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Background: Detailed IgE‐binding epitope analysis is a key requirement for the understanding and development of diagnostic and therapeutic agents to address food allergies. Methods: We combined 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 v1. Results: Three epitopes were 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 was reduced by up to 63% compared to the wild‐type allergen. Basophil stimulation experiments using RBL‐SX38 cells loaded with patient IgE showed a decreased stimulation from 25% for the wild‐type Gly m 4 to 13% for one mutant. Conclusion: 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.NEXT ARTICLE
Food is any substance consumed to provide nutritional support for the body. It is usually of plant or animal origin, and contains essential nutrients, such as carbohydrates, fats, proteins, vitamins, or minerals. The substance is ingested by an organism ...
The term allergy is used to describe a response, within the body, to a substance, which is not necessarily harmful in itself, but results in an immune response and a reaction that causes symptoms and disease in a predisposed person, which in turn can cau...
A DNA microarray or biochip is a collection of microscopic DNA spots attached to a solid surface used to measure the expression levels of large numbers of genes simultaneously or to genotype multiple regions of a genome.