Reference : Detection of wheat allergens using 2D Western blot and mass spectrometry
Scientific journals : Article
Human health sciences : Laboratory medicine & medical technology
Human health sciences : Public health, health care sciences & services
http://hdl.handle.net/2268/244821
Detection of wheat allergens using 2D Western blot and mass spectrometry
English
Courtois, Justine mailto [Université de Liège - ULiège > > I3-Hematology >]
BERTHOLET, Catherine mailto [Centre Hospitalier Universitaire de Liège - CHU > Unilab > Secteur commun prélèvements - dispa - labo central Corelab >]
Tollenaere, S. [CRIG, Liège, Belgium, HELMo, Liège, Belgium]
Van der Brempt, X. [Allergopôle, Clinique Saint-Luc, Bouge, Belgium]
Cavalier, Etienne mailto [Université de Liège - ULiège > Département de pharmacie > Chimie médicale >]
El Guendi, Sonia mailto [HELMO > > > >]
Gillard, N [CER Group > > > >]
Gadisseur, Romy [Université de Liège - ULiège > > CIRM >]
Quintig, Birgit [Helmo > > > >]
2020
Journal of Pharmaceutical and Biomedical Analysis
Elsevier B.V.
178
Yes (verified by ORBi)
International
07317085
[en] Mass spectrometry ; Western blot ; Wheat allergens ; Wheat allergy
[en] Background: Wheat allergy is relatively common and the associated clinical manifestations depend on the involved molecular allergens as well as on the way of exposure. Different symptoms have been described: wheat-dependent exercise-induced anaphylaxis (WDEIA), atopic dermatitis (AD) and pollen rhinitis (PR). Traditional diagnostic methods do not allow accurate molecular identification of the allergens that are essential for risk assessment and for the choice of the most adapted treatment. Methods: Standardized total protein extracts obtained from wheat seeds were separated by 2D electrophoresis. Twenty-one sera with high wheat-specific immunoglobulin E (sIgE) levels were classified into three patients groups based on their clinical profile. These sera were tested by Western blot on 2D separated standardized wheat protein extract and their sIgE sensitization profiles were compared. Results: Specific sensitization profiles were identified for each phenotype group. For WDEIA, protein spots around 37 kDa (pH 6-9) and 37–50 kDa (pH 5-6) were identified. For AD, spots were observed around 50 kDa (pH 9), 10 kDa (pH 9) and 20 to 75 kDa (pH3). For PR, specific spots were located around 90 kDa (pH 9). The mass spectrometry (UHPLC-MS/MS) analysis of these identified spots pointed out several potential interesting allergens: Tri a 26, Tri a bA, Tri a 34, Tri a tritin. Conclusions: The present study allowed the identification of different protein areas specific to these studied groups. The protein spots of interest were identified by UHPLC-MS/MS. It has been possible to establish a link between a specific symptomatology and the newly identified responsible allergens. © 2019 Elsevier B.V.
Fédération Wallonie-Bruxelles
http://hdl.handle.net/2268/244821
10.1016/j.jpba.2019.112907

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