Preparation and Comparative Properties of Antisera against Glyco-3β, 12α-dihydroxy-5-cholen-24-oic Acid Linked to Bovine Serum Albumin at the C-3,C-15α, and C-24 Positions

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Anti glyco-3β, 12α-dihydroxy-5-cholen-24-oic acid antisera were prepared by immunizing rabbits with haptenbovine serum albumin (BSA) conjugates coupled at the C-3,C-15α, and C-24 positions on the bile acid molecule, and their properties were investigated by heterologous combination assay using ^<125>I-labeled tracer. The antiserum raised against the C-3 BSA conjugate showed poor titer and specificity, while the antisera from the other two conjugates showed satisfactorily high affinity constants (K_a = 5.0×10^8 and 7.0×10^8 M^<-1>) and reasonable specificity, exhibiting negligible cross-reactivities with other major human bile acids and cholesterol. Among the unsaturated bile acids tested, high reactivity was observed with tauro-3β, 12α-dihydroxy-5-cholen-24-oic acid, which suggested that bridge phenomena were significant in this assay system.
公益社団法人日本薬学会の論文
1989-08-25