1. Proteomic analysis of wheat proteins recognized by IgE antibodies of allergic patients.
- Author
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Sotkovský P, Hubálek M, Hernychová L, Novák P, Havranová M, Setinová I, Kitanovicová A, Fuchs M, Stulík J, and Tucková L
- Subjects
- Adolescent, Adult, Child, Electrophoresis, Gel, Two-Dimensional, Electrophoresis, Polyacrylamide Gel, Enzyme Inhibitors immunology, Enzyme Inhibitors metabolism, Enzyme-Linked Immunosorbent Assay, Flow Cytometry, Humans, Immunoblotting, Peptide Fragments analysis, Peptide Fragments metabolism, Plant Proteins immunology, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Triticum adverse effects, Triticum metabolism, Wheat Hypersensitivity diagnosis, alpha-Amylases antagonists & inhibitors, Allergens immunology, Immunoglobulin E blood, Plant Proteins metabolism, Proteome analysis, Triticum immunology, Wheat Hypersensitivity immunology
- Abstract
Wheat belongs to six major food allergens inducing IgE-mediated hypersensitivity reaction manifesting as cutaneous, gastrointestinal, and respiratory symptoms. Although cereals are a staple food item in most diets, only a few wheat proteins causing hypersensitivity have been identified. To characterize wheat allergens, salt-soluble wheat extracts were separated by 1-DE and 2-DE and IgE-binding proteins were detected by immunoblotting using sera of patients with allergy to ingested wheat. Proteins, frequently recognized by IgE on 2-DE were analyzed by MALDI-TOF and QTOF and their spectrum was completed by 1-DE and LCQ(DECA) nLC-MS/MS IT technique. Using all three techniques we identified 19 potential wheat allergens such as alpha-amylase inhibitors, beta-amylase, profilin, serpin, beta-D-glucan exohydrolase, and 27K protein. Employing newly developed ELISA, levels of IgE Abs against Sulamit wheat extract and alpha-amylase inhibitors type 1 and 3 were quantified and shown to be significantly elevated in sera of allergic patients compared to those of healthy controls. The level of IgE Abs against alpha-amylase inhibitor type 3 was lower, slightly above the cut-off value in the majority of patients' sera. Our findings contribute to the identification of wheat allergens aimed to increase the specificity of serum IgE and cell activation diagnostic assays.
- Published
- 2008
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