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

17 Pág.

Detalles Bibliográficos
Autores: Rodríguez, Santiago, García-García, Aina, García-Calvo, Eduardo, Esteban, Vanesa, Pastor-Vargas, C., Díaz-Perales, Araceli, García, Teresa, Martín, Rosario
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2023
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/349026
Acceso en línea:http://hdl.handle.net/10261/349026
https://api.elsevier.com/content/abstract/scopus_id/85175195624
Access Level:acceso abierto
Palabra clave:Egg detection
ELISA
Immune library
Ovomucoid
Phage display
Recombinant antibody
scFv
id ES_aa796d4d73153fd4e4ebd1be76db7907
oai_identifier_str oai:digital.csic.es:10261/349026
network_acronym_str ES
network_name_str España
repository_id_str
spelling Generation of an Ovomucoid-Immune scFv Library for the Development of Novel Immunoassays in Hen’s Egg DetectionRodríguez, SantiagoGarcía-García, AinaGarcía-Calvo, EduardoEsteban, VanesaPastor-Vargas, C.Díaz-Perales, AraceliGarcía, TeresaMartín, RosarioEgg detectionELISAImmune libraryOvomucoidPhage displayRecombinant antibodyscFv17 Pág.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.This research was funded by Ministerio de Ciencia e Innovación Grant: PID2021-122925OB-I00 and Consejería de Educación e Investigación de la Comunidad de Madrid [FOODAL S2018/BAA-4574].Peer reviewedMultidisciplinary Digital Publishing InstituteMinisterio de Ciencia e Innovación (España)Agencia Estatal de Investigación (España)Comunidad de MadridRodríguez, Santiago [0000-0003-2185-1528]García-García, Aina [0000-0003-1689-8751]García-Calvo, E. [0000-0001-8260-9976]Esteban, Vanesa [0000-0002-7500-6277]Pastor-Vargas, C. [0000-0002-4678-7967]Díaz-Perales, Araceli [0000-0002-1093-3627]García, Teresa [0000-0001-7387-0603]Martín, RosarioConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202420242023info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/349026https://api.elsevier.com/content/abstract/scopus_id/85175195624reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023/PID2021-122925OB-I00S2018/BAA-4574/FOODAL-CMCentro de Biotecnología y Genómica de Plantas (CBGP)https://doi.org/10.3390/foods12203831Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/3490262026-05-22T06:33:51Z
dc.title.none.fl_str_mv Generation of an Ovomucoid-Immune scFv Library for the Development of Novel Immunoassays in Hen’s Egg Detection
title Generation of an Ovomucoid-Immune scFv Library for the Development of Novel Immunoassays in Hen’s Egg Detection
spellingShingle Generation of an Ovomucoid-Immune scFv Library for the Development of Novel Immunoassays in Hen’s Egg Detection
Rodríguez, Santiago
Egg detection
ELISA
Immune library
Ovomucoid
Phage display
Recombinant antibody
scFv
title_short Generation of an Ovomucoid-Immune scFv Library for the Development of Novel Immunoassays in Hen’s Egg Detection
title_full Generation of an Ovomucoid-Immune scFv Library for the Development of Novel Immunoassays in Hen’s Egg Detection
title_fullStr Generation of an Ovomucoid-Immune scFv Library for the Development of Novel Immunoassays in Hen’s Egg Detection
title_full_unstemmed Generation of an Ovomucoid-Immune scFv Library for the Development of Novel Immunoassays in Hen’s Egg Detection
title_sort Generation of an Ovomucoid-Immune scFv Library for the Development of Novel Immunoassays in Hen’s Egg Detection
dc.creator.none.fl_str_mv Rodríguez, Santiago
García-García, Aina
García-Calvo, Eduardo
Esteban, Vanesa
Pastor-Vargas, C.
Díaz-Perales, Araceli
García, Teresa
Martín, Rosario
author Rodríguez, Santiago
author_facet Rodríguez, Santiago
García-García, Aina
García-Calvo, Eduardo
Esteban, Vanesa
Pastor-Vargas, C.
Díaz-Perales, Araceli
García, Teresa
Martín, Rosario
author_role author
author2 García-García, Aina
García-Calvo, Eduardo
Esteban, Vanesa
Pastor-Vargas, C.
Díaz-Perales, Araceli
García, Teresa
Martín, Rosario
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Ministerio de Ciencia e Innovación (España)
Agencia Estatal de Investigación (España)
Comunidad de Madrid
Rodríguez, Santiago [0000-0003-2185-1528]
García-García, Aina [0000-0003-1689-8751]
García-Calvo, E. [0000-0001-8260-9976]
Esteban, Vanesa [0000-0002-7500-6277]
Pastor-Vargas, C. [0000-0002-4678-7967]
Díaz-Perales, Araceli [0000-0002-1093-3627]
García, Teresa [0000-0001-7387-0603]
Martín, Rosario
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Egg detection
ELISA
Immune library
Ovomucoid
Phage display
Recombinant antibody
scFv
topic Egg detection
ELISA
Immune library
Ovomucoid
Phage display
Recombinant antibody
scFv
description 17 Pág.
publishDate 2023
dc.date.none.fl_str_mv 2023
2024
2024
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/349026
https://api.elsevier.com/content/abstract/scopus_id/85175195624
url http://hdl.handle.net/10261/349026
https://api.elsevier.com/content/abstract/scopus_id/85175195624
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023/PID2021-122925OB-I00
S2018/BAA-4574/FOODAL-CM
Centro de Biotecnología y Genómica de Plantas (CBGP)
https://doi.org/10.3390/foods12203831

dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869416184719343616
score 15,81155