Identificador persistente para citar o vincular este elemento:
http://hdl.handle.net/10553/51919
Campo DC | Valor | idioma |
---|---|---|
dc.contributor.author | Salema, Valencio | en_US |
dc.contributor.author | Marín, Elvira | en_US |
dc.contributor.author | Martínez-Arteaga, Rocio | en_US |
dc.contributor.author | Ruano-Gallego, David | en_US |
dc.contributor.author | Fraile, Sofía | en_US |
dc.contributor.author | Margolles, Yago | en_US |
dc.contributor.author | Teira, Xema | en_US |
dc.contributor.author | Gutiérrez Cabrera, Carlos Javier | en_US |
dc.contributor.author | Bodelón, Gustavo | en_US |
dc.contributor.author | Fernández, Luis Ángel | en_US |
dc.contributor.other | Gutierrez, Carlos | - |
dc.contributor.other | Ruano-Gallego, David | - |
dc.contributor.other | Bodelon, Gustavo | - |
dc.contributor.other | Marin, Elvira | - |
dc.contributor.other | Fernandez Herrero, Luis Angel | - |
dc.contributor.other | SALEMA, VALENCIO | - |
dc.date.accessioned | 2018-11-25T05:25:07Z | - |
dc.date.available | 2018-11-25T05:25:07Z | - |
dc.date.issued | 2013 | en_US |
dc.identifier.issn | 1932-6203 | en_US |
dc.identifier.uri | http://hdl.handle.net/10553/51919 | - |
dc.description.abstract | Screening 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.language | eng | en_US |
dc.publisher | 1932-6203 | - |
dc.relation.ispartof | PLoS ONE | en_US |
dc.source | Plos One[ISSN 1932-6203],v. 8 (9) | en_US |
dc.title | Selection of Single Domain Antibodies from Immune Libraries Displayed on the Surface of E. coli Cells with Two beta-Domains of Opposite Topologies | en_US |
dc.type | info:eu-repo/semantics/Article | en_US |
dc.type | Article | en_US |
dc.identifier.doi | 10.1371/journal.pone.0075126 | en_US |
dc.identifier.scopus | 84884523021 | - |
dc.identifier.isi | 000326520200085 | - |
dcterms.isPartOf | Plos One | - |
dcterms.source | Plos One[ISSN 1932-6203],v. 8 (9) | - |
dc.contributor.authorscopusid | 35096610900 | - |
dc.contributor.authorscopusid | 22035460900 | - |
dc.contributor.authorscopusid | 12778488300 | - |
dc.contributor.authorscopusid | 55786459300 | - |
dc.contributor.authorscopusid | 6508063810 | - |
dc.contributor.authorscopusid | 55862272300 | - |
dc.contributor.authorscopusid | 55861997500 | - |
dc.contributor.authorscopusid | 7202545218 | - |
dc.contributor.authorscopusid | 6506050773 | - |
dc.contributor.authorscopusid | 16238777700 | - |
dc.identifier.issue | e75126 | - |
dc.relation.volume | 8 | en_US |
dc.investigacion | Ingeniería y Arquitectura | en_US |
dc.type2 | Artículo | en_US |
dc.identifier.wos | WOS:000326520200085 | - |
dc.contributor.daisngid | 6332490 | - |
dc.contributor.daisngid | 3474467 | - |
dc.contributor.daisngid | 10018516 | - |
dc.contributor.daisngid | 6207211 | - |
dc.contributor.daisngid | 3004896 | - |
dc.contributor.daisngid | 21014474 | - |
dc.contributor.daisngid | 20092384 | - |
dc.contributor.daisngid | 475608 | - |
dc.contributor.daisngid | 2293170 | - |
dc.contributor.daisngid | 847870 | - |
dc.identifier.investigatorRID | E-2989-2012 | - |
dc.identifier.investigatorRID | P-3734-2018 | - |
dc.identifier.investigatorRID | H-2870-2013 | - |
dc.identifier.investigatorRID | L-1701-2014 | - |
dc.identifier.investigatorRID | D-2713-2009 | - |
dc.identifier.investigatorRID | No ID | - |
dc.utils.revision | Sí | en_US |
dc.date.coverdate | Septiembre 2013 | en_US |
dc.identifier.ulpgc | Sí | en_US |
dc.contributor.buulpgc | BU-ING | en_US |
dc.description.sjr | 1,74 | |
dc.description.jcr | 3,534 | |
dc.description.sjrq | Q1 | |
dc.description.jcrq | Q1 | |
dc.description.scie | SCIE | |
dc.description.erihplus | ERIH PLUS | |
item.grantfulltext | open | - |
item.fulltext | Con texto completo | - |
crisitem.author.dept | GIR IUIBS: Medicina Veterinaria e Investigación Terapéutica | - |
crisitem.author.dept | IU de Investigaciones Biomédicas y Sanitarias | - |
crisitem.author.orcid | 0000-0003-0764-7408 | - |
crisitem.author.parentorg | IU de Investigaciones Biomédicas y Sanitarias | - |
crisitem.author.fullName | Gutiérrez Cabrera,Carlos Javier | - |
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