1. Combination of two epitope identification techniques enables the rational design of soy allergen Gly m 4 mutants
- Author
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Havenith, Heide, Kern, Karolin, Rautenberger, Paul, Spiegel, Holger, Szardenings, Michael, Ueberham, Elke, Lehmann, Jörg, Buntru, Matthias, Vogel, Simon, Treudler, Regina, Fischer, Rainer, Schillberg, Stefan, and Publica
- Subjects
epitope mapping ,soy allergy ,peptide microarray ,bet v 1 homolog ,phage display - 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
- Published
- 2017