|
|
Food allergy to wheat flour proteins
Šotkovský, Petr ; Tučková, Ludmila (advisor) ; Panzner, Petr (referee) ; Prokešová, Ludmila (referee)
THESIS SUMMARY Food allergy is one of the frequent disorders and its incidence in paediatric as well as adult population is continuously rising, having doubled in the last two decades. Although wheat belongs to major food allergens and is a staple food in most diets, we have only little knowledge of wheat proteins causing IgE mediated hypersensitivity reaction. Diagnostic approaches of food allergy to wheat have a high sensitivity, but low specificity. Poor predictability and specificity may be associated with the insufficient purity of wheat extracts used in sIgE assays or with the lack of major allergens in these extracts. In the first step, we characterized 19 potential allergens recognised by IgE Abs of allergic patients, using proteomic techniques (1-DE, 2-DE, MALDI-TOF, QTOF and LCQDECA nLC-MS/MS ion trap technique). We identified these IgE-binding molecules such as: α-amylase inhibitors, β-amylase, profilin, serpin, β-D-glucan exohydrolase and 27K protein. To quantify sIgE in patient's sera we developed ELISA using the whole wheat extract and two commercially available α-amylase inhibitors. Second, we developed a procedure that allows isolation of wheat allergens from natural sources using Rotofor cell and HPLC. Twenty-seven potential wheat allergens have been successfully identified; of these, the...
|
guest ::
