{"id":1076,"date":"2021-03-29T18:06:48","date_gmt":"2021-03-29T17:06:48","guid":{"rendered":"http:\/\/www.afmb.univ-mrs.fr\/?post_type=facility&#038;p=1076"},"modified":"2025-07-08T16:39:40","modified_gmt":"2025-07-08T15:39:40","slug":"selection-and-production-of-camelid-vhhs-nanobodies","status":"publish","type":"facility","link":"https:\/\/www.afmb.univ-mrs.fr\/en\/facility\/structural-biology\/selection-and-production-of-camelid-vhhs-nanobodies\/","title":{"rendered":"Selection and production of camelid VHHs\/nanobodies"},"content":{"rendered":"\n<div class=\"wp-block-group alignfull pageSubHeaderWrapper topSection has-white-color has-dark-blue-background-color has-text-color has-background\"><div class=\"wp-block-group__inner-container is-layout-flow wp-block-group-is-layout-flow\">\n<ul class=\"wp-block-list\">\n<li><a href=\"#presentation\">Introduction<\/a><\/li>\n<\/ul>\n<\/div><\/div>\n\n\n\n<div class=\"wp-block-columns presentationTeamWrapper is-layout-flex wp-container-core-columns-is-layout-9d6595d7 wp-block-columns-is-layout-flex\" id=\"presentation\">\n<div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow\" style=\"flex-basis:66.66%\">\n<p class=\"is-style-default has-large-font-size\">This <a href=\"https:\/\/www.afmb.univ-mrs.fr\/en\/facility\/structural-biology\/\">PBSIM<\/a> service offers the expertise for selection of camelid VHHs\/nanobodies followed by their production and characterization (SPR, BLI, micro-cal).<\/p>\n\n\n\n<p class=\"is-style-justified\">The use of single-domains from camelid immunoglobulins, termed VHHs or nanobodies (Fig. 1), has seen an increasing growth in recent years. The usefulness of nanobodies in structural biology is now firmly established, as they provide access to new epitopes in concave and hinge regions \u2013 and stabilize them.<\/p>\n\n\n\n<div class=\"wp-block-columns is-layout-flex wp-container-core-columns-is-layout-9d6595d7 wp-block-columns-is-layout-flex\">\n<div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow\">\n<p class=\"is-style-justified\">Remarkable results have been achieved by using nanobodies with flexible multi-domain proteins, large complexes and, last but not least, membrane proteins such as GPCRs or other receptors or channels. The structural biology group at the AFMB lab has been among the pioneers in the work on VHHs and nanobodies.<\/p>\n<\/div>\n\n\n\n<div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow\">\n<figure class=\"wp-block-image size-large\"><a href=\"http:\/\/www.afmb.univ-mrs.fr\/wp-content\/uploads\/2021\/04\/vhhs-Figure-2-d9c57.jpg\"><img loading=\"lazy\" decoding=\"async\" width=\"500\" height=\"324\" src=\"http:\/\/www.afmb.univ-mrs.fr\/wp-content\/uploads\/2021\/04\/vhhs-Figure-2-d9c57.jpg\" alt=\"\" class=\"wp-image-609\" srcset=\"https:\/\/www.afmb.univ-mrs.fr\/wp-content\/uploads\/2021\/04\/vhhs-Figure-2-d9c57.jpg 500w, https:\/\/www.afmb.univ-mrs.fr\/wp-content\/uploads\/2021\/04\/vhhs-Figure-2-d9c57-300x194.jpg 300w\" sizes=\"auto, (max-width: 500px) 100vw, 500px\" \/><\/a><\/figure>\n<\/div>\n<\/div>\n\n\n\n<p class=\"is-style-justified\">This tecnological facility offers immunization of llamas (<em>Lama glama glama<\/em>) or camels (<em>Camelus dromedarius<\/em>) in view to obtain target specific libraries, panning of these libraries by phage display from which several vHHs\/nanobodies are cloned (service 1), expressed (service 2) and characterized (service 3) (Fig. 2). We also propose our \u201cnaive\u201d library obtained by gathering the lymphocytes of non-immunized animals \u2013 in our case, 3 camels and 4 llamas. Randomizing one of the three CDRs (generally CDR3) from a selected VHH\/nanobody followed by a new panning cycle (service 4) leads to sub-nanomolar affinities.