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Title:NaLi-H1: A universal synthetic library of humanized nanobodies providing highly functional antibodies and intrabodies
Authors:ID Moutel, Sandrine, Institut Curie (Author)
ID Bery, Nicolas, Inserm (Author)
ID Bernard, Virginie, Institut Curie (Author)
ID Keller, Laura, Inserm (Author)
ID Lemesre, Emilie, Institut Curie (Author)
ID De Marco, Ario, Institut Curie (Author)
ID Ligat, Laetitia, Cancerologie Toulouse (Author)
ID Rain, Jean-Christophe, Hybrigenics (Author)
ID Fevre, Gilles, Inserm (Author)
ID Olichon, Aurelien, Inserm (Author)
ID Perez, Franck, Institut Curie (Author)
Files:.pdf Synthetic_Library.pdf (4,32 MB)
MD5: 1583F8AAB5ED123A90BE47D4323A8038
Work type:Not categorized
Typology:1.01 - Original Scientific Article
Organization:UNG - University of Nova Gorica
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.
Keywords:nanobodies, synthetic phage display library, in vitro panning
Publication version:Version of Record
Year of publishing:2016
Number of pages:31
PID:20.500.12556/RUNG-2536 New window
COBISS.SI-ID:4478971 New window
DOI:10.7554/eLife.16228 New window
Publication date in RUNG:17.08.2016
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Shortened title:eLIFE
Year of publishing:2016