Generation of an Ovomucoid-Immune scFv Library for the Development of Novel Immunoassays in Hen’s Egg Detection

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dc.contributor.authorRodríguez, Santiago
dc.contributor.authorGarcía García, Aina
dc.contributor.authorGarcía Calvo, Eduardo Rafael
dc.contributor.authorEsteban, Vanesa
dc.contributor.authorPastor Vargas, Carlos
dc.contributor.authorDíaz-Perales, Araceli
dc.contributor.authorGarcía Lacarra, Teresa
dc.contributor.authorMartín, Rosario
dc.date.accessioned2024-01-09T18:37:42Z
dc.date.available2024-01-09T18:37:42Z
dc.date.issued2023
dc.description.abstractHen’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>
dc.description.departmentDepto. de Bioquímica y Biología Molecular
dc.description.facultyFac. de Ciencias Químicas
dc.description.refereedTRUE
dc.description.sponsorshipMinisterio de Ciencia e Innovación (España)
dc.description.sponsorshipComunidad de Madrid
dc.description.statuspub
dc.identifier.citationRodríguez 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.
dc.identifier.doi10.3390/foods12203831
dc.identifier.issn2304-8158
dc.identifier.officialurlhttps://doi.org/10.3390/foods12203831
dc.identifier.urihttps://hdl.handle.net/20.500.14352/92132
dc.issue.number20
dc.journal.titleFoods
dc.language.isoeng
dc.page.initial3831
dc.publisherMPDI
dc.relation.projectIDPID2021-122925OB-I00
dc.relation.projectIDFOODAL S2018/BAA-4574
dc.rightsAttribution-NoDerivatives 4.0 Internationalen
dc.rights.accessRightsopen access
dc.rights.urihttp://creativecommons.org/licenses/by-nd/4.0/
dc.subject.keywordELISA
dc.subject.keywordImmune library
dc.subject.keywordImmune library
dc.subject.keywordEgg detection
dc.subject.ucmCiencias Biomédicas
dc.subject.unesco2412 Inmunología
dc.titleGeneration of an Ovomucoid-Immune scFv Library for the Development of Novel Immunoassays in Hen’s Egg Detection
dc.typejournal article
dc.type.hasVersionVoR
dc.volume.number12
dspace.entity.typePublication
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relation.isAuthorOfPublication.latestForDiscovery901e6544-8ac8-4029-bf26-1d9d096fe78e
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