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Elife. 2016 Jul 19;5. pii: e16228. doi: 10.7554/eLife.16228.

NaLi-H1: A universal synthetic library of humanized nanobodies providing highly functional antibodies and intrabodies.

Author information

1
Institut Curie, PSL Research University, Paris, France.
2
CNRS UMR144, Paris, France.
3
Translational Research Department, Institut Curie, Paris, France.
4
Inserm, UMR 1037-CRCT, Toulouse, France.
5
Faculté des Sciences Pharmaceutiques, Université Toulouse III-Paul Sabatier, Toulouse, France.
6
Institut Claudius Regaud, Toulouse, France.
7
Le Pôle Technologique du Centre de Recherches en Cancérologie de Toulouse, plateau de protéomique, Toulouse, France.
8
Hybrigenics Service, Paris, France.

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:

E. coli; cell biology; human; immunology; intrabodies; mouse; phage display; recombinant antibodies; synthetic library

PMID:
27434673
PMCID:
PMC4985285
DOI:
10.7554/eLife.16228
[Indexed for MEDLINE]
Free PMC Article

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