Characterization of a new human monoclonal antibody directed against the Vel antigen. Issue 2 (18th September 2015)

• Record Type: Journal Article
• Authors: Danger, Y.; Danard, S.; Gringoire, V.; Peyrard, T.; Riou, P.; Semana, G.; Vérité, F.
• Is Part Of: Vox sanguinis. Volume 110:Issue 2(2016)
• Page Start: 172
• Page End: 178
• Publication Date: 2015-09-18
• Subjects: human monoclonal antibody -- SMIM1 -- Vel blood group antigen; Blood -- Periodicals; Blood -- Transfusion -- Periodicals; Immunohematology -- Periodicals; Immunopathology -- Periodicals; 615.39
• DOI: 10.1111/vox.12321
• Languages: English
• ISSNs: 0042-9007

Abstract

Abstract : Background and Objectives: 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. Materials and 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. Conclusion: 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.