Proteomics approaches to developing mass spectrometry methods for detection of gluten in gluten-free foods
Lead Research Organisation:
University of Manchester
Department Name: School of Biological Sciences
Abstract
Individuals with the gluten intolerance syndrome coeliac disease have to practice life-long avoidance of gluten-containing foods. As a consequence they rely on gluten-free foods which have to contain <20mg/Kg gluten. Currently immunoassay-based methods only detect a sub-group of gluten proteins and can give false positive results due to cross-reactivity with grass seeds proteins. The proposed project will give the student training in gluten protein chemistry, protein mass spectrometry and associated bioinformatics. The work is based in the Manchester Institute of Biotechnology with access to the state-of-the-art facilities. The project is sponsored by the Waters Corporation based at Wilmslow and will involve training periods in their laboratory. This will give the student the opportunity to translate their research into practical methods required by the food sector to support food allergen management and the development of effective methodology to verify "gluten-free" claims made on food products.
People |
ORCID iD |
| Clare Mills (Primary Supervisor) |
Publications
Bucchini L
(2019)
Health Claims and Food Labelling
Daly M
(2023)
A chromatographic and immunoprofiling approach to optimising workflows for extraction of gluten proteins from flour.
in Journal of chromatography. B, Analytical technologies in the biomedical and life sciences
Daly M
(2020)
Mapping Coeliac Toxic Motifs in the Prolamin Seed Storage Proteins of Barley, Rye, and Oats Using a Curated Sequence Database.
in Frontiers in nutrition
Daly ME
(2022)
The Fate of IgE Epitopes and Coeliac Toxic Motifs during Simulated Gastrointestinal Digestion of Pizza Base.
in Foods (Basel, Switzerland)
Geisslitz S
(2021)
Wheat ATIs: Characteristics and Role in Human Disease.
in Frontiers in nutrition
Huang X
(2022)
Quantification of Barley Contaminants in Gluten-Free Oats by Four Gluten ELISA Kits.
in Journal of agricultural and food chemistry
| Description | Overall, the project undertaken during the PhD has developed an in silico approach combined with experimental data on abundance to determine the relative risk of cereals containing gluten in the context of both coeliac disease and IgE mediated allergies. Further work to link this approach with clinical testing data using, for example, challenge meals and incorporating other measures such as T cell binding and IgE cross-reactivity prediction could provide a total measure of the risks associated with different gluten protein types. Sensitivity improvements using enrichment techniques such as antibody enrichment may provide the necessary increase in detection limits to apply the targeted mass spectrometry assay developed in this project for quantitation of gluten in foods. Such an orthogonal test method to ELISA would provide a much needed confirmatory method to support quality assurance of gluten-free foods and ensure, where a product is withdrawn, it is truly warranted. The targets chosen for wheat were also shown to be resistant to processing and simulated digestion, providing another link between analysis and physiological relevance, paving the way for risk-based labelling practices. |
| Exploitation Route | Creation of avenin-free oat lines could be implemented and the ability of these oat lines to elicit symptoms in those with coeliac disease could be evaluated. Further, peptide markers identified in this study could be used for targeted mass spectrometry of gluten in foods however with employment of an enrichment step prior to analysis. |
| Sectors | Agriculture Food and Drink Healthcare |
| Title | Aller-soy |
| Description | Curated soy allergens with evidence of allergenicity present in the WHO/IUIS allergen nomenclature database in FASTA format. |
| Type Of Material | Database/Collection of data |
| Year Produced | 2022 |
| Provided To Others? | Yes |
| URL | https://figshare.com/articles/dataset/Aller-soy/17306822 |
| Title | Aller-wheat |
| Description | Curated wheat allergens with evidence of allergenicity present in the WHO/IUIS allergen nomenclature database in FASTA format. |
| Type Of Material | Database/Collection of data |
| Year Produced | 2022 |
| Provided To Others? | Yes |
| URL | https://figshare.com/articles/dataset/Aller-wheat/17306804 |
| Title | The GluPro suite of curated cereal seed storage prolamin protein sequence datasets |
| Description | Curated protein sequence datasets of the prolamin seed storage proteins present in bread wheat (GluPro v 1.1 and v 1.2), pasta wheat (GluPro v 2 and v 2.1), barley (GluPro v 3), rye (GluPro v 4), oats (GluPro v 5) and a compendium database containing all species sequences (GluPro v 6 and 6.1). These sequences were retrieved from UniProt, NCBI Protein Database, and genomic and transcriptomic data, where available. |
| Type Of Material | Database/Collection of data |
| Year Produced | 2020 |
| Provided To Others? | Yes |
| Impact | The datasets contained within this DOI were cited in a publication "Mapping Coeliac Toxic Motifs in the Prolamin Seed Storage Proteins of Barley, Rye, and Oats Using a Curated Sequence Database, Daly, M., et al., 2020" and will be used for future mining of mass spectrometry data. |
| URL | https://figshare.com/articles/The_GluPro_suite_of_curated_cereal_seed_storage_prolamin_protein_seque... |
| Description | Barley hordein detection in gluten-free oats |
| Organisation | University of Helsinki |
| Country | Finland |
| Sector | Academic/University |
| PI Contribution | Training on and access to our triple quadrupole mass spectrometer, and the design of a targeted proteomic workflow. Academic discussions on how to pick peptide targets and the relevance of gluten-free testing. |
| Collaborator Contribution | Access to oat cultivar seeds and transcriptome data, as well as access to their quadrupole time-of-flight mass spectrometer. Once published, I will be co-author on three publications resulting from this collaboration. |
| Impact | Huang, X., Daly, M., Sontag-Strohm, T., Gethings, L.A. Mills, E.N.C. Barley gluten quantification by marker peptides by QqQ and QToF mass spectrometry. (in preparation) Huang, X., Ahola, H., Daly, M., Nitride, C., Mills, C.M., Sontag-Strohm, T. Determination of residual barley in gluten-free oats by four gluten ELISA kits. Submitted to Journal of Agriculture and Food Chemistry |
| Start Year | 2018 |
| Description | Allergy awareness sessions for Co-op academies |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Schools |
| Results and Impact | Collaborated with the Co-op to create two sessions regarding food allergy in order to increase awareness. The first of which was titled "The impact of food allergy on food allergic people", and "Cleaning after allergens". Each session required a presentation, with narration, to be delivered to the Co-op, that would then be shown within the Co-op academies within the North-West. |
| Year(s) Of Engagement Activity | 2021 |