Unraveling the Properties of Phage Display Fab Libraries and Their Use in the Selection of Gliadin-Specific Probes by Applying High-Throughput Nanopore Sequencing

Directed evolution is a pivotal strategy for new antibody discovery, which allowed the generation of high-affinity Fabs against gliadin from two antibody libraries in our previous studies. One of the libraries was exclusively derived from celiac patients’ mRNA (immune library) while the other was ob...

Descripción completa

Detalles Bibliográficos
Autores: García Calvo, Eduardo Rafael, García García, Aina, Rodríguez Gómez, Santiago, Martín De Santos, María Del Rosario, García Lacarra, Teresa
Tipo de recurso: artículo
Fecha de publicación:2024
País:España
Institución:Universidad Complutense de Madrid (UCM)
Repositorio:Docta Complutense
Idioma:inglés
OAI Identifier:oai:docta.ucm.es:20.500.14352/104803
Acceso en línea:https://hdl.handle.net/20.500.14352/104803
Access Level:acceso abierto
Palabra clave:641
Celiac disease
Phage display
High-throughput DNA sequencing
Nanopore sequencing
Gliadin
Alimentación
3206 Ciencias de la Nutrición
id ES_16b21c8677bf4e93ef8ecd594f5bc9d9
oai_identifier_str oai:docta.ucm.es:20.500.14352/104803
network_acronym_str ES
network_name_str España
repository_id_str
spelling Unraveling the Properties of Phage Display Fab Libraries and Their Use in the Selection of Gliadin-Specific Probes by Applying High-Throughput Nanopore SequencingGarcía Calvo, Eduardo RafaelGarcía García, AinaRodríguez Gómez, SantiagoMartín De Santos, María Del RosarioGarcía Lacarra, Teresa641Celiac diseasePhage displayHigh-throughput DNA sequencingNanopore sequencingGliadinAlimentación3206 Ciencias de la NutriciónDirected evolution is a pivotal strategy for new antibody discovery, which allowed the generation of high-affinity Fabs against gliadin from two antibody libraries in our previous studies. One of the libraries was exclusively derived from celiac patients’ mRNA (immune library) while the other was obtained through a protein engineering approach (semi-immune library). Recent advances in high-throughput DNA sequencing techniques are revolutionizing research across genomics, epigenomics, and transcriptomics. In the present work, an Oxford Nanopore in-lab sequencing device was used to comprehensively characterize the composition of the constructed libraries, both at the beginning and throughout the phage-mediated selection processes against gliadin. A customized analysis pipeline was used to select high-quality reads, annotate chain distribution, perform sequence analysis, and conduct statistical comparisons between the different selection rounds. Some immunological attributes of the most representative phage variants after the selection process were also determined. Sequencing results revealed the successful transfer of the celiac immune response features to the immune library and the antibodies derived from it, suggesting the crucial role of these features in guiding the selection of high-affinity recombinant Fabs against gliadin. In summary, high-throughput DNA sequencing has improved our understanding of the selection processes aimed at generating molecular binders against gliadin.MDPIUniversidad Complutense de Madrid20242024-04-2620242024-04-26journal articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/20.500.14352/104803reponame:Docta Complutenseinstname:Universidad Complutense de Madrid (UCM)InglésengAgencia Estatal de Investigación http://dx.doi.org/10.13039/501100011033 GENERACIÓN DE REPERTORIOS DE ANTICUERPOS RECOMBINANTES PARA EL DESARROLLO DE INMUNOENSAYOS DESTINADOS AL CONTROL DE ALÉRGENOS EN LA INDUSTRIA ALIMENTARIAPID2021-122925OB-I00 Not availableopen accesshttp://purl.org/coar/access_right/c_abf2Attribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:docta.ucm.es:20.500.14352/1048032026-06-02T12:44:21Z
dc.title.none.fl_str_mv Unraveling the Properties of Phage Display Fab Libraries and Their Use in the Selection of Gliadin-Specific Probes by Applying High-Throughput Nanopore Sequencing
title Unraveling the Properties of Phage Display Fab Libraries and Their Use in the Selection of Gliadin-Specific Probes by Applying High-Throughput Nanopore Sequencing
spellingShingle Unraveling the Properties of Phage Display Fab Libraries and Their Use in the Selection of Gliadin-Specific Probes by Applying High-Throughput Nanopore Sequencing
