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

  1. Sandrine Moutel
  2. Nicolas Bery
  3. Virginie Bernard
  4. Laura Keller
  5. Emilie Lemesre
  6. Ario de Marco
  7. Laetitia Ligat
  8. Jean-Christophe Rain
  9. Gilles Favre
  10. Aurélien Olichon  Is a corresponding author
  11. Franck Perez  Is a corresponding author
  1. Institut Curie, France
  2. Inserm, UMR 1037-CRCT, France
  3. CRCT, France
  4. Hybrigenics Service SA, 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.

Article and author information

Author details

  1. Sandrine Moutel

    Institut Curie, Paris, France
    Competing interests
    Sandrine Moutel, Co-inventor on a patent application (filled under ref: WO/2015/063331) that covers the hs2dAb scaffold and the commercial use of the library. The library has been licensed to Hybrigenics Service SA, which will perform screens on a fee-for-service basis.A consultant for Hybrigenics Service SA.
  2. Nicolas Bery

    Inserm, UMR 1037-CRCT, Toulouse, France
    Competing interests
    No competing interests declared.
  3. Virginie Bernard

    Institut Curie, Paris, France
    Competing interests
    No competing interests declared.
  4. Laura Keller

    Inserm, UMR 1037-CRCT, Toulouse, France
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1786-9760
  5. Emilie Lemesre

    Institut Curie, Paris, France
    Competing interests
    No competing interests declared.
  6. Ario de Marco

    Institut Curie, Paris, France
    Competing interests
    No competing interests declared.
  7. Laetitia Ligat

    plateau de protéomique, CRCT, Toulouse, France
    Competing interests
    No competing interests declared.
  8. Jean-Christophe Rain

    Hybrigenics Service SA, Paris, France
    Competing interests
    Jean-Christophe Rain, Employed by, and a stockholder in, Hybrigenics Service SA.
  9. Gilles Favre

    Inserm, UMR 1037-CRCT, Toulouse, France
    Competing interests
    No competing interests declared.
  10. Aurélien Olichon

    Inserm, UMR 1037-CRCT, Toulouse, France
    For correspondence
    aurelien.olichon@inserm.fr
    Competing interests
    Aurélien Olichon, Co-inventor on a patent application (filled under ref: WO/2015/063331) that covers the hs2dAb scaffold and the commercial use of the library. The library has been licensed to Hybrigenics Service SA, which will perform screens on a fee-for-service basis.
  11. Franck Perez

    Institut Curie, Paris, France
    For correspondence
    Franck.Perez@curie.fr
    Competing interests
    Franck Perez, Co-inventor on a patent application (filled under ref: WO/2015/063331) that covers the hs2dAb scaffold and the commercial use of the library. The library has been licensed to Hybrigenics Service SA, which will perform screens on a fee-for-service basis.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9129-9401

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.

Reviewing Editor

  1. Anna Akhmanova, Utrecht University, Netherlands

Version history

  1. Received: March 20, 2016
  2. Accepted: July 18, 2016
  3. Accepted Manuscript published: July 19, 2016 (version 1)
  4. Version of Record published: August 15, 2016 (version 2)

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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  1. Sandrine Moutel
  2. Nicolas Bery
  3. Virginie Bernard
  4. Laura Keller
  5. Emilie Lemesre
  6. Ario de Marco
  7. Laetitia Ligat
  8. Jean-Christophe Rain
  9. Gilles Favre
  10. Aurélien Olichon
  11. Franck Perez
(2016)
NaLi-H1: A universal synthetic library of humanized nanobodies providing highly functional antibodies and intrabodies
eLife 5:e16228.
https://doi.org/10.7554/eLife.16228

Share this article

https://doi.org/10.7554/eLife.16228

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