title: Label free targeted detection and quantification of celiac disease immunogenic epitopes by mass spectrometry.
abstract: 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 immunogenic 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.
authors: Wageningen UR, Plant Research International, PO Box 16, 6700 AA Wageningen, The Netherlands. Electronic address: hetty.busink@wur.nl.
authors: Hetty C
authors: HC
authors: van den Broeck
authors: Wageningen UR, Plant Research International, PO Box 16, 6700 AA Wageningen, The Netherlands.
authors: Jan H G
authors: JH
authors: Cordewener
authors: RIKILT Wageningen UR, PO Box 230, 6700 AE Wageningen, The Netherlands.
authors: Merel A
authors: MA
authors: Nessen
authors: Wageningen UR, Plant Research International, PO Box 16, 6700 AA Wageningen, The Netherlands.
authors: Antoine H P
authors: AH
authors: America
authors: Wageningen UR, Plant Research International, PO Box 16, 6700 AA Wageningen, The Netherlands.
authors: Ingrid M
authors: IM
authors: van der Meer
mesh_terms: D002446:Celiac Disease / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000032:analysis / Q000276:immunology; D005983:Glutens / Q000032:analysis / Q000276:immunology; D006801:Humans; D013058:Mass Spectrometry; D010455:Peptides / Q000032:analysis / Q000276:immunology; D014908:Triticum / Q000737:chemistry / Q000276:immunology
chemical_list: D018984:Epitopes, T-Lymphocyte; D010455:Peptides; D005983:Glutens
keywords: Celiac disease; LC-MRM/MS; T-cell stimulatory epitopes; Wheat; α-Gliadin