Identificador persistente para citar o vincular este elemento: http://hdl.handle.net/10553/51919
Campo DC Valoridioma
dc.contributor.authorSalema, Valencioen_US
dc.contributor.authorMarín, Elviraen_US
dc.contributor.authorMartínez-Arteaga, Rocioen_US
dc.contributor.authorRuano-Gallego, Daviden_US
dc.contributor.authorFraile, Sofíaen_US
dc.contributor.authorMargolles, Yagoen_US
dc.contributor.authorTeira, Xemaen_US
dc.contributor.authorGutiérrez Cabrera, Carlos Javieren_US
dc.contributor.authorBodelón, Gustavoen_US
dc.contributor.authorFernández, Luis Ángelen_US
dc.contributor.otherGutierrez, Carlos-
dc.contributor.otherRuano-Gallego, David-
dc.contributor.otherBodelon, Gustavo-
dc.contributor.otherMarin, Elvira-
dc.contributor.otherFernandez Herrero, Luis Angel-
dc.contributor.otherSALEMA, VALENCIO-
dc.date.accessioned2018-11-25T05:25:07Z-
dc.date.available2018-11-25T05:25:07Z-
dc.date.issued2013en_US
dc.identifier.issn1932-6203en_US
dc.identifier.urihttp://hdl.handle.net/10553/51919-
dc.description.abstractScreening of antibody (Ab) libraries by direct display on the surface of E. coli cells is hampered by the presence of the outer membrane (OM). In this work we demonstrate that the native β-domains of EhaA autotransporter and intimin, two proteins from enterohemorrhagic E. coli O157:H7 (EHEC) with opposite topologies in the OM, are effective systems for the display of immune libraries of single domain Abs (sdAbs) from camelids (nanobodies or VHH) on the surface of E. coli K-12 cells and for the selection of high affinity sdAbs using magnetic cell sorting (MACS). We analyzed the capacity of EhaA and intimin β-domains to display individual sdAbs and sdAb libraries obtained after immunization with the extracellular domain of the translocated intimin receptor from EHEC (TirMEHEC). We demonstrated that both systems displayed functional sdAbs on the surface of E. coli cells with little proteolysis and cellular toxicity, although E. coli cells displaying sdAbs with the β-domain of intimin showed higher antigen-binding capacity. Both E. coli display libraries were screened for TirMEHEC binding clones by MACS. High affinity binders were selected by both display systems, although more efficiently with the intimin β-domain. The specificity of the selected clones against TirMEHEC was demonstrated by flow cytometry of E. coli cells, along with ELISA and surface plasmon resonance with purified sdAbs. Finally, we employed the E. coli cell display systems to provide an estimation of the affinity of the selected sdAb by flow cytometry analysis under equilibrium conditions. © 2013 Salema et al.en_US
dc.languageengen_US
dc.publisher1932-6203-
dc.relation.ispartofPLoS ONEen_US
dc.sourcePlos One[ISSN 1932-6203],v. 8 (9)en_US
dc.titleSelection of Single Domain Antibodies from Immune Libraries Displayed on the Surface of E. coli Cells with Two beta-Domains of Opposite Topologiesen_US
dc.typeinfo:eu-repo/semantics/Articleen_US
dc.typeArticleen_US
dc.identifier.doi10.1371/journal.pone.0075126en_US
dc.identifier.scopus84884523021-
dc.identifier.isi000326520200085-
dcterms.isPartOfPlos One-
dcterms.sourcePlos One[ISSN 1932-6203],v. 8 (9)-
dc.contributor.authorscopusid35096610900-
dc.contributor.authorscopusid22035460900-
dc.contributor.authorscopusid12778488300-
dc.contributor.authorscopusid55786459300-
dc.contributor.authorscopusid6508063810-
dc.contributor.authorscopusid55862272300-
dc.contributor.authorscopusid55861997500-
dc.contributor.authorscopusid7202545218-
dc.contributor.authorscopusid6506050773-
dc.contributor.authorscopusid16238777700-
dc.identifier.issuee75126-
dc.relation.volume8en_US
dc.investigacionIngeniería y Arquitecturaen_US
dc.type2Artículoen_US
dc.identifier.wosWOS:000326520200085-
dc.contributor.daisngid6332490-
dc.contributor.daisngid3474467-
dc.contributor.daisngid10018516-
dc.contributor.daisngid6207211-
dc.contributor.daisngid3004896-
dc.contributor.daisngid21014474-
dc.contributor.daisngid20092384-
dc.contributor.daisngid475608-
dc.contributor.daisngid2293170-
dc.contributor.daisngid847870-
dc.identifier.investigatorRIDE-2989-2012-
dc.identifier.investigatorRIDP-3734-2018-
dc.identifier.investigatorRIDH-2870-2013-
dc.identifier.investigatorRIDL-1701-2014-
dc.identifier.investigatorRIDD-2713-2009-
dc.identifier.investigatorRIDNo ID-
dc.utils.revisionen_US
dc.date.coverdateSeptiembre 2013en_US
dc.identifier.ulpgcen_US
dc.contributor.buulpgcBU-INGen_US
dc.description.sjr1,74
dc.description.jcr3,534
dc.description.sjrqQ1
dc.description.jcrqQ1
dc.description.scieSCIE
dc.description.erihplusERIH PLUS
item.grantfulltextopen-
item.fulltextCon texto completo-
crisitem.author.deptGIR IUIBS: Medicina Veterinaria e Investigación Terapéutica-
crisitem.author.deptIU de Investigaciones Biomédicas y Sanitarias-
crisitem.author.orcid0000-0003-0764-7408-
crisitem.author.parentorgIU de Investigaciones Biomédicas y Sanitarias-
crisitem.author.fullNameGutiérrez Cabrera,Carlos Javier-
Colección:Artículos
miniatura
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