García Calvo, Eduardo RafaelGarcía García, AinaRodríguez Gómez, SantiagoMartín De Santos, María Del RosarioGarcía Lacarra, Teresa2024-05-242024-05-242024-04-26Garcia-Calvo, E.; García-García, A.; Rodríguez, S.; Martín, R.; García, T. Exploring Gluten Assessment in Marketed Products through a Sandwich ELISA Methodology Based on Novel Recombinant Antibodies. Foods 2024, 13, 1341. https://doi.org/10.3390/ foods130913412304-815810.3390/foods13091341https://hdl.handle.net/20.500.14352/1044212024 Descuento MDPIThis study presents the development of a sandwich ELISA method for gluten detection in foods, using recombinant Fab antibody fragments against gliadin. The Fabs were chemically biotinylated and immobilized on streptavidin-coated plates as capture antibodies, while alkaline phosphatase-conjugated Fabs were used as detection antibodies. Four different gliadin-binding Fabs were tested and the Fab pair Fab8E-4 and Fab-C showed the best compatibility. An indirect sandwich immunoassay, using unmodified Fab8E-4 for capture and Fab-C as the detection antibody, achieved a detection limit of 26 ng/mL of gliadin, corresponding to 10 mg/kg of gluten in foods. No cross-reactivity was observed against 60 gluten-free species commonly used in the food industry. Analysis of 50 commercial products demonstrated consistent results compared to the standard method for gluten detection. The complete lack of cross-reactivity of the developed immunoassay with oat products potentially provides an advantage over other gluten detection systems.engAttribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/journal articlehttps://doi.org/10.3390/foods1309134138731712https://pubmed.ncbi.nlm.nih.gov/38731712/open access613.2.099Food controlGlutenRecombinant FabCeliac diseaseSandwich ELISADietética y nutrición (Medicina)3206 Ciencias de la Nutrición