ǂa ǂuniversal synthetic library of humanized nanobodies providing highly functional antibodies and intrabodies
Sandrine Moutel (Author), Nicolas Bery (Author), Virginie Bernard (Author), Laura Keller (Author), Emilie Lemesre (Author), Ario De Marco (Author), Laetitia Ligat (Author), Jean-Christophe Rain (Author), Gilles Favre (Author), Aurelien Olichon (Author), Franck Perez (Author)

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;

Data

Language: English
Year of publishing:
Typology: 1.01 - Original Scientific Article
Organization: UNG - University of Nova Gorica
UDC: 602
COBISS: 4478971 Link will open in a new window
ISSN: 2050-084X
Views: 4272
Downloads: 238
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Other data

URN: URN:SI:UNG
Type (COBISS): Not categorized
Pages: str. 1-31
Volume: Vol. 5
Issue: ǂVol. ǂ5
Chronology: 19. Jul. 2016
DOI: 10.7554/eLife.16228
ID: 9161126
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