IMMUNOLOGICAL CHARACTERIZATION OF OVIDUCTAL GLYCOPROTEINS ASSOCIATED WITH THE ZONA-PELLUCIDA OF THE GOLDEN-HAMSTER EGG

Three monoclonal antibodies (MAbs) were produced against materials obtained from the golden hamster oviduct. The MAbs (C11E8, C8B11, and A3D5) selectively reacted with the zona pellucida (ZP) of oviductal eggs. An immunohistochemical study demonstrated that all MAbs bound strongly to the epithelial cells of the oviduct and, to a lesser extent, of the uterus, the cervix, and the vagina. Weak reactions were also observed with some other tissues. All immunohistochemical reactions were completely eliminated by treatment of tissues with periodic acid, suggesting that the antigenic determinants that react with the MAbs are carbohydrate in nature. Antigens that reacted with the MAbs were characterized by immunoblotting analyses of 1- or 2-dimensional polyacrylamide gels after electrophoresis to fractionate tissue extracts under reducing conditions. With the oviductal extract, C11E8 and A3D5 specifically bound to broad bands that corresponded to macromolecules of more than 200 and 160 kD, respectively, whereas C8B11 reacted with a broad range of macromolecules, with the strongest reactivity being detected at molecular weights (MWs) higher than 160 kD. The macromolecules that reacted with the MAbs had carbamilation trains, suggestive of extensive microheterogeneity with respect to charge and size. Similar substances of high MW were not detected in extracts of tissues from the other reproductive organs. In addition, one MAb-positive oviductal antigen bound to the ZP of ovarian eggs, but the others did not bind to the ZP of ovarian eggs. This result indicates that the binding activity is specific for the oviductal glycoproteins. We have thus established clones that produce 3 different MAbs with strong affinity for oviduct-specific glycoproteins, one of which is associated with the ZP of the golden hamster.