The overall objective of this project is the development of a disposable microsystem with integrated optimal extraction and detection for the precise and accurate quantification of coeliac disease toxic gluten (sequences of gluten to which coeliac disease sufferers show intolerance) in all types of foodstuffs. Screening studies currently being carried out both in Europe (CD-Cluster project) as well as in the United States are highlighting that the occurrence of coeliac disease is far higher than previously believed. This will inevitably lead to an increased control and the allowable limits and assay for measurement recommended by the WHO/FAO Codex Alimentarius Commission will be internationally recognised and implemented and it is thus crucial, in terms of European competitiveness, that this be a European based product.
The work proposed here not only addresses the problems previously observed with gluten assays, but also aims at the simplifying of the assay to a biosensoric format: facile, inexpensive and rapid, which will be integrated with a microextraction module in a disposable microsystem allowing complete assay within 15 minutes on site or on line. It is envisaged that the proposed technology can be used to monitor quality throughout the processing of gluten-free ingredients and production of gluten free foods, for use by regulatory authorities (e.g. the proposed European Food Authority), by vendors of gluten free food, and eventually, with high production resulting in lowered costs, by the coeliac disease sufferer themselves.
- The development of detection-compatible protocols for extraction of gluten from all types of foodstuffs.
- The production of aptamers, antibody cocktails and antibody fragments to toxic sequences identified by in vitro and in vivo testing
- The isolation and purification of solvent resistant thermophilic enzymes for use as aptamer/antibody labels
- The development of ELISAs/ ELONAs /mixed ELISA-ELONAs.
- The development of immuno- and apta- sensor generic platforms with optical/electrochemical/quartz crystal nanobalance detection
- The integration of extraction and detection on a disposable microsystem with the time required for total assay being less than 15 minutes and the cost less than _15, meeting the PDRs listed both for extraction and for detection.
- The analytical validation, end-user evaluation and dissemination of the developed technologies.
- Standardised and fully validated protocols and universal protocol for the efficient extraction of gluten from raw, cooked, unprocessed and processed foodstuffs
- Standardised and fully validated assay based on aptamers and antibodies/fragments for the selective, specific, and sensitive determination of CD-toxic gluten from all cultivars of wheat, barley, rye and oats with no cross-reactivity
- Standardised and fully validated assay based on aptamers and antibodies/fragments compatible with extraction solvents facilitating detection of CD-toxic gluten in raw, cooked, unprocessed and processed foodstuffs
- Standardised and fully validated immuno-/apta- sensor platforms for the detection of CD-toxic gluten in raw, cooked, unprocessed and processed foodstuffs using electrochemical, optical and quartz crystal nanobalance transduction
- Prototype biosensor immuno-/apta- sensor platform integratable with microsystem, based on optimum mode of transduction
- Fully validated microsystem extraction of CD-toxic gluten from raw, cooked, unprocessed and processed foodstuffs
- Fully validated disposable microsystem with integrated extraction and biosensoric detection
- End user evaluation and feedback of project products
- Consumer-science studies - feedback on increased consumer confidence based on project's developed technologies
- Technology Implementation Plan and widespread dissemination