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.
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Author details
Funding
Institut Curie
- Sandrine Moutel
- Virginie Bernard
- Ario de Marco
- Franck Perez
Centre National de la Recherche Scientifique
- Sandrine Moutel
- Franck Perez
Institut National de la Santé et de la Recherche Médicale
- Nicolas Bery
- Laura Keller
- Laetitia Ligat
- Gilles Favre
- Aurélien Olichon
Agence Nationale de la Recherche (ANR-09-BIOT-05)
- Sandrine Moutel
- Jean-Christophe Rain
- Franck Perez
Institut National de la Santé et de la Recherche Médicale (ITS-201103)
- Ario de Marco
- Franck Perez
Fondation pour la Recherche Médicale (DEQ20120323723)
- Franck Perez
Groupe de Recherche of the Claudius Regaud Institute
- Gilles Favre
- Aurélien Olichon
LABEX CellTisPhyBio (11-LBX-0038)
- Sandrine Moutel
- Virginie Bernard
- Ario de Marco
- Franck Perez
IDEX Paris Sciences Lettres (ANR-10-IDEX-0001-02 PSL)
- Sandrine Moutel
- Virginie Bernard
- Ario de Marco
- Franck Perez
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2016, Moutel et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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