title: Simultaneous detection of two different cell surface antigens by electron microscopy by means of multivalent hybrid antibody with double specificity.
abstract: Multivalent hybrid antibody (MHA) complexes with dual specificity were prepared by combining two antibodies of different specificities, one against ferritin (Fer), the other against horseradish peroxidase (HRP), with protein A of Staphylococcus aureus (SpA). Electron microscopy of mouse spleen lymphocytes and thymocytes (previously coated with mouse IgG anti-Thy-1 antibody) treated with IgG anti-Fer/SpA/IgG anti-mouse Ig complex and Fer gave better resolution and higher accuracy than previously obtained with IgG anti-HRP/SpA/IgG anti-mouse Ig complex and HRP (Mandache et al., 1980). Surface Thy-1 alloantigen and Fc receptor (charged with human IgG) treated with a mixture of IgG anti-Fer/SpA/IgG anti-Thy-1 and IgG anti-HRP/SpA/IgG anti-human Fab could be simultaneously detected on the thymocyte surface by either light or electron microscopy using Fer and HRP. The concomitant visualization of Thy-1 alloantigen (with Fer) and FcR (with HRP) on mouse thymocyte clearly shows that their distribution is largely independent and that the amount of Thy-1 antigen is greater. These results show that electron microscopy with a mixture of MHA is a useful technique for simultaneous location of two antigenic markers on the cell surface.
mesh_terms: D000818:Animals; D000918:Antibody Specificity; D000954:Antigens, Surface; D002645:Chickens; D005293:Ferritins / Q000276:immunology; D006735:Horseradish Peroxidase / Q000276:immunology; D006736:Horses; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D008214:Lymphocytes / Q000648:ultrastructure*; D051379:Mice; D008808:Mice, Inbred CBA; D011817:Rabbits; D012397:Rosette Formation; D012756:Sheep; D013205:Staphylococcal Protein A / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology* / Q000648:ultrastructure; D013997:Time Factors
chemical_list: D000954:Antigens, Surface; D007140:Immunoglobulin Fab Fragments; D013205:Staphylococcal Protein A; D005293:Ferritins; D006735:Horseradish Peroxidase