation: Fraunhofer Institute for Cell Therapy and Immunology IZI, Leipzig, Germany.
authors.forename: Karolin
authors.initials: K
authors.lastname: Kern
authors.affiliation: Fraunhofer Institute for Cell Therapy and Immunology IZI, Leipzig, Germany.
authors.forename: Paul
authors.initials: P
authors.lastname: Rautenberger
authors.affiliation: Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany.
authors.forename: Holger
authors.initials: H
authors.lastname: Spiegel
authors.affiliation: Fraunhofer Institute for Cell Therapy and Immunology IZI, Leipzig, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Szardenings
authors.affiliation: Fraunhofer Institute for Cell Therapy and Immunology IZI, Leipzig, Germany.
authors.forename: Elke
authors.initials: E
authors.lastname: Ueberham
authors.affiliation: Fraunhofer Institute for Cell Therapy and Immunology IZI, Leipzig, Germany.
authors.forename: Jörg
authors.initials: J
authors.lastname: Lehmann
authors.affiliation: Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Buntru
authors.affiliation: Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany.
authors.forename: Simon
authors.initials: S
authors.lastname: Vogel
authors.affiliation: Dermatology, Venereology and Allergology Clinic, University of Leipzig, Leipzig, Germany.
authors.forename: Regina
authors.initials: R
authors.lastname: Treudler
authors.affiliation: Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany.|RWTH Aachen University, Institute for Molecular Biotechnology, Aachen, Germany.
authors.forename: Rainer
authors.initials: R
authors.lastname: Fischer
authors.affiliation: Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany.|Institute for Phytopathology and Applied Zoology, Justus-Liebig University Giessen, Gießen, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Schillberg
mesh_terms: D000485:Allergens / Q000235:genetics* / Q000276:immunology; D018604:Epitope Mapping; D000939:Epitopes / Q000235:genetics* / Q000276:immunology*; D005512:Food Hypersensitivity / Q000235:genetics / Q000276:immunology; D009154:Mutation; D019151:Peptide Library; D010940:Plant Proteins / Q000235:genetics / Q000276:immunology; D013025:Glycine max / Q000235:genetics* / Q000276:immunology*
chemical_list: D000485:Allergens; D000939:Epitopes; D019151:Peptide Library; D010940:Plant Proteins
keywords: Bet v 1 homolog; Epitope mapping; Peptide microarray; Phage display; Soy allergy