Purpose: 1) To validate the performance of the MonoTraceTMGluten ELISA using both raw and processed foods. 2) To test this monoclonal antibody-based assay for cross-reactivity (CR) against a large panel of cereals, tree nuts, legumes, seeds, meats and spices.
Methods: The assay limit of detection (LOD) was calculated by testing kit standards in triplicate on three different days. Recovery from various food matrices and ingredients (n=18) was analyzed using in-house incurred cornbread samples, gluten-free (GF) commercial products and ingredients, as wells as quality control and reference materials sourced from the Food Analysis Performance Assessment Scheme (FAPAS) division of the Food and Environment Research Agency (FERA). Cross-reactivity was tested using a large panel of GF and nonGF labeled commercial samples from various vendors (n=126).
Results: The LOD for this new commercial gluten ELISA was determined statistically to be 0.3 ppm and the range of quantification (ROQ) was determined to be 2 to 100 ppm gluten (1 to 50 ppm gliadin). Recovery of gluten from in-house gluten-incurred cornbread and gluten-spiked commercial food matrices was >75%. No cross-reactivity was observed in any of the tree nuts, legumes, seeds, meats or spice samples tested. Wheat, rye and barley flours were highly CR. The assay also exhibits little to no cross-reactivity to several oat cultivars (n=5).
Significance: The BioFront Technologies’ gluten ELISA represents a rapid method for detecting and accurately quantifying gluten contamination within raw food ingredients and processed foods. The assay is highly specific, sensitive and has a wide ROQ.