Wheat is one of the most important sources of nutrition for humankind. However, the prevalence of inflammatory reactions to wheat, such as celiac disease (CD), wheat allergy and non-celiac gluten sensitivity (NCGS) is increasing and may affect up to 10% of the population. The immunoreactive triggers are gluten proteins in case of CD, both gluten and non-gluten proteins in case of wheat allergy, and most likely α-amylase/trypsin inhibitors (ATIs) in NCGS. The only effective treatment for these entities is a gluten-free diet and improvements of analytical methods are urgently required to protect sensitive individuals.<br/>Therefore, the aim was to advance the detection of immunoreactive cereal proteins using enzyme-linked immunosorbent assays (ELISA) and liquid chromatography mass spectrometry (LC-MS/MS).<br/>First, improved reference materials (RM) for gluten from wheat, rye, barley and oats to be used in gluten analytical method development and validation were developed. Having studied the recognition profiles of the antibodies currently used in gluten ELISA test kits, four novel target sequences for antibody generation were identified and a new kit now enables the comprehensive detection of gluten. Further, targeted LC-MS/MS proteomics methods were developed for ATIs, selected CD-active peptides as well as gluten and the use of well-defined gluten RM allowed the first ever accurate conversion of marker peptide concentrations to gluten contents in different food products. These substantial advances in gluten analytical method development will enhance food safety for CD patients and help elucidate causes for the increasing prevalence of CD, NCGS and allergies in the population.

Advancing the detection of immunoreactive cereal proteins to protect sensitive individuals

2019 Fall National Meeting

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