Monoclonal antibodies to chick crystallins

Several monoclonal antibodies against chick crystallins were obtained. Immunoblot analysis indicated that the monoclonal antibodies distinguish not only the three classes of crystallin (alpha, beta and delta), but also their subclasses. Monoclonal antibodies alpha 2 and alpha 1 reacted exclusively with alpha A and alpha B, respectively, demonstrating that alpha-crystallin subclasses of the chicken are antigenically distinct. An immunohistological study utilizing these monoclonal antibodies showed that the two alpha-crystallin subclasses, alpha A and alpha B, are co-expressed in the same cells and are more concentrated in the epithelium than the fibers in 14-day-old chick embryo lenses. However, immunofluorescence suggested different distribution of alpha-crystallin subclasses within an epithelial cell. alpha A is distributed evenly in the cytoplasm, but alpha B is more concentrated in the fiber-proximal side. Using anti-beta monoclonal antibodies, it was shown that beta-crystallins are divided into three distinct subclasses according to their antigenicity: 27-kDa and 25-kDa beta-crystallins are recognized by beta 1 antibody, 26-kDa and 19-kDa beta-crystallins by beta 2 antibody, and 35-kDa beta-crystallins by beta 3 antibody. All of the anti-delta-crystallin monoclonal antibodies (delta 1 to delta 3) obtained here bound to all delta-crystallin molecular species separable by isoelectrofocusing.