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.

• MeSH
• 2D gelová elektroforéza MeSH
• alergeny imunologie   MeSH
• alergie na pšenici diagnóza   imunologie   MeSH
• alfa-amylasy antagonisté a inhibitory   MeSH
• dítě MeSH
• dospělí MeSH
• elektroforéza v polyakrylamidovém gelu MeSH
• ELISA MeSH
• financování organizované MeSH
• imunoblotting MeSH
• imunoglobulin E krev   MeSH
• inhibitory enzymů imunologie   metabolismus   MeSH
• lidé MeSH
• mladiství MeSH
• peptidové fragmenty analýza   metabolismus   MeSH
• průtoková cytometrie MeSH
• pšenice imunologie   metabolismus   škodlivé účinky   MeSH
• rostlinné proteiny imunologie   metabolismus   MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• mladiství MeSH

• Publikační typ
• abstrakt z konference MeSH