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A recombinant dromedary antibody fragment (VHH or nanobody) directed against human Duffy antigen receptor for chemokines |
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| Authors: | Dorota Smolarek Claude Hattab Gholamreza Hassanzadeh-Ghassabeh Sylvie Cochet Carlos Guti��rrez Alexandre G de Brevern Rachanee Udomsangpetch Julien Picot Magdalena Grodecka Kazimiera Wasniowska Serge Muyldermans Yves Colin Caroline Le Van Kim Marcin Czerwinski Olivier Bertrand |
| Institution: | 1. INSERM, UMR_S 665, 75015, Paris, France 2. Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, Poland 3. Institut National de la Transfusion Sanguine, 75015, Paris, France 4. Universit?? Paris7-Denis Diderot, 75013, Paris, France 5. Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, 1050, Brussels, Belgium 6. Department of Molecular and Cellular Interactions, VIB, Brussels, Belgium 7. Department of Animal Medicine and Surgery, Veterinary Faculty, University of Las Palmas, Las Palmas, Spain 8. Department of Pathobiology, Faculty of Science, Mahidol University, Bangkok, Thailand |
| 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?CDARC 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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