Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 1391, Issue -, Pages 60-71Publisher
ELSEVIER
DOI: 10.1016/j.chroma.2015.02.070
Keywords
alpha-Gliadin; Celiac disease; LC-MRM/MS; T-cell stimulatory epitopes; Wheat
Funding
- European Regional Development Fund
- Province of Gelderland and Overijssel
- Dutch Ministry of Economic Affairs through the DLO program 'Plant and Animal for Human Health' [KB-05-001-019-PRI]
- Healthy and safe food in chain perspective [KB-15-001-016]
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Celiac disease (CD) is a food-related disease caused by certain gluten peptides containing T-cell stimulating epitopes from wheat, rye, and barley. CD-patients have to maintain a gluten-free diet and are therefore dependent on reliable testing and labeling of gluten-free products. So far, the R5-ELISA is the approved method to detect if food products can be labeled gluten-free. Because the R5-ELISA detects gluten in general, there is a demand for an improved detection method that quantifies specifically CD-epitopes. Therefore, we developed a new method for detection and quantification of CD-epitopes, based on liquid chromatography (LC) coupled to mass spectrometry (MS) in multiple reaction monitoring (MRM) mode. This method enables targeted label free comparative analysis of the gluten proteins present in different wheat varieties and species, and in wheat-based food products. We have tested our method by analyzing several wheat varieties that vary in CD-epitope content, as was shown before using immunoblotting and specific monoclonal antibodies. The results showed that a modern bread wheat variety Toronto contained the highest amounts of CD itninunogenic peptides compared with the older bread wheat variety Minaret and the tetraploid wheat variety Dibillik Sinde. Our developed method can detect quantitatively and simultaneously multiple specific CD-epitopes in a high throughput manner. (C) 2015 Elsevier B.V. All rights reserved.
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