<\/p>\n\n\n\n<div class=\"wp-block-columns tableau is-layout-flex wp-container-core-columns-is-layout-9d6595d7 wp-block-columns-is-layout-flex\">\n<div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow\">\n<div id=\"network\" class=\"wp-block-group\"><div class=\"wp-block-group__inner-container is-layout-flow wp-block-group-is-layout-flow\">\n<h6 class=\"wp-block-heading is-style-default has-orange-color has-text-color\" id=\"accessibility\">Accessibility<\/h6>\n\n\n\n<p>Academia and industries<\/p>\n<\/div><\/div>\n<\/div>\n\n\n\n<div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow\">\n<div id=\"funding\" class=\"wp-block-group\"><div class=\"wp-block-group__inner-container is-layout-flow wp-block-group-is-layout-flow\">\n<h6 class=\"wp-block-heading is-style-default has-orange-color has-text-color\" id=\"cost\">Cost<\/h6>\n\n\n\n<p>To be defined from the service requested.<\/p>\n<\/div><\/div>\n<\/div>\n<\/div>\n\n\n\n<div class=\"wp-block-columns tableau is-layout-flex wp-container-core-columns-is-layout-9d6595d7 wp-block-columns-is-layout-flex\">\n<div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow\">\n<div id=\"network\" class=\"wp-block-group\"><div class=\"wp-block-group__inner-container is-layout-flow wp-block-group-is-layout-flow\">\n<h6 class=\"wp-block-heading is-style-default has-orange-color has-text-color\" id=\"comment-faire-une-demande\">Comment faire une demande ?<\/h6>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>Contact :<\/strong> <a href=\"https:\/\/www.afmb.univ-mrs.fr\/en\/member\/anais-gaubert\/\" target=\"_blank\" rel=\"noreferrer noopener\">Ana\u00efs Gaubert<\/a><\/li>\n<\/ul>\n<\/div><\/div>\n<\/div>\n<\/div>\n\n\n\n<div class=\"wp-block-columns tableau is-layout-flex wp-container-core-columns-is-layout-9d6595d7 wp-block-columns-is-layout-flex\">\n<div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow\">\n<div id=\"network\" class=\"wp-block-group is-vertical is-layout-flex wp-container-core-group-is-layout-8cf370e7 wp-block-group-is-layout-flex\">\n<h6 class=\"wp-block-heading is-style-default has-orange-color has-text-color\" id=\"specific-equipement\">Specific equipement<\/h6>\n\n\n\n<ul class=\"wp-block-list\">\n<li>FPLC \u00c4kta Purifier and Xpress (GE Heathcare)<\/li>\n\n\n\n<li>Microcalorimetry: MicroCal iTC200 (GE Heathcare)<\/li>\n\n\n\n<li>Microscale thermophoresis: MST Monolith NT.115 (NanoTemper Technologies GmbH)<\/li>\n\n\n\n<li>Bio-layer interferometry: Octet RED96 system and Blitz system (Pall ForteBio LLC)<\/li>\n\n\n\n<li>Size exclusion chromatography \/ multi-angle light scattering \/ refractometry: HPLC Alliance (Waters) \/ Minidawn Treos (Wyatt) \/ Optilab rEX (Wyatt)<\/li>\n<\/ul>\n\n\n\n<p><\/p>\n\n\n\n<h2 class=\"wp-block-heading\">Members<\/h2>\n\n\n\n<div class=\"wp-block-group is-content-justification-center is-nowrap is-layout-flex wp-container-core-group-is-layout-94bc23d7 wp-block-group-is-layout-flex\"><div class=\"lazyblock-member-block-Z17kYWw wp-block-lazyblock-member-block\"><div class=\"memberInList fadeIn\">\n    <a class=\"itemLink\" href=\"https:\/\/www.afmb.univ-mrs.fr\/en\/member\/anais-gaubert\/\">\n        <div class=\"imgWrapper\">\n                                    <img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/www.afmb.univ-mrs.fr\/wp-content\/uploads\/2021\/07\/Anais-Gaubert-scaled-e1655373789620-500x500.jpg\" width=\"500\" height=\"500\" alt=\"\" class=\"lazyload\" \/>\n                    <\/div>\n        <div class=\"memberInfos\">\n            <p class=\"memberInListName\">Ana\u00efs Gaubert<\/p>\n                        <div class=\"memberInListExcerpt\">\n            <p>Engineer<\/p>\n            <\/div>\n                                <\/div>\n    <\/a>\n<\/div><\/div><\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n\n\n\n<div class=\"wp-block-column columnStickyWrapper is-layout-flow wp-block-column-is-layout-flow\" style=\"flex-basis:33.33%\">\n<div