Skip Navigation


PEDS Advance Access originally published online on December 11, 2007
Protein Engineering Design and Selection 2008 21(1):1-10; doi:10.1093/protein/gzm064
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
21/1/1    most recent
gzm064v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Google Scholar
Right arrow Articles by Behar, G.
Right arrow Articles by Baty, D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Behar, G.
Right arrow Articles by Baty, D.
Related Collections
Right arrow 2008
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2007. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Isolation and characterization of anti-Fc{gamma}RIII (CD16) llama single-domain antibodies that activate natural killer cells

Ghislaine Behar1, Sophie Sibéril2,3,4, Agnès Groulet1, Patrick Chames1, Martine Pugnière5, Charlotte Boix2,3,4, Catherine Sautès-Fridman2,3,4, Jean-Luc Teillaud2,3,4,6,7 and Daniel Baty1,6,7

1 Laboratoire d’Ingénierie des Systèmes Macromoléculaires, CNRS, UPR9027, GDR2352, 31 chemin Joseph Aiguier, F-13402 Marseille Cedex 20, France 2 INSERM, U872, CNRS GDR2352, 15 rue de l’école de médecine, F-75006 Paris, France 3Centre de Recherche des Cordeliers, Université Pierre et Marie Curie – Paris 6 UMRS872, Paris F-75006, France 4 Université Paris Descartes, UMRS872, Paris, F-75006 France 5 Centre de Pharmacologie et Innovation dans le Diabète, Faculté de Pharmacie, CNRS, UMR5232, 15 Avenue Charles Flahault, BP 14491, F-34093 Montpellier Cedex 5, GDR2352, France

7 To whom correspondance should be addressed. E-mail: baty{at}ibsm.cnrs-mrs.fr (D.B.)/ jean-luc.teillaud{at}crc.jussieu.fr (J.-L.T.)

Fc{gamma}RIII (CD16) plays an important role in the anti-tumor effects of therapeutic antibodies. Bi-specific antibodies (bsAbs) targeting Fc{gamma}RIII represent a powerful alternative to the recruitment of the receptor via the Fc fragment, but are not efficiently produced. Single-domain antibodies (sdAbs) endowed with many valuable structural features might help to bypass this problem. In the present work, we have isolated anti-Fc{gamma}RIII sdAbs (C21 and C28) from a phage library generated from a llama immunized with Fc{gamma}RIIIB extra-cellular domains. These sdAbs bind Fc{gamma}RIIIA+ NK cells and Fc{gamma}RIIIB+ polymorphonuclear cells, but not Fc{gamma}RI+ or Fc{gamma}RII+ cells, as detected by indirect immunofluorescence. Competition experiments showed that C21 and C28 sdAbs bind different Fc{gamma}RIII epitopes, with C21 recognizing a linear and C28 a conformational epitope of the receptor. Surface plasmon resonance experiments showed that C21 and C28 sdAbs bind Fc{gamma}RIII with a KD in the 10 and 80 nM range, respectively. Importantly, the engagement by both molecules of Fc{gamma}RIIIA expressed by transfected Jurkat T cells or by NK cells derived from peripheral blood induced a strong IL-2 and IFN-{gamma} production, respectively. These anti-Fc{gamma}RIII sdAbs represent versatile tools for generating bsAbs under various formats, able to recruit Fc{gamma}RIII killer cells to target and destroy tumor cells.

Keywords: CD16/Fc{gamma}RIII/llama/phage display/single-domain antibodies

Received June 26, 2007; revised September 4, 2007; accepted October 8, 2007.

6 J.-L.T. and D.B. are senior co-authors.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.