T₃-BSA hapten conjugates were prepared by controlling pH and the molar ratio of BSA to T₃, and identified spectrophotometically. EDC, the coupling agent of BSA to T₃, is converted to water soluble urea derivertives for coupling reaction. In order to improve the conjugation efficiency of T₃-BSA, therefore, 9M urea was applied to the reaction mixture.
The molar ratios of T₃to BSA in the prepared conjugates were 2.0-4.7 in the abscence of urea. The values are proportional to the increasing ratios of T₃to BSA in reaction media. However, the ratios in urea media were decreased to 1.0-3.9, due to interfering participation of the precipitates formed in the reaction course. The results indicate that urea is not effective on the preparation of T₃-BSA. For an efficient preparation of T₃-BSA, the condition under the high ratio of T3 to BSA should be established as far as possible to minimize the formation of precipitate during the reaction time.
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