{"id":585,"date":"2016-09-20T18:53:06","date_gmt":"2016-09-20T18:53:06","guid":{"rendered":"http:\/\/medicinaveterinaria.ulpgc.es\/?p=585"},"modified":"2016-10-17T15:44:46","modified_gmt":"2016-10-17T15:44:46","slug":"characterization-of-nanobodies-binding-human-fibrinogen-selected-by-e-coli-display","status":"publish","type":"post","link":"https:\/\/medicinaveterinaria.ulpgc.es\/?p=585","title":{"rendered":"Characterization of nanobodies binding human fibrinogen selected by E. coli display\u00a0"},"content":{"rendered":"<div class=\"paddingT5 paddingB5\">\n<div class=\"sourceCrossMain\">\n<div class=\"sourceTitle sourceTitleWidth\">Journal of Biotechnology<\/div>\n<\/div>\n<div class=\"volumeInfo\">Volume 234, 20 September 2016, Pages 58-65<\/div>\n<div><\/div>\n<\/div>\n<div class=\"hr\">\u00a0Salema, V.a,\u00a0L\u00f3pez-Guajardo, A.a, \u00a0Gutierrez, C.b,\u00a0Menc\u00eda, M.c, \u00a0Fern\u00e1ndez, L.\u00c1.a<\/div>\n<div>\n<div id=\"authorlist\" class=\"smallLink authorlink svDoNotLink displayNone\">\n<div class=\"clear floatNone\"><span id=\"superscript_a\" style=\"line-height: 1.5;\"><sup><br \/>\na\u00a0 <\/sup>Department of Microbial Biotechnology, Centro Nacional de Biotecnolog\u00eda, Consejo Superior de Investigaciones Cient\u00edficas (CSIC), Campus UAM Cantoblanco, Madrid, Spain<\/span><span style=\"line-height: 1.5;\">\u00a0<\/span><\/div>\n<\/div>\n<p id=\"affiliationlist\" class=\"affilTxt\"><span id=\"superscript_b\"><sup>b\u00a0 <\/sup>Research Institute of Biomedical and Health Sciences, Veterinary Faculty, Universidad de Las Palmas de Gran Canaria (UPGC), Arucas, Las Palmas, Canary Islands, Spain<\/span><br \/>\n<span id=\"superscript_c\"><sup>c\u00a0 <\/sup>Centro de Biolog\u00eda Molecular \u201cSevero Ochoa\u201d (Consejo Superior de Investigaciones Cient\u00edficas\u2014Universidad Aut\u00f3noma de Madrid), Campus UAM Cantoblanco, Madrid, Spain<\/span><\/p>\n<div id=\"viewHideAffilList\"><\/div>\n<h2 class=\"subTitle paddingT7 paddingB10\">Abstract<\/h2>\n<p id=\"recordAbs\" class=\"marginB3\" align=\"justify\">Abnormal levels of fibrinogen (Fib) in blood plasma are associated with several pathological conditions and hence methods for its detection in blood and body fluids are essential. Nanobodies (Nbs) or (VHHs) are single domain antibodies derived from camelids with excellent biophysical and antigen-binding properties, showing great promise in diagnostics and therapy. In this work, we select and characterize high affinity Nbs binding human Fib employing an E. coli cell surface display system based on the fusion of an immune library of VHH domains with the \u03b2-domain of Intimin. Bacteria displaying high-affinity Nbs against Fib were selected using magnetic cell sorting (MACS). Specific binding of the selected clones to Fib was confirmed by flow cytometry of E. coli bacteria, as well as by enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR) with the purified Nbs. E. coli display also provided an excellent estimation of the affinity of the selected Nbs by flow cytometry analysis under equilibrium conditions, with equilibrium constant (K<sub>D<\/sub>) values very similar to those obtained by SPR analysis. Finally, pairwise epitope-scouting studies revealed that the selected Nbs bound distinct epitopes on Fib. The selected Nbs are promising diagnostic tools for determination of human Fib levels.<\/p>\n<\/div>\n","protected":false},"excerpt":{"rendered":"<p>Journal of Biotechnology Volume 234, 20 September 2016, Pages 58-65 \u00a0Salema, V.a,\u00a0L\u00f3pez-Guajardo, A.a, \u00a0Gutierrez, C.b,\u00a0Menc\u00eda, M.c, \u00a0Fern\u00e1ndez, L.\u00c1.a a\u00a0 Department of Microbial Biotechnology, Centro Nacional de Biotecnolog\u00eda, Consejo Superior de Investigaciones Cient\u00edficas (CSIC), Campus UAM Cantoblanco, Madrid, Spain\u00a0 b\u00a0 Research Institute of Biomedical and Health Sciences, Veterinary Faculty, Universidad de Las Palmas de Gran Canaria<\/p>\n","protected":false},"author":1,"featured_media":590,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":[],"categories":[4,1],"tags":[],"_links":{"self":[{"href":"https:\/\/medicinaveterinaria.ulpgc.es\/index.php?rest_route=\/wp\/v2\/posts\/585"}],"collection":[{"href":"https:\/\/medicinaveterinaria.ulpgc.es\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/medicinaveterinaria.ulpgc.es\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/medicinaveterinaria.ulpgc.es\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/medicinaveterinaria.ulpgc.es\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=585"}],"version-history":[{"count":1,"href":"https:\/\/medicinaveterinaria.ulpgc.es\/index.php?rest_route=\/wp\/v2\/posts\/585\/revisions"}],"predecessor-version":[{"id":586,"href":"https:\/\/medicinaveterinaria.ulpgc.es\/index.php?rest_route=\/wp\/v2\/posts\/585\/revisions\/586"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/medicinaveterinaria.ulpgc.es\/index.php?rest_route=\/wp\/v2\/media\/590"}],"wp:attachment":[{"href":"https:\/\/medicinaveterinaria.ulpgc.es\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=585"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/medicinaveterinaria.ulpgc.es\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=585"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/medicinaveterinaria.ulpgc.es\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=585"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}