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Characterization of a new human monoclonal antibody directed against the Vel antigen

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
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Contributor : Laurent Jonchère Connect in order to contact the contributor
Submitted on : Wednesday, December 2, 2015 - 4:37:43 PM
Last modification on : Tuesday, October 19, 2021 - 5:56:21 PM

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Y. Danger, S. Danard, V. Gringoire, T. Peyrard, P. Riou, et al.. Characterization of a new human monoclonal antibody directed against the Vel antigen. Vox Sanguinis, Wiley, 2016, 110 (2), pp.172-178. ⟨10.1111/vox.12321⟩. ⟨hal-01237092⟩



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