title: Characterization of a new human monoclonal antibody directed against the Vel antigen.
abstract: OBJECTIVE
  The Vel blood group antigen is a poorly characterized high-prevalence antigen. Until now, anti-Vel antibodies have been observed in only alloimmunized Vel-negative individuals. In this study, we aimed to establish a human hybridoma cell line secreting the first anti-Vel monoclonal antibody (mAb), clone SpG213Dc.
  
  
  METHODS
  Peripheral blood lymphocytes from a French Vel-negative woman with anti-Vel in her plasma were transformed with Epstein-Barr virus and then hybridized with the myeloma cell line Sp2/O-Ag14 using the polyethylene glycol (PEG) method. A specific anti-Vel mAb was successfully produced and was extensively characterized by serological, flow cytometry and Western blot analyses.
  
  
  RESULTS
  One human anti-Vel-secreting clone was produced and the secreted anti-Vel mAb (SpG213Dc) was examined. The specificity of the SpG213Dc mAb was assessed by its reactivity against a panel of nine genotyped RBCs including, respectively, three Vel-negative and six Vel-positive (three wild-type homozygous and three heterozygous) samples using flow cytometry method. Vel-positive RBCs were specifically stained and were subsequently used to perform Western blot and immunoprecipitation analysis of the Vel antigen.
  
  
  CONCLUSIONS
  Serological characterization of the new monoclonal anti-Vel SpG213Dc showed a heterogeneous level of expression of the Vel antigen on the different RBCs. Our results suggest that the mAb SpG213Dc can be reliably used as a blood grouping reagent, thus allowing the mass-scale phenotyping of blood donors to strengthen rare blood banks with Vel-negative RBC units.
authors.affiliation: EFS Bretagne, Rennes, France.|Structure Fédérative BioSit UMS 3480 CNRS-US18 Inserm, Rennes, France.
authors.forename: Y
authors.initials: Y
authors.lastname: Danger
authors.affiliation: EFS Bretagne, Rennes, France.
authors.forename: S
authors.initials: S
authors.lastname: Danard
authors.affiliation: EFS Bretagne, Rennes, France.
authors.forename: V
authors.initials: V
authors.lastname: Gringoire
authors.affiliation: Institut National de la Transfusion Sanguine, Département Centre National de Référence pour les Groupes Sanguins, Paris, France.|INSERM UMR_S1134, Paris, France.|Laboratoire d'Excellence LABEX GR-Ex, Paris, France.
authors.forename: T
authors.initials: T
authors.lastname: Peyrard
authors.affiliation: EFS Bretagne, Rennes, France.
authors.forename: P
authors.initials: P
authors.lastname: Riou
authors.affiliation: EFS Bretagne, Rennes, France.
authors.forename: G
authors.initials: G
authors.lastname: Semana
authors.affiliation: EFS Bretagne, Rennes, France.|Structure Fédérative BioSit UMS 3480 CNRS-US18 Inserm, Rennes, France.
authors.forename: F
authors.initials: F
authors.lastname: Vérité
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D001789:Blood Group Antigens / Q000276:immunology*; D005260:Female; D006801:Humans; D008565:Membrane Proteins / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D001789:Blood Group Antigens; D008565:Membrane Proteins; C584052:SMIM1 protein, human
keywords: SMIM1; Vel blood group antigen; human monoclonal antibody