Detection of wheat allergens using 2D Western blot and mass spectrometry

التفاصيل البيبلوغرافية
العنوان: Detection of wheat allergens using 2D Western blot and mass spectrometry
المؤلفون: S. El Guendi, X. Van der Brempt, Nathalie Gillard, Catherine Bertholet, Etienne Cavalier, Justine Courtois, Stéphanie Tollenaere, Romy Gadisseur, Birgit Quinting
المصدر: Journal of Pharmaceutical and Biomedical Analysis. 178:112907
بيانات النشر: Elsevier BV, 2020.
سنة النشر: 2020
مصطلحات موضوعية: Adult, Male, Adolescent, Blotting, Western, Clinical Biochemistry, Pharmaceutical Science, Wheat Hypersensitivity, Immunoglobulin E, 01 natural sciences, Analytical Chemistry, Young Adult, Western blot, Tandem Mass Spectrometry, Drug Discovery, medicine, Humans, Child, Chromatography, High Pressure Liquid, Spectroscopy, Sensitization, Two-dimensional gel electrophoresis, medicine.diagnostic_test, Spots, biology, 010405 organic chemistry, Chemistry, 010401 analytical chemistry, Atopic dermatitis, Allergens, Middle Aged, medicine.disease, Molecular biology, 0104 chemical sciences, medicine.anatomical_structure, Child, Preschool, biology.protein, Female, Wheat allergy, Anaphylaxis
الوصف: 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.
تدمد: 0731-7085
الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::e54cecbab5924097c28ac32b54663effTest
https://doi.org/10.1016/j.jpba.2019.112907Test
حقوق: CLOSED
رقم الانضمام: edsair.doi.dedup.....e54cecbab5924097c28ac32b54663eff
قاعدة البيانات: OpenAIRE