title: NaLi-H1: A universal synthetic library of humanized nanobodies providing highly functional antibodies and intrabodies.
abstract: In vitro selection of antibodies allows to obtain highly functional binders, rapidly and at lower cost. Here, we describe the first fully synthetic phage display library of humanized llama single domain antibody (NaLi-H1: Nanobody Library Humanized 1). Based on a humanized synthetic single domain antibody (hs2dAb) scaffold optimized for intracellular stability, the highly diverse library provides high affinity binders without animal immunization. NaLi-H1 was screened following several selection schemes against various targets (Fluorescent proteins, actin, tubulin, p53, HP1). Conformation antibodies against active RHO GTPase were also obtained. Selected hs2dAb were used in various immunoassays and were often found to be functional intrabodies, enabling tracking or inhibition of endogenous targets. Functionalization of intrabodies allowed specific protein knockdown in living cells. Finally, direct selection against the surface of tumor cells produced hs2dAb directed against tumor-specific antigens further highlighting the potential use of this library for therapeutic applications.
authors.affiliation: Institut Curie, PSL Research University, Paris, France.|CNRS UMR144, Paris, France.|Translational Research Department, Institut Curie, Paris, France.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Moutel
authors.affiliation: Inserm, UMR 1037-CRCT, Toulouse, France.|Faculté des Sciences Pharmaceutiques, Université Toulouse III-Paul Sabatier, Toulouse, France.
authors.forename: Nicolas
authors.identifier: 0000-0002-2643-3897
authors.initials: N
authors.lastname: Bery
authors.affiliation: Institut Curie, PSL Research University, Paris, France.
authors.forename: Virginie
authors.initials: V
authors.lastname: Bernard
authors.affiliation: Inserm, UMR 1037-CRCT, Toulouse, France.|Faculté des Sciences Pharmaceutiques, Université Toulouse III-Paul Sabatier, Toulouse, France.|Institut Claudius Regaud, Toulouse, France.
authors.forename: Laura
authors.identifier: 0000-0002-1786-9760
authors.initials: L
authors.lastname: Keller
authors.affiliation: Institut Curie, PSL Research University, Paris, France.|CNRS UMR144, Paris, France.
authors.forename: Emilie
authors.initials: E
authors.lastname: Lemesre
authors.affiliation: Institut Curie, PSL Research University, Paris, France.
authors.forename: Ario
authors.identifier: 0000-0001-7729-819X
authors.initials: A
authors.lastname: de Marco
authors.affiliation: Le Pôle Technologique du Centre de Recherches en Cancérologie de Toulouse, plateau de protéomique, Toulouse, France.
authors.forename: Laetitia
authors.initials: L
authors.lastname: Ligat
authors.affiliation: Hybrigenics Service, Paris, France.
authors.forename: Jean-Christophe
authors.initials: JC
authors.lastname: Rain
authors.affiliation: Inserm, UMR 1037-CRCT, Toulouse, France.|Faculté des Sciences Pharmaceutiques, Université Toulouse III-Paul Sabatier, Toulouse, France.|Institut Claudius Regaud, Toulouse, France.
authors.forename: Gilles
authors.identifier: 0000-0002-2344-1883
authors.initials: G
authors.lastname: Favre
authors.affiliation: Inserm, UMR 1037-CRCT, Toulouse, France.|Faculté des Sciences Pharmaceutiques, Université Toulouse III-Paul Sabatier, Toulouse, France.
authors.forename: Aurélien
authors.initials: A
authors.lastname: Olichon
authors.affiliation: Institut Curie, PSL Research University, Paris, France.|CNRS UMR144, Paris, France.
authors.forename: Franck
authors.identifier: 0000-0002-9129-9401
authors.initials: F
authors.lastname: Perez
mesh_terms: D000818:Animals; D061067:Antibodies, Monoclonal, Humanized; D002161:Camelids, New World; D006801:Humans; D008967:Molecular Biology / Q000379:methods*; D019151:Peptide Library; D061905:Single-Domain Antibodies
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D019151:Peptide Library; D061905:Single-Domain Antibodies
keywords: E. coli; cell biology; human; immunology; intrabodies; mouse; phage display; recombinant antibodies; synthetic library