Glyco-3β-hydroxy-5-cholen-24-oic acid-bovine serum albumin (BSA) conjugate was prepared by coupling the hapten with the carrier protein through a 3-hemisuccinyl bridge.The antisera obtained from two rabbits elicited by this new antigen showed satisfactory affinity constants (Ka=1.16×107M-1and 2.28×107M-1) and reasonable specificity to glyco-3β-hydroxy-5-cholen-24-oic acid, exhibiting low cross-reactivities with cholesterol (0.01% and 0.27%) and the major human bile acids.Since the both antisera from two rabbits had somewhat different reactivities, with unsaturated bile acid they were able to be used for the determination.
