Identification of high-affinity phage-displayed VH fragments by use of a quartz crystal microbalance with dissipation monitoring

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dc.contributor.authorGómez-Arribas, Lidia
dc.contributor.authorJuste-Dolz, Augusto
dc.contributor.authorPeltomaa, Riikka Johanna
dc.contributor.authorGiménez-Romero, David
dc.contributor.authorMorais, Sergi
dc.contributor.authorBarderas, Rodrigo
dc.contributor.authorCuadrado, Carmen
dc.contributor.authorMaquieira, Ángel
dc.contributor.authorBenito Peña, María Elena
dc.contributor.authorMoreno Bondi, María Cruz
dc.date.accessioned2024-01-16T15:11:16Z
dc.date.available2024-01-16T15:11:16Z
dc.date.issued2021
dc.description.abstractPhage display has become a powerful tool for antibody discovery in a wide variety of fields. This technology allows specific binders for a given antigen to be selected from combinatorial libraries. A key step in the process is characterizing and evaluating antibody clones thus selected to reliably identify the best antigen binders. Novel characterization methods can provide essential insight into the binding mechanism and supplement the information obtained with conventional techniques. In this work, we used a quartz crystal microbalance with dissipation monitoring (QCM-D) to determine the kinetic and thermodynamic binding parameters for phage-displayed VH antibody fragments. Phytohemagglutinin (PHA), a legume lectin of analytical interest, was used as a complex model antigen to select specific VH fragments from a phage-displayed library. Eight VH fragments with a unique amino acid sequence were identified as PHA binders by using the well-established enzyme-linked immunosorbent assay (ELISA). QCM-D measurements, structural analysis and principal component analysis (PCA) were used to evaluate the antibody fragments and identify clone clusters with similar binding characteristics and molecular interaction mechanisms. This unprecedented study has enabled the identification of high-affinity phage-displayed VH antibody fragments for PHA, which could be useful for PHA analysis (apparent association constant ranged from 10e8 to 10e10 1/M). In fact, the proposed methodology provides a useful tool for evaluating and characterizing antibody fragments with capabilities beyond those of conventional techniques.
dc.description.departmentDepto. de Química Analítica
dc.description.facultyFac. de Ciencias Químicas
dc.description.refereedTRUE
dc.description.sponsorshipEuropean Commission
dc.description.statuspub
dc.identifier.citationGómez-Arribas, L. N.; Juste-Dolz, A.; Peltomaa, R.; Giménez-Romero, D.; Morais, S.; Barderas, R.; Cuadrado, C.; Maquieira, Á; Benito-Peña, E.; Moreno-Bondi, M. C. Identification of high-affinity phage-displayed VH fragments by use of a quartz crystal microbalance with dissipation monitoring. Sensors Actuators B: Chem. 2021, 340, 129954 DOI:10.1016/j.snb.2021.129954.
dc.identifier.doi10.1016/j.snb.2021.129954
dc.identifier.issn0925-4005
dc.identifier.officialurlhttps://doi.org/10.1016/j.snb.2021.129954
dc.identifier.relatedurlhttps://www.sciencedirect.com/science/article/pii/S0925400521005232?via%3Dihub
dc.identifier.urihttps://hdl.handle.net/20.500.14352/93413
dc.issue.number129954
dc.journal.titleSensors and Actuators: B. Chemical
dc.language.isoeng
dc.page.final10
dc.page.initial1
dc.publisherElsevier
dc.relation.projectIDinfo:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/RTI2018-096410-B-C21/ES/MATERIALES BIO(MIMETICOS) INNOVADORES PARA SENSORES OPTICOS Y SEPARACIONES ANALITICAS/
dc.relation.projectIDCTQ2016-75749-R
dc.relation.projectIDGVA’s Prometeo program (Grant 2020/094)
dc.rightsAttribution-NonCommercial-NoDerivatives 4.0 Internationalen
dc.rights.accessRightsopen access
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subject.cdu547
dc.subject.keywordPhage display
dc.subject.keywordAntibody fragment
dc.subject.keywordQuartz crystal microbalance
dc.subject.keywordBiosensor
dc.subject.keywordPrincipal component analysis
dc.subject.keywordLectins
dc.subject.ucmCiencias
dc.subject.unesco23 Química
dc.subject.unesco33 Ciencias Tecnológicas
dc.subject.unesco32 Ciencias Médicas
dc.subject.unesco31 Ciencias Agrarias
dc.titleIdentification of high-affinity phage-displayed VH fragments by use of a quartz crystal microbalance with dissipation monitoring
dc.typejournal article
dc.type.hasVersionSMUR
dc.volume.number340
dspace.entity.typePublication
relation.isAuthorOfPublication4f6d77c5-1abc-4cab-9a5c-eacb835fce30
relation.isAuthorOfPublicationebb3e2fd-e5d5-4a84-9ce0-c9ea12eb2a85
relation.isAuthorOfPublication8766057b-6628-4a02-a6db-20bddfaf3054
relation.isAuthorOfPublication.latestForDiscovery8766057b-6628-4a02-a6db-20bddfaf3054
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