| Abstract: | Multivalent hybrid antibody (MHA) complexes with double specificity were prepared by combining two different antibodies, one specific against ferritin (Fer), the other against horseradish peroxidase using protein A from Staphylococcus aureus (SpA). Electron microscopy of mouse spleen lymphocytes and thymocytes (the latter coated with mouse anti-Thy-1 antibody) reacted with anti-HRP/SpA/anti-mIg complex and HRP showed the pattern of surface Ig receptors visualized by the specific peroxidase labelling. When IgG anti-Fer/SpA/IgG-anti-mIg complex was applied to the same cellular stuff, better resolution and higher accuracy were obtained although the distribution was similar. Surface Thy-1 alloantigen and Fe receptors (charged with hIgG) were simultaneously detected on the thymocyte surface by using in the same way a mixture of anti-Fer/SpA/anti-Thy-1 and anti-HRP/SpA/anti-Fab. The concomitant ultrastructural visualization of Thy-1 (with Fer) and Fc (with HRP) on mouse thymocytes clearly showed that their distribution was mainly independent and that the amount of Thy-1 antigen prevailed. These data show that electron microscopy with a mixture of MHA may be a useful technique for the concomitant location of two antigenic determinants on the cell surface. |
