Enzyme-Linked Immunosorbent Assays for the Detection and Quantitation of Gluten in Cereal-Based Foods

Abstract

Enzyme-linked immunosorbent assays (ELISAs) for gluten quantitation use specific antibodies that interact with epitopes of gluten proteins that trigger an immune response in celiac disease patients. To date, four ELISA methods for gluten quantitation have been validated by international collaborative studies. The Skerritt ELISA method was validated in 2001. Three proprietary gluten ELISA methods have since been developed and were validated between 2010 and 2013: the R5 sandwich ELISA, R5 competitive ELISA, and G12 sandwich ELISA kits. In collaborative studies, the R5 sandwich ELISA had a limit of detection (LOD) of 3 mg of gluten/kg and a relative reproducibility standard deviation (RSDR) of 17 to 27% for samples containing intact gluten, whereas the G12 sandwich ELISA had an LOD of 4 mg of gluten/kg and an RSDR of 19 to 33% in 8 of 9 samples containing intact gluten. The R5 competitive assay targets hydrolyzed gluten and was less sensitive (LOD of 10 mg/kg) and showed higher variation (RSDR of 25 to 37%)...