[Production of monoclonal antibodies specific for the ABO blood group and rhesus D antigens]

We report, in this work, the techniques to obtain four monoclonal antibodies specific of erythrocytes antigens. Three of this antibodies, react with the ABO Blood Groupe System (A,B and AB), are produced by three mouse hybridomas (M18-2F11, M18-4F6 et M19-45D6), obtained by fusion of Sp2/0 mouse myeloma cell with spleen cell of balb/c mice immunized with human red blood cells and selected on selectif medium (Hypoxanthin, Aminoptérin, Thymidin) (HAT) and cloned by four limiting dilutions. Where as the fourth, it is an human monoclonal antibodies against Rhesus (D) produced by heterohybridomas, realized by fusion of X63 mouse myeloma cell with human B lymphocytes, from actively immunized persons by D antigen, that are purified and transformed by Epstein-Barr virus (EBV), and selected on selectif medium (HAT), presence of ouabain and then cloned by four limiting dilutions. The specificity of the antibodies produced, has been determined by direct hemagglutinin for the mouse monoclonal antibodies and artificial for the human anti-D. The determination of isotype the heavy and light chains is released by the technique immunoenzymatique (ELISA) and immunofixation has shown that mouse monoclonal antibodies belong to class IgM kappa, and human monoclonal antibodies anti-D belong to class IgG lambda.