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Epitope mapping of the Dx5 antigen: an anti-COCO1 antibody defines a new type of autoantibody specific for the major intrinsic sperm protein.
The major sperm antigen was identified in 1992 as the Dx5 protein. Previously, it was shown that this antigen is implicated in the formation of the sperm membrane in rats and human. Nevertheless, the target epitope(s) has not yet been determined. For this reason, we investigated the epitope specificity of a monoclonal antibody (mAb) directed against a Dx5 derivative and developed by us (named “mAb Dx5.6D3”). The reactivity of this mAb was specifically directed against a conformational epitope of the Dx5 protein. In contrast with its reactivity on the sperm, this mAb reacted on the sperm membrane but not on the ciliary tract epithelium. Moreover, it did not react with the plasma membrane of any other kind of cells. This mAb was used for the mapping of the epitope of Dx5. Western blot experiments of proteins purified from Dx5-bearing sperm membranes demonstrated the existence of at least two distinct epitopes. We used a competitive immunoprecipitation assay to obtain evidence of an independent epitope region. The domain containing the antigen was defined by a