title: A recombinant dromedary antibody fragment (VHH or nanobody) directed against human Duffy antigen receptor for chemokines.
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.
authors.affiliation: INSERM, UMR_S 665, 75015 Paris, France.
authors.forename: Dorota
authors.initials: D
authors.lastname: Smolarek
authors.forename: Claude
authors.initials: C
authors.lastname: Hattab
authors.forename: Gholamreza
authors.initials: G
authors.lastname: Hassanzadeh-Ghassabeh
authors.forename: Sylvie
authors.initials: S
authors.lastname: Cochet
authors.forename: Carlos
authors.initials: C
authors.lastname: Gutiérrez
authors.forename: Alexandre G
authors.initials: AG
authors.lastname: de Brevern
authors.forename: Rachanee
authors.initials: R
authors.lastname: Udomsangpetch
authors.forename: Julien
authors.initials: J
authors.lastname: Picot
authors.forename: Magdalena
authors.initials: M
authors.lastname: Grodecka
authors.forename: Kazimiera
authors.initials: K
authors.lastname: Wasniowska
authors.forename: Serge
authors.initials: S
authors.lastname: Muyldermans
authors.forename: Yves
authors.initials: Y
authors.lastname: Colin
authors.forename: Caroline
authors.initials: C
authors.lastname: Le Van Kim
authors.forename: Marcin
authors.initials: M
authors.lastname: Czerwinski
authors.forename: Olivier
authors.initials: O
authors.lastname: Bertrand
mesh_terms: 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
chemical_list: 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
references.citation: Methods Enzymol. 2009;463:631-45
references.pmid: 19892196
references.citation: N Engl J Med. 1976 Aug 5;295(6):302-4
references.pmid: 778616
references.citation: Cell Host Microbe. 2009 May 8;5(5):408-10; author reply 418-9
references.pmid: 19454339
references.citation: Nucleic Acids Res. 1997 Sep 1;25(17):3389-402
references.pmid: 9254694
references.citation: Mol Microbiol. 2005 Mar;55(5):1413-22
references.pmid: 15720550
references.citation: Cell Host Microbe. 2009 May 8;5(5):413-5; author reply 418-9
references.pmid: 19454341
references.citation: Proc Natl Acad Sci U S A. 2006 Mar 21;103(12):4586-91
references.pmid: 16537393
references.citation: Mol Pharmacol. 2008 May;73(5):1362-70
references.pmid: 18230715
references.citation: Cell Host Microbe. 2009 May 8;5(5):415-7; author reply 418-9
references.pmid: 19454342
references.citation: Mol Biochem Parasitol. 2005 Nov;144(1):100-3
references.pmid: 16024106
references.citation: Nature. 1993 Jun 3;363(6428):446-8
references.pmid: 8502296
references.citation: EMBO J. 2000 Mar 1;19(5):921-30
references.pmid: 10698934
references.citation: Oncogene. 2006 Nov 16;25(54):7201-11
references.pmid: 16785997
references.citation: Blood. 2000 May 15;95(10):3032-43
references.pmid: 10807766
references.citation: Nat Med. 2006 May;12(5):580-4
references.pmid: 16604085
references.citation: Blood. 2009 Sep 24;114(13):2783-92
references.pmid: 19620399
references.citation: PLoS One. 2010 Jan 28;5(1):e8921
references.pmid: 20126667
references.citation: Biochem J. 2004 Apr 15;379(Pt 2):263-72
references.pmid: 14723600
references.citation: Semin Hematol. 2000 Apr;37(2):122-9
references.pmid: 10791881
references.citation: J Mol Biol. 1993 Dec 5;234(3):779-815
references.pmid: 8254673
references.citation: J Exp Med. 1987 Sep 1;166(3):776-85
references.pmid: 2442291
references.citation: Trends Pharmacol Sci. 2006 Jan;27(1):41-7
references.pmid: 16310864
references.citation: Biochem J. 2009 Nov 11;424(2):263-72
references.pmid: 19732033
references.citation: Biochem Soc Trans. 2006 Dec;34(Pt 6):1005-8
references.pmid: 17073738
references.citation: Protein Sci. 2008 Jul;17(7):1175-87
references.pmid: 18445622
references.citation: Mol Cell Biol. 1988 May;8(5):2159-65
references.pmid: 2455217
references.citation: Nucleic Acids Res. 2000 Jan 1;28(1):235-42
references.pmid: 10592235
references.citation: FASEB J. 2002 Jul;16(9):1093-5
references.pmid: 12087071
references.citation: Mol Microbiol. 2005 Mar;55(5):1423-34
references.pmid: 15720551
references.citation: Blood. 1998 Sep 15;92(6):2147-56
references.pmid: 9731074
references.citation: PLoS One. 2007 Mar 28;2(3):e336
references.pmid: 17389925
references.citation: Protein Sci. 2006 Jun;15(6):1433-40
