Affordable Access

deepdyve-link
Publisher Website

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

Authors
  • Moutel, Sandrine1, 2, 3
  • Bery, Nicolas4, 5
  • Bernard, Virginie1
  • Keller, Laura4, 5, 6
  • Lemesre, Emilie1, 2
  • de Marco, Ario1
  • Ligat, Laetitia7
  • Rain, Jean-Christophe8
  • Favre, Gilles4, 5, 6
  • Olichon, Aurélien4, 5
  • Perez, Franck1, 2
  • 1 Institut Curie, PSL Research University, Paris, France. , (France)
  • 2 CNRS UMR144, Paris, France. , (France)
  • 3 Translational Research Department, Institut Curie, Paris, France. , (France)
  • 4 Inserm, UMR 1037-CRCT, Toulouse, France. , (France)
  • 5 Faculté des Sciences Pharmaceutiques, Université Toulouse III-Paul Sabatier, Toulouse, France. , (France)
  • 6 Institut Claudius Regaud, Toulouse, France. , (France)
  • 7 Le Pôle Technologique du Centre de Recherches en Cancérologie de Toulouse, plateau de protéomique, Toulouse, France. , (France)
  • 8 Hybrigenics Service, Paris, France. , (France)
Type
Published Article
Journal
eLife
Publisher
"eLife Sciences Organisation, Ltd."
Publication Date
Jul 19, 2016
Volume
5
Identifiers
DOI: 10.7554/eLife.16228
PMID: 27434673
Source
Medline
Keywords
License
Unknown

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.

Report this publication

Statistics

Seen <100 times