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A recombinant dromedary antibody fragment (VHH or nanobody) directed against human Duffy antigen receptor for chemokines

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Abstract

Fy blood group antigens are carried by the Duffy antigen receptor for chemokines (DARC), a red cells receptor for Plasmodium vivax broadly implicated in human health and diseases. Recombinant VHHs, or nanobodies, the smallest intact antigen binding fragment derivative from the heavy chain-only antibodies present in camelids, were prepared from a dromedary immunized against DARC N-terminal extracellular domain and selected for DARC binding. A described VHH, CA52, does recognize native DARC on cells. It inhibits P. vivax invasion of erythrocytes and displaces interleukin-8 bound to DARC. The targeted epitope overlaps the well-defined DARC Fy6 epitope. K D of CA52–DARC equilibrium is sub-nanomolar, hence ideal to develop diagnostic or therapeutic compounds. Immunocapture by immobilized CA52 yielded highly purified DARC from engineered K562 cells. This first report on a VHH with specificity for a red blood cell protein exemplifies VHHs’ potentialities to target, to purify, and to modulate the function of cellular markers.

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Abbreviations

DARC:

Duffy antigen receptor for chemokines

ECD1:

First extracellular domain

CNRGS:

Centre National de Reference pour les Groupes Sanguins

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Acknowledgments

The deepest gratitude is due to Inge Mestdagh and Jan van Gompel for their generous help and their patient teaching during library construction and screening. D.S. was supported by scholarship from French Embassy in Warsaw and from the Flemish government. The project was supported by grant no. N N302 118835 from the Ministry of Science and Higher Education of Poland and in part by a Polonium Partenariat Hubert Curien grant. Authors thank Dr. Bach Nga Pham and Eliane Vera at CNRGS for making available phenotyped human blood, Dr. Christophe Tournamille for the recombinant K562 cells, and Prof. Donald Engelman for the plasmid used for some of the E. coli protein productions.

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Authors and Affiliations

  1. INSERM, UMR_S 665, 75015, Paris, France

    Dorota Smolarek, Claude Hattab, Sylvie Cochet, Alexandre G. de Brevern, Julien Picot, Yves Colin, Caroline Le Van Kim & Olivier Bertrand

  2. Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, Poland

    Dorota Smolarek, Magdalena Grodecka, Kazimiera Wasniowska & Marcin Czerwinski

  3. Institut National de la Transfusion Sanguine, 75015, Paris, France

    Dorota Smolarek, Claude Hattab, Sylvie Cochet, Alexandre G. de Brevern, Julien Picot, Yves Colin, Caroline Le Van Kim & Olivier Bertrand

  4. Université Paris7-Denis Diderot, 75013, Paris, France

    Dorota Smolarek, Claude Hattab, Sylvie Cochet, Alexandre G. de Brevern, Julien Picot, Yves Colin, Caroline Le Van Kim & Olivier Bertrand

  5. Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, 1050, Brussels, Belgium

    Gholamreza Hassanzadeh-Ghassabeh & Serge Muyldermans

  6. Department of Molecular and Cellular Interactions, VIB, Brussels, Belgium

    Gholamreza Hassanzadeh-Ghassabeh & Serge Muyldermans

  7. Department of Animal Medicine and Surgery, Veterinary Faculty, University of Las Palmas, Las Palmas, Spain

    Carlos Gutiérrez

  8. Department of Pathobiology, Faculty of Science, Mahidol University, Bangkok, Thailand

    Rachanee Udomsangpetch

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Smolarek, D., Hattab, C., Hassanzadeh-Ghassabeh, G. et al. A recombinant dromedary antibody fragment (VHH or nanobody) directed against human Duffy antigen receptor for chemokines. Cell. Mol. Life Sci. 67, 3371–3387 (2010). https://doi.org/10.1007/s00018-010-0387-6

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