Generation of an Ovomucoid-Immune scFv Library for the Development of Novel Immunoassays in Hen’s Egg Detection
17 Pág.
| Autores: | , , , , , , , |
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| 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 |
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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 |
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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 Sí |
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info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
| dc.publisher.none.fl_str_mv |
Multidisciplinary Digital Publishing Institute |
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Multidisciplinary Digital Publishing Institute |
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reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC instname:Consejo Superior de Investigaciones Científicas (CSIC) |
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Consejo Superior de Investigaciones Científicas (CSIC) |
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DIGITAL.CSIC. Repositorio Institucional del CSIC |
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DIGITAL.CSIC. Repositorio Institucional del CSIC |
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