class=\"wp-block-group stickyColumnGroup\"><div class=\"wp-block-group__inner-container is-layout-flow wp-block-group-is-layout-flow\"><div class=\"singleTeamLeaders clearfix\"><div class=\"singleTeamContactWrapper\"><h5>Team contacts<\/h5><ul><li><a href=\"https:\/\/frisbi.eu\/centers\/afmb\/\" target=\"_blank\"><i class=\"fas fa-link\"><\/i><\/a><\/li><li><a href=\"mailto:anais.gaubert@univ-amu.fr\"><i class=\"far fa-envelope\"><\/i><\/a><\/li><li class=\"phoneIconWrapper\"><a class=\"phoneIconLink\"><i class=\"fas fa-tty\"><\/i><\/a><div class=\"phoneNumber\"><a href=\"tel:+33 (0)4 13 94 95 10\">+33 (0)4 13 94 95 10<\/a><\/div><\/li><\/ul><\/div><\/div>\n<\/div><\/div>\n<\/div>\n<\/div>\n\n\n\n<div class=\"wp-block-group accordeon\"><div class=\"wp-block-group__inner-container is-layout-flow wp-block-group-is-layout-flow\">\n<h4 class=\"wp-block-heading accordeonTitle\" id=\"publications\">Publications <\/h4>\n\n\n\n<div class=\"wp-block-group accordeonContent\"><div class=\"wp-block-group__inner-container is-layout-flow wp-block-group-is-layout-flow\">\n<ul class=\"wp-block-list\">\n<li>Durand E, Nguyen VS, Zoued A, Logger L, P\u00e9hau-Arnaudet G, Aschtgen M-S, Spinelli S, Desmyter A, Bardiaux B, Dujeancourt A, Roussel A, Cambillau C, Cascales E, Fronzes R. Biogenesis and structure of the bacterial Type VI secretion membrane core complex. <strong>Nature<\/strong> 2015, 300:555-560 [doi: 10.1038\/nature14667]<\/li>\n\n\n\n<li>Nguyen VS, Logger L, Spinelli S, Desmyter A, Le TTH, Kellenberger C, Douzi B, Durand E, Roussel A, Cascales E, Cambillau C. Inhibition of type VI secretion by an anti-TssM llama nanobody, <strong>PloS-ONE<\/strong> 2015, 10(3):e0122187. [doi:10.1371\/journal.pone.0122187]<\/li>\n\n\n\n<li>Nguyen VS, Spinelli S, Desmyter A, Le TTH, Kellenberger C, Cascales E, Cambillau C, Roussel A. Production, crystallization and X-ray diffraction analysis of a complex between a fragment of the TssM T6SS protein and a Camelid nanobody, <strong>Acta Crystallogr. F<\/strong> 2015, F71:266-271<\/li>\n\n\n\n<li>Desmyter A, Spinelli S, Roussel A, Cambillau C. Camelid nanobodies: killing two birds with one stone, <strong>Curr. Opin. Struct. Biol.<\/strong>, 2015, 32C:1\u20138 [doi: 10.1016\/j.sbi.2015.01.001]<\/li>\n\n\n\n<li>Hassaine G, Deluz C, Grasso L, Wyss R, Tol MB, Hovius R, Graff A, Stahlberg H, Tomizaki T, Desmyter A, Moreau C, Li XD, Poitevin F, Vogel H, Nury H. X-ray structure of the mouse serotonin 5-HT3 receptor. <strong>Nature<\/strong> 2014, 512:276-281.<\/li>\n<\/ul>\n<\/div><\/div>\n<\/div><\/div>\n","protected":false},"excerpt":{"rendered":"<p>This service offers a set of expertises ranging from generation and selection of camelid nanobodies up to their production and functional characterization (SPR, BLI, micro-cal).<\/p>\n","protected":false},"author":1,"featured_media":0,"parent":1060,"menu_order":3,"template":"","class_list":["post-1076","facility","type-facility","status-publish","hentry","entry"],"_links":{"self":[{"href":"https:\/\/www.afmb.univ-mrs.fr\/en\/wp-json\/wp\/v2\/facility\/1076","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.afmb.univ-mrs.fr\/en\/wp-json\/wp\/v2\/facility"}],"about":[{"href":"https:\/\/www.afmb.univ-mrs.fr\/en\/wp-json\/wp\/v2\/types\/facility"}],"author":[{"embeddable":true,"href":"https:\/\/www.afmb.univ-mrs.fr\/en\/wp-json\/wp\/v2\/users\/1"}],"version-history":[{"count":26,"href":"https:\/\/www.afmb.univ-mrs.fr\/en\/wp-json\/wp\/v2\/facility\/1076\/revisions"}],"predecessor-version":[{"id":11183,"href":"https:\/\/www.afmb.univ-mrs.fr\/en\/wp-json\/wp\/v2\/facility\/1076\/revisions\/11183"}],"up":[{"embeddable":true,"href":"https:\/\/www.afmb.univ-mrs.fr\/en\/wp-json\/wp\/v2\/facility\/1060"}],"wp:attachment":[{"href":"https:\/\/www.afmb.univ-mrs.fr\/en\/wp-json\/wp\/v2\/media?parent=1076"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}