Celiac disease (CD) affects approximately 1 in 141 individuals in the United States, requiring adherence to a strict gluten-free diet. The Codex Standard, the European Commission, and the FDA all state that any foods that carry the label “gluten-free” must contain less than 20 parts per million (ppm) of gluten. Accurate quantitation of gluten in fermented-hydrolyzed foods is challenging due to the lack of appropriate reference materials and variable proteolysis. A novel multiplex-competitive ELISA method was recently developed utilizing the G12, R5, 2D4, MIoBS, and Skerritt antibody-HRP conjugates employed in nine commercial gluten ELISAs. The multiplex assay is able to reliably distinguish between different forms of fermentation / hydrolysis; thereby serving a unique role necessary for the selection of appropriate calibration standards and, ultimately, the accurate quantitation of hydrolyzed gluten. To further assess the potential utility of this unique multiplex-competitive ELISA, its performance with yogurt prepared with gluten incurred at four concentrations (0, 20, 100, and 500 µg/g) was examined. Gluten incurred in yogurt is partially proteolyzed during the fermentation process, providing a suitable matrix to assess the ability of the multiplex-competitive ELISA to differentiate foods based on the degrees of fermentation, in addition to its ability to differentiate foods based on the types/forms of fermentation. This is essential for the selection of an appropriate calibrator, as not all fermented-hydrolyzed foods are proteolyzed to the same degree during the production process, which leads to differences in protein/peptide profiles. Results indicated a unique antibody-dependent response when gluten incurred in yogurt was fermented, as well as similar gluten proteolysis among the yogurts at different gluten concentrations. To evaluate the reliability and reproducibility of the multiplex-competitive ELISA, a multi-lab/analyst evaluation using the prepared yogurt as a model food is currently underway.<br/>

Immunochemical methods for detecting fermented-hydrolyzed gluten in food: Challenges and recent progress

ACS Fall 2020 Virtual Meeting

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