A recombinant dromedary antibody fragment (VHH or nanobody) directed against human Duffy antigen receptor for chemokines.
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.
Smolarek
Dorota
D
INSERM, UMR_S 665, 75015 Paris, France.
Hattab
Claude
C
Hassanzadeh-Ghassabeh
Gholamreza
G
Cochet
Sylvie
S
Gutiérrez
Carlos
C
de Brevern
Alexandre G
AG
Udomsangpetch
Rachanee
R
Picot
Julien
J
Grodecka
Magdalena
M
Wasniowska
Kazimiera
K
Muyldermans
Serge
S
Colin
Yves
Y
Le Van Kim
Caroline
C
Czerwinski
Marcin
M
Bertrand
Olivier
O
D000818:Animals; D001789:Blood Group Antigens / Q000276:immunology / Q000378:metabolism; D002162:Camelus / Q000276:immunology / Q000378:metabolism; D002352:Carrier Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D018925:Chemokines / Q000276:immunology / Q000378:metabolism*; D004375:Duffy Blood-Group System; D004912:Erythrocytes / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007136:Immunoglobulins / Q000276:immunology / Q000378:metabolism; D016209:Interleukin-8 / Q000276:immunology / Q000378:metabolism; D010966:Plasmodium vivax / Q000276:immunology / Q000378:metabolism; D011946:Receptors, Antigen / Q000276:immunology / Q000378:metabolism; D011956:Receptors, Cell Surface; D019707:Receptors, Chemokine / Q000276:immunology / Q000378:metabolism; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism
C473745:ACKR1 protein, human; D001789:Blood Group Antigens; D002352:Carrier Proteins; D018925:Chemokines; D004375:Duffy Blood-Group System; D007136:Immunoglobulins; D016209:Interleukin-8; D011946:Receptors, Antigen; D011956:Receptors, Cell Surface; D019707:Receptors, Chemokine; D011994:Recombinant Proteins
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