Abstract
Monoclonal antibodies, raised against partially purified AMH, have allowed the purification of the hormone to homogeneity, and the development of radioimmunological and immunocytochemical methods, which could however be applied only to bovine AMH (bAMH), because of the zoospecificity of the first generation of monoclonal antibodies. Wishing to use these methods in clinical studies, we have sought to obtain antibodies recognizing human AMH. Two methods have been used. A second generation of monoclonal antibodies was obtained by immunization with pure bAMH. Thirteen clones, screened through an immunodot procedure using bAMH as antigen, have been identified. Their zoospecificity has been tested by using them as first antibody in an immunocytochemical reaction performed on testicular sections of different species. Caprine and ovine AMH are recognized by all monoclonal antibodies reacting with bAMH, most of these are IgG clones with high affinity. Three IgM clones recognize pig AMH, and one binds to human AMH. In parallel, a polyclonal antibody has been raised in a rabbit immunized with pure bovine AMH. This antibody is not zoospecific, and has allowed an immunocytochemical study of AMH in human testicular tissue, through an avidin-biotin-immunoperoxidase method, performed upon cryostat sections of testicular biopsies or autopsy material. AMH has been identified in the Sertoli cells of 6 human fetuses, aged 16 to 22 weeks, and 4 boys aged 1 month to 5 years. This method can be used to study testicular AMH in intersex disorders.
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Josso, N., Picard, J., Legeai, L. et al. Polyclonal versus monoclonal antibodies: application to the study of human anti-Müllerian hormone. Pediatr Res 18, 1211 (1984). https://doi.org/10.1203/00006450-198411000-00061
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DOI: https://doi.org/10.1203/00006450-198411000-00061