Production of mouse monoclonal antibodies against human HLA-A3 for use in microlymphocytotoxicity technique

Abstract

Monoclonal antibody production using the hybridoma technology is one of the attractive areas of research. The aim of this study was to investigate the production of monoclonal antibodies to human HLA-A3 antigen for use in microlymphocytotoxicity technique. HLA-A3 antigen was purified from human leukocytes and injected to several mice. The spleen-derived cells of immunized animal were fused with P3-X63-Ag8 myeloma cells. Obtained hybridoma cells were grown in HAT medium and the supernatant of the cells were screened for antibodies using ELISA and microlymphocytotoxicity technique. Produced monoclonal antibody from the hybridoma clone, B1F10 showed the specificity for human HLA-A3. Furthermore, in this study, we encountered with clones of immortal hybrid cells with completely different features, they were adherent and non-producers for antibodies. This observation could imply the possible role of the antigen (HLA) and its nature in the expansion of cell types other than B cells in the spleen of mouse during immunization.