The preparation and antigenic properties of estradiol 3-sulfate 17-glucuro-nide-bovine serum albumin (BSA) conjugate in which the hapten is linked to the carrier protein through an (O-carboxymethyl)oxime bridge at the C-6 position on the steroid nucleus, have been described. Coupling of 6-oxoestradiol (O-carboxymethyl)oxime 3-sulfate 17-glucuronide acetate-methyl ester with BSA by the activated ester method followed by removal of the protecting groups with alkali provided the desired conjugate. The antisera raised against the conjugate were highly specific to the double conjugate, estradiol 3-sulfate 17-glucuronide, discriminating from unconjugated and ring A or D monoconjugated estrogens. The specificity of antisera elicited has been discussed on the basis of stereochemistry.
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