Person:
Martín De Santos, María Del Rosario

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First Name
María Del Rosario
Last Name
Martín De Santos
URI
https://hdl.handle.net/20.500.14352/80461
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Veterinaria
Department
Nutrición y Ciencia de los Alimentos
Area
Nutrición y Bromatología
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    Development of a new recombinant antibody, selected by phage-display technology from a celiac patient library, for detection of gluten in foods
    (Current Research in Food Science, 2023) García Calvo, Eduardo Rafael; García García, Aina; Rodríguez Gómez, Santiago; Farrais, Sergio; Martín De Santos, María Del Rosario; García Lacarra, Teresa
    Gluten, a group of ethanol-soluble proteins present in the endosperm of cereals, is extensively used in the food industry due to its ability to improve dough properties. However, gluten is also associated with a range of gluten-related diseases (GRDs), such as wheat allergies, celiac disease, and gluten intolerance. The recommended treatment for GRDs patients is a gluten-free diet. To monitor adherence to this diet, it is necessary to develop gluten-detection systems in food products. Among the available methods, immunodetection systems are the most popular due to their simplicity, reproducibility, and accuracy. The aim of this study was to generate novel high-affinity antibodies against gluten to be used as the primary reactant in an enzyme-linked immunosorbent assay (ELISA) test. These antibodies were developed by constructing an immune library from mRNA obtained from two celiac patients with a high humoral response to gluten-related proteins. The resulting library (composed by 1.1x107) was subjected to selection against gliadin using phage display technology. Following several rounds of selection, the Fab-C was selected, and demonstrated good functionality in ELISA tests, presenting a limit of detection of 15 mg/kg for detection of gluten in spiked mixtures and food products. The methodology can discriminate gluten-free products according to the current legislation.
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    Production and Characterization of Novel Fabs Generated from Different Phage Display Libraries as Probes for Immunoassays for Gluten Detection in Food
    (Foods, 2023) García Calvo, Eduardo Rafael; García García, Aina; Rodríguez, Santiago; Takkinen, Kristiina; Martín De Santos, María Del Rosario; García Lacarra, Teresa
    Gluten is the main fraction of wheat proteins. It is widely used in the food industry because of the properties that are generated in the dough, but it is also able to trigger diseases like allergies, autoimmunity processes (such as celiac disease), and intolerances in sensitized persons. The most effective therapy for these diseases is the total avoidance of gluten in the diet because it not only prevents damage but also enhances tissue healing. To ensure the absence of gluten in food products labeled as gluten-free, accurate detection systems, like immunoassays, are required. In this work, four recombinant Fab antibody fragments, selected by phage display technology, were produced and tested for specificity and accuracy against gluten in experimental flour mixtures and commercial food products. A high-affinity probe (Fab-C) was identified and characterized. An indirect ELISA test was developed based on Fab-C that complied with the legal detection limits and could be applied in the assessment of gluten-free diets.