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Studies of a murine monoclonal antibody directed against DARC: reappraisal of its specificity.
Smolarek, Dorota; Hattab, Claude; Buczkowska, Anna; Kaczmarek, Radoslaw; Jarzab, Anna; Cochet, Sylvie; de Brevern, Alexandre G; Lukasiewicz, Jolanta; Jachymek, Wojciech; Niedziela, Tomasz; Grodecka, Magdalena; Wasniowska, Kazimiera; Colin Aronovicz, Yves; Bertrand, Olivier; Czerwinski, Marcin.
Afiliación
  • Smolarek D; Laboratory of Glycoconjugate Immunochemistry Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland; INSERM, UMR_S1134, F-75015 Paris, France; Institut National de la Transfusion Sanguine, F-75015 Paris, France; Division of Transplantation Immunology and Mucosal Biology,
  • Hattab C; INSERM, UMR_S1134, F-75015 Paris, France; Institut National de la Transfusion Sanguine, F-75015 Paris, France; Universite Paris-Diderot, Sorbonne Paris Cité, F-15013, Paris, France; Laboratory of Excellence GR-Ex, Paris, France.
  • Buczkowska A; Laboratory of Glycoconjugate Immunochemistry Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland; Institute of Biochemistry and Biophysics, Warsaw, Poland.
  • Kaczmarek R; Laboratory of Glycoconjugate Immunochemistry Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland.
  • Jarzab A; Laboratory of Medical Microbiology, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland.
  • Cochet S; INSERM, UMR_S1134, F-75015 Paris, France; Institut National de la Transfusion Sanguine, F-75015 Paris, France; Universite Paris-Diderot, Sorbonne Paris Cité, F-15013, Paris, France; Laboratory of Excellence GR-Ex, Paris, France.
  • de Brevern AG; INSERM, UMR_S1134, F-75015 Paris, France; Institut National de la Transfusion Sanguine, F-75015 Paris, France; Universite Paris-Diderot, Sorbonne Paris Cité, F-15013, Paris, France; Laboratory of Excellence GR-Ex, Paris, France.
  • Lukasiewicz J; Laboratory of Microbial Immunochemistry and Vaccines Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland.
  • Jachymek W; Laboratory of Microbial Immunochemistry and Vaccines Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland.
  • Niedziela T; Laboratory of Microbial Immunochemistry and Vaccines Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland.
  • Grodecka M; Laboratory of Glycoconjugate Immunochemistry Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland.
  • Wasniowska K; Laboratory of Glycoconjugate Immunochemistry Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland; Institute of Physiotherapy, Faculty of Physical Education and Physiotherapy, Opole University of Technology, Opole, Poland.
  • Colin Aronovicz Y; INSERM, UMR_S1134, F-75015 Paris, France; Institut National de la Transfusion Sanguine, F-75015 Paris, France; Universite Paris-Diderot, Sorbonne Paris Cité, F-15013, Paris, France; Laboratory of Excellence GR-Ex, Paris, France.
  • Bertrand O; INSERM, UMR_S1134, F-75015 Paris, France; Institut National de la Transfusion Sanguine, F-75015 Paris, France; Universite Paris-Diderot, Sorbonne Paris Cité, F-15013, Paris, France; Laboratory of Excellence GR-Ex, Paris, France.
  • Czerwinski M; Laboratory of Glycoconjugate Immunochemistry Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland; Institute of Physiotherapy, Faculty of Physical Education and Physiotherapy, Opole University of Technology, Opole, Poland.
PLoS One ; 10(2): e0116472, 2015.
Article en En | MEDLINE | ID: mdl-25706384
Duffy Antigen Receptor for Chemokines (DARC) plays multiple roles in human health as a blood group antigen, a receptor for chemokines and the only known receptor for Plasmodium vivax merozoites. It is the target of the murine anti-Fy6 monoclonal antibody 2C3 which binds to the first extracellular domain (ECD1), but exact nature of the recognized epitope was a subject of contradictory reports. Here, using a set of complex experiments which include expression of DARC with amino acid substitutions within the Fy6 epitope in E. coli and K562 cells, ELISA, surface plasmon resonance (SPR) and flow cytometry, we have resolved discrepancies between previously published reports and show that the basic epitope recognized by 2C3 antibody is 22FEDVW26, with 22F and 26W being the most important residues. In addition, we demonstrated that 30Y plays an auxiliary role in binding, particularly when the residue is sulfated. The STD-NMR studies performed using 2C3-derived Fab and synthetic peptide corroborated most of these results, and together with the molecular modelling suggested that 25V is not involved in direct interactions with the antibody, but determines folding of the epitope backbone.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Receptores de Superficie Celular / Sistema del Grupo Sanguíneo Duffy / Anticuerpos Monoclonales / Epítopos Límite: Animals / Humans Idioma: En Revista: PLoS One Asunto de la revista: CIENCIA / MEDICINA Año: 2015 Tipo del documento: Article Pais de publicación: Estados Unidos
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Receptores de Superficie Celular / Sistema del Grupo Sanguíneo Duffy / Anticuerpos Monoclonales / Epítopos Límite: Animals / Humans Idioma: En Revista: PLoS One Asunto de la revista: CIENCIA / MEDICINA Año: 2015 Tipo del documento: Article Pais de publicación: Estados Unidos