Immunogen was testosterone-3-carboxymethyl oxime-bovine serum albumin(To-3-CMO-BSA)

and tracer ligand was testosterone-3-carboxymethyl oxime-4-(10-methy)-acridinium-9-carboxylate(To-3-CMO-AEPMAC). A chemiluminescene immunoassay of testosterone(To-CLIA) was established

on the equilibrium method

by raising the antiserum in rabbits against To-3-CMO-BSA. The data was processed by the four-parameters logistic function program. The intra- and inter-assay CV of To-CLIA were 5.78% and 3.30%

respectively

the recovery rate

107.28%. The antiserum provided a linear response from 19.3 to 264.6ng/L with ED 50 of 82.15ng/L and correlation coefficient of curve fitting

0.9967. The cross reactivity with estrone

estradiol

estriol and progesterone were less 1.35%. Non-specific bound rate was 9.64%. The highest affinity constant could be obtained by "bridge effect" analysis with same structure of the "bridge" in the same position of testosterone.