Mapping of the IgE and IgG4 Sequential Epitopes of Ovomucoid with a Peptide Microarray Immunoassay
| Autor: | Diego Gómez-Coronado, Inmaculada Cerecedo, S Terrados, Verónica de Dios, Belén de la Hoz, Javier Zamora, Maria Carmen Dieguez, Javier Martínez-Botas, Cristina Vlaicu |
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| Rok vydání: | 2012 |
| Předmět: |
Male
Adolescent Immunology Protein Array Analysis Ovomucin Immunoglobulin E medicine.disease_cause Epitope Epitopes Allergen medicine Humans Immunology and Allergy Child Egg Hypersensitivity biology medicine.diagnostic_test General Medicine medicine.disease Epitope mapping Child Preschool Immunoglobulin G Egg allergy Immunoassay biology.protein Female Peptide microarray Epitope Mapping |
| Zdroj: | International Archives of Allergy and Immunology. 161:11-20 |
| ISSN: | 1423-0097 1018-2438 |
| DOI: | 10.1159/000343040 |
| Popis: | Background: Ovomucoid (Gal d 1) has been demonstrated to be the most important allergen in IgE-mediated egg allergy. Peptide microarray analysis is a novel method that can provide useful information on the nature of specific allergens. Methods: A peptide microarray immunoassay was performed using a 15- and 20-amino acid (aa) library of overlapping peptides (3-offset) of the primary sequence of ovomucoid. Sera from 50 patients with IgE-mediated egg allergy and reactivity to ovomucoid, with more than 1 year of follow-up, and sera from 10 controls were tested. Peptides were considered major epitopes when the average weighted Z-score was greater than 3 and recognized by at least 20% of the patient’s sera. Specific IgE epitopes were established on the basis of the IgE/IgG4 Z-score ratio. Results: The IgE and IgG4 recognition pattern was similar in both sets of peptides, but the signal intensity was generally higher in the 20-aa set. Thirty-four percent of the patients did not recognize any IgE sequential peptide and 20% of the patients recognized more than 10 sequential peptides. We identified 3 major IgE B-cell epitopes in domains I and II of ovomucoid. IgE/IgG4 ratio analysis showed that peptides 1–2 (aa 4–20) and peptides 29–31 (aa 91–104) were specific IgE epitopes. Conclusion: By using peptide microarray immunoassay in egg-allergic patients, we established that 34% of the patients do not have any linear epitope recognized by IgE. Further studies are needed to determine the clinical relevance of this finding. |
| Databáze: | OpenAIRE |
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