references.pmid: 16731977
references.citation: Am J Physiol Lung Cell Mol Physiol. 2010 Mar;298(3):L462-71
references.pmid: 20061440
references.citation: Nature. 2003 Aug 14;424(6950):783-8
references.pmid: 12917687
references.citation: Br J Haematol. 2003 Sep;122(6):1014-23
references.pmid: 12956774
references.citation: Transfus Med. 2002 Jun;12(3):205-11
references.pmid: 12164140
references.citation: Cancer Lett. 2008 Aug 28;267(2):226-44
references.pmid: 18579287
references.citation: Hum Genet. 1995 Apr;95(4):407-10
references.pmid: 7705836
references.citation: Br J Haematol. 2000 Feb;108(2):448-54
references.pmid: 10691880
references.citation: Proc Natl Acad Sci U S A. 1999 Aug 17;96(17):9459-64
references.pmid: 10449714
references.citation: Biochem Biophys Res Commun. 2007 May 11;356(3):816-21
references.pmid: 17382291
references.citation: Anal Chem. 2005 Dec 1;77(23):7547-55
references.pmid: 16316161
references.citation: Blood. 1995 Feb 1;85(3):622-6
references.pmid: 7833467
references.citation: J Exp Med. 1995 Apr 1;181(4):1311-7
references.pmid: 7699323
references.citation: Proc Natl Acad Sci U S A. 2010 Mar 30;107(13):5967-71
references.pmid: 20231434
references.citation: Vet Immunol Immunopathol. 2009 Mar 15;128(1-3):178-83
references.pmid: 19026455
references.citation: Arthritis Rheum. 2006 Jun;54(6):2022-6
references.pmid: 16732566
references.citation: Eur J Immunol. 2009 Feb;39(2):342-51
references.pmid: 19130487
references.citation: Methods Enzymol. 1990;185:60-89
references.pmid: 2199796
references.citation: Trends Parasitol. 2010 Jul;26(7):329-33
references.pmid: 20382562
references.citation: PLoS One. 2009 Dec 14;4(12):e8297
references.pmid: 20011536
references.citation: BMC Cancer. 2004 Jun 23;4:28
references.pmid: 15214968
references.citation: Proc Natl Acad Sci U S A. 2008 Jun 17;105(24):8363-8
references.pmid: 18523022
references.citation: Malar J. 2007 Dec 19;6:167
references.pmid: 18093292
references.citation: J Mol Biol. 1993 Jun 20;231(4):1049-67
references.pmid: 8515464
references.citation: Kidney Int. 2000 Oct;58(4):1546-56
references.pmid: 11012889
references.citation: J Immunol Methods. 1987 Sep 24;102(2):259-74
references.pmid: 2443575
references.citation: J Leukoc Biol. 1996 Jan;59(1):29-38
references.pmid: 8558064
references.citation: Trends Immunol. 2005 May;26(5):268-74
references.pmid: 15866240
references.citation: J Biol Chem. 1987 Feb 5;262(4):1596-601
references.pmid: 3805045
references.citation: Malar J. 2008 Oct 28;7:219
references.pmid: 18957099
references.citation: Immunol Lett. 2005 Jan 31;96(2):163-74
references.pmid: 15585320
references.citation: Nat Genet. 1995 Jun;10(2):224-8
references.pmid: 7663520
references.citation: Acta Biochim Pol. 2010;57(1):49-53
references.pmid: 20234884
references.citation: Cell Host Microbe. 2009 May 8;5(5):411-3; author reply 418-9
references.pmid: 19454340
references.citation: Trends Parasitol. 2007 May;23(5):193-6
references.pmid: 17360237
references.citation: Cytokine Growth Factor Rev. 2005 Dec;16(6):687-94
references.pmid: 16054417
references.citation: Cell Host Microbe. 2008 Jul 17;4(1):52-62
references.pmid: 18621010
references.citation: PLoS Med. 2007 Dec;4(12):e337
references.pmid: 18092885
references.citation: Methods Enzymol. 2009;461:191-206
references.pmid: 19480920
references.citation: Parasitol Res. 2010 Apr;106(5):1049-53
references.pmid: 20162434
references.citation: Protein Sci. 2009 Mar;18(3):619-28
references.pmid: 19241371
references.citation: Nucleic Acids Res. 1997 Dec 15;25(24):4876-82
references.pmid: 9396791
references.citation: J Mol Biol. 2001 Oct 26;313(3):473-8
references.pmid: 11676532
references.citation: Biochemistry. 1976 Mar 9;15(5):1137-44
references.pmid: 175830
references.citation: Proc Natl Acad Sci U S A. 1993 Nov 15;90(22):10793-7
references.pmid: 8248172
references.citation: Protein Expr Purif. 2005 May;41(1):207-34
references.pmid: 15915565
references.citation: J Biol Chem. 1995 Oct 27;270(43):25348-51
references.pmid: 7592697
references.citation: Science. 1993 Aug 27;261(5125):1182-4
references.pmid: 7689250
references.citation: Nature. 2006 Feb 9;439(7077):741-4
references.pmid: 16372020