Study of H-Y antigen in abnormal sex determination with monoclonal antibody and an ELISA

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Abstract
A newly developed enzyme‐linked immunosorbent assay (ELISA) has been applied to the study of H‐Y antigen in cases of XY, XYY, and X,dic Y gonadal dysgenesis, testicular feminization syndrome, XXXXY syndrome, and XX true hermaphroditism. Monoclonal H‐Y antibody was absobed with cells from each of eight patients and from normal male and female controls, and then reacted with a plated antigen source in a system subsuming the addition of biotinylated secondary antibody, avidin‐biotin‐enzyme complex and substrate, and thereby the generation of a color. Positive absorption decreased the reaction, and this allowed sensitive measurement of H‐Y phenotype in an electronic optical density reader. The ELISA obviates many of the technical difficulties encountered in complement‐mediated cytotoxicity systems and can be used in the study of clinical cases of aberrant sex determination and in the evaluation of current models of the genetics of sex determination.