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

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dc.contributor.authorGarcía Calvo, Eduardo Rafael
dc.contributor.authorGarcía García, Aina
dc.contributor.authorRodríguez Gómez, Santiago
dc.contributor.authorMartín De Santos, María Del Rosario
dc.contributor.authorGarcía Lacarra, Teresa
dc.date.accessioned2024-06-10T14:09:21Z
dc.date.available2024-06-10T14:09:21Z
dc.date.issued2024-04-26
dc.description.abstractDirected 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.
dc.description.departmentDepto. de Nutrición y Ciencia de los Alimentos
dc.description.facultyFac. de Veterinaria
dc.description.refereedTRUE
dc.description.sponsorshipMinisterio de Ciencia e Innovación (España)
dc.description.statuspub
dc.identifier.citationGarcia-Calvo, E.; García-García, A.; Rodríguez, S.; Martín, R.; García, T. Unraveling the Properties of Phage Display Fab Libraries and Their Use in the Selection of Gliadin-Specific Probes by Applying High-Throughput Nanopore Sequencing. Viruses 2024, 16, 686. https://doi.org/10.3390/ v16050686
dc.identifier.doi10.3390/v16050686
dc.identifier.issn1999-4915
dc.identifier.officialurlhttps://doi.org/10.3390/v16050686
dc.identifier.pmid38793567
dc.identifier.relatedurlhttps://www.mdpi.com/journal/viruses
dc.identifier.urihttps://hdl.handle.net/20.500.14352/104803
dc.issue.number686
dc.journal.titleViruses
dc.language.isoeng
dc.page.final18
dc.page.initial1
dc.publisherMDPI
dc.relation.projectIDinfo:eu-repo/grantAgreement/AEI/GENERACIÓN DE REPERTORIOS DE ANTICUERPOS RECOMBINANTES PARA EL DESARROLLO DE INMUNOENSAYOS DESTINADOS AL CONTROL DE ALÉRGENOS EN LA INDUSTRIA ALIMENTARIAPID2021-122925OB-I00
dc.rightsAttribution 4.0 Internationalen
dc.rights.accessRightsopen access
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subject.cdu641
dc.subject.keywordCeliac disease
dc.subject.keywordPhage display
dc.subject.keywordHigh-throughput DNA sequencing
dc.subject.keywordNanopore sequencing
dc.subject.keywordGliadin
dc.subject.ucmAlimentación
dc.subject.unesco3206 Ciencias de la Nutrición
dc.titleUnraveling the Properties of Phage Display Fab Libraries and Their Use in the Selection of Gliadin-Specific Probes by Applying High-Throughput Nanopore Sequencing
dc.typejournal article
dc.type.hasVersionVoR
dc.volume.number16
dspace.entity.typePublication
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Unraveling the Properties of Phage Display Fab Libraries and Their Use in the Selection of Gliadin-Specific Probes by Applying High-Throughput Nanopore Sequencing
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