García Calvo, Eduardo Rafael
641
Celiac disease
Phage display
High-throughput DNA sequencing
Nanopore sequencing
Gliadin
Alimentación
3206 Ciencias de la Nutrición
title_short Unraveling the Properties of Phage Display Fab Libraries and Their Use in the Selection of Gliadin-Specific Probes by Applying High-Throughput Nanopore Sequencing
title_full Unraveling the Properties of Phage Display Fab Libraries and Their Use in the Selection of Gliadin-Specific Probes by Applying High-Throughput Nanopore Sequencing
title_fullStr Unraveling the Properties of Phage Display Fab Libraries and Their Use in the Selection of Gliadin-Specific Probes by Applying High-Throughput Nanopore Sequencing
title_full_unstemmed Unraveling the Properties of Phage Display Fab Libraries and Their Use in the Selection of Gliadin-Specific Probes by Applying High-Throughput Nanopore Sequencing
title_sort Unraveling the Properties of Phage Display Fab Libraries and Their Use in the Selection of Gliadin-Specific Probes by Applying High-Throughput Nanopore Sequencing
dc.creator.none.fl_str_mv García Calvo, Eduardo Rafael
García García, Aina
Rodríguez Gómez, Santiago
Martín De Santos, María Del Rosario
García Lacarra, Teresa
author García Calvo, Eduardo Rafael
author_facet García Calvo, Eduardo Rafael
García García, Aina
Rodríguez Gómez, Santiago
Martín De Santos, María Del Rosario
García Lacarra, Teresa
author_role author
author2 García García, Aina
Rodríguez Gómez, Santiago
Martín De Santos, María Del Rosario
García Lacarra, Teresa
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Universidad Complutense de Madrid
dc.subject.none.fl_str_mv 641
Celiac disease
Phage display
High-throughput DNA sequencing
Nanopore sequencing
Gliadin
Alimentación
3206 Ciencias de la Nutrición
topic 641
Celiac disease
Phage display
High-throughput DNA sequencing
Nanopore sequencing
Gliadin
Alimentación
3206 Ciencias de la Nutrición
description Directed evolution is a pivotal strategy for new antibody discovery, which allowed the generation of high-affinity Fabs against gliadin from two antibody libraries in our previous studies. One of the libraries was exclusively derived from celiac patients’ mRNA (immune library) while the other was obtained through a protein engineering approach (semi-immune library). Recent advances in high-throughput DNA sequencing techniques are revolutionizing research across genomics, epigenomics, and transcriptomics. In the present work, an Oxford Nanopore in-lab sequencing device was used to comprehensively characterize the composition of the constructed libraries, both at the beginning and throughout the phage-mediated selection processes against gliadin. A customized analysis pipeline was used to select high-quality reads, annotate chain distribution, perform sequence analysis, and conduct statistical comparisons between the different selection rounds. Some immunological attributes of the most representative phage variants after the selection process were also determined. Sequencing results revealed the successful transfer of the celiac immune response features to the immune library and the antibodies derived from it, suggesting the crucial role of these features in guiding the selection of high-affinity recombinant Fabs against gliadin. In summary, high-throughput DNA sequencing has improved our understanding of the selection processes aimed at generating molecular binders against gliadin.
publishDate 2024
dc.date.none.fl_str_mv 2024
2024-04-26
2024
2024-04-26
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://hdl.handle.net/20.500.14352/104803
url https://hdl.handle.net/20.500.14352/104803
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.relation.none.fl_str_mv Agencia Estatal de Investigación http://dx.doi.org/10.13039/501100011033 GENERACIÓN DE REPERTORIOS DE ANTICUERPOS RECOMBINANTES PARA EL DESARROLLO DE INMUNOENSAYOS DESTINADOS AL CONTROL DE ALÉRGENOS EN LA INDUSTRIA ALIMENTARIAPID2021-122925OB-I00 Not available
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv reponame:Docta Complutense
instname:Universidad Complutense de Madrid (UCM)
instname_str Universidad Complutense de Madrid (UCM)
reponame_str Docta Complutense
collection Docta Complutense
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869403870474534912
score 15,300719

Similares en LA Referencia