Mandache E, Mota G, Moldoveanu E, Moraru I, Ghetie V
J Immunol Methods. 1981;42(3):355-65. doi: 10.1016/0022-1759(81)90164-2.
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.
通过将两种具有不同特异性的抗体（一种抗铁蛋白（Fer），另一种抗辣根过氧化物酶（HRP））与金黄色葡萄球菌的蛋白A（SpA）相结合，制备了具有双重特异性的多价杂交抗体（MHA）复合物。用IgG抗Fer/SpA/IgG抗小鼠Ig复合物和Fer处理的小鼠脾淋巴细胞和胸腺细胞（先前用小鼠IgG抗Thy-1抗体包被）的电子显微镜观察，比先前用IgG抗HRP/SpA/IgG抗小鼠Ig复合物和HRP观察得到了更好的分辨率和更高的准确性（曼达切等人，1980年）。用IgG抗Fer/SpA/IgG抗Thy-1和IgG抗HRP/SpA/IgG抗人Fab的混合物处理的表面Thy-1同种异体抗原和Fc受体（负载人IgG），可以通过使用Fer和HRP的光学显微镜或电子显微镜在胸腺细胞表面同时检测到。小鼠胸腺细胞上Thy-1同种异体抗原（用Fer）和FcR（用HRP）的同时可视化清楚地表明它们的分布在很大程度上是独立的，并且Thy-1抗原的量更大。这些结果表明，用MHA混合物进行电子显微镜检查是一种用于在细胞表面同时定位两种抗原标记的有用技术。
