RT Journal Article
T1 Generation of an Ovomucoid-Immune scFv Library for the Development of Novel Immunoassays in Hen’s Egg Detection
A1 Rodríguez, Santiago
A1 García García, Aina
A1 García Calvo, Eduardo Rafael
A1 Esteban, Vanesa
A1 Pastor Vargas, Carlos
A1 Díaz-Perales, Araceli
A1 García Lacarra, Teresa
A1 Martín, Rosario
AB Hen’s egg allergy is the second most common food allergy among infants and young children. The possible presence of undeclared eggs in foods poses a significant risk to sensitized individuals. Therefore, reliable egg allergen detection methods are needed to ensure compliance with food labeling and improve consumer protection. This work describes for the first time the application of phage display technology for the generation of a recombinant antibody aimed at the specific detection of hen’s ovomucoid. First, a single-chain variable fragment (scFv) library was constructed from mRNA isolated from the spleen of a rabbit immunized with ovomucoid. After rounds of biopanning, four binding clones were isolated and characterized. Based on the best ovomucoid-binding candidate SR-G1, an indirect phage enzyme-linked immunosorbent assay (phage-ELISA) was developed, reaching limits of detection and quantitation of 43 and 79 ng/mL of ovomucoid, respectively. The developed ELISA was applied to the analysis of a wide variety of food products, obtaining a good correlation with a commercial egg detection assay used as a reference. Finally, in silico modeling of the antigen-antibody complex revealed that the main interactions most likely occur between the scFv heavy chain and the ovomucoid domain-III, the most immunogenic region of this allergen.</jats:p>
PB MPDI
SN 2304-8158
YR 2023
FD 2023
LK https://hdl.handle.net/20.500.14352/92132
UL https://hdl.handle.net/20.500.14352/92132
LA eng
NO Rodríguez S, García-García A, Garcia-Calvo E, Esteban V, Pastor-Vargas C, Díaz-Perales A, García T, Martín R. Generation of an Ovomucoid-Immune scFv Library for the Development of Novel Immunoassays in Hen's Egg Detection. Foods. 2023 Oct 19;12(20):3831. doi: 10.3390/foods12203831. PMID: 37893724; PMCID: PMC10606182.
NO Ministerio de Ciencia e Innovación (España)
NO Comunidad de Madrid
DS Docta Complutense
RD 7 abr 2025