Estriol antisera were raised and studied for the use in radioimmunoassay (RIA) of urinary estrogen as an aid of diagnosis of feto-placental function. Antisera to estriol-related six haptens, i.
e
., estriol 16-glucuronide (
E3
-16-G), estriol 16-hemisuccinate (
E3
-16-succ), estriol 17-hemisuccinate (
E3
-17-succ), estriol 16, 17-dihemisuccinate (
E3
-16, 17-succ), estriol
3
-carboxymethylether (
E3
-
3
-CME) and estriol
3
-glucuronide (
E3
-
3
-G), were obtained by immunizing rabbits with the respective haptenbovine serum albumin (BSA) conjugate. Antiserum to
E3
-16-G-BSA reacted significantly with
E3
-16-G, C-17 conjugated estrogens and free estrogens. But it did not react with C-
3
conjugated estrogens and other steroids. Similar properties were observed with antisera to
E3
-16-succ-BSA,
E3
-17-succ-BSA and
E3
-16, 17-succ-BSA, but their reactivities with C-16 or C-17 conjugated estrogens were slightly lower than that of anti-
E3
-16-G-BSA serum. Among antisera which were raised against estriol linked to BSA via hemisuccinate varying in linking posirion, anti-
E3
-16-succ-BSA serum highly reacted with
E3
-16-G, while anti-
E3
-17-succ-BSA did with
E3
-17-G. Anti-
E3
-16, 17-succ-BSA serum reacted with C-16 and C-17 conjugated estrogens to nearly the same extent. Antisera to both
E3
-
3
-CME-BSA and
E3
-
3
-G-BSA reacted with estrogens conjugated at C-
3
position, but failed to react with C-16 or C-17 conjugated estrogens. The latter antiserum reacted more specifically with
E3
and
E3
3
-G than the former. Estrogen values in pregnancy urine samples estimated by the RIA by use of anti-
E3
-16-G serum were not influenced by other components in urine, and showed a good correlation of y=0.81x-1.28, r=0.983 with those determined by the currently available colorimetric method with
E3
-kit.
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