Minder E I, Karlaganis G, Paumgartner G
J Lipid Res. 1979 Nov;20(8):986-93.
A radioimmunoassay for the determination of 3 beta-hydroxy-5-cholenoic acid in human serum has been developed, using 3 beta-hydroxy-5-cholenoyl-thyroglobulin as immunogen and 3 beta-hydroxy-5-cholenoylglycyl-125I histamine as radioactive ligand. The association constant was 6.3 X 10(8) l/mol. Cross reactivity with other bile acids of human serum was not detectable, but was 5.6% with cholesterol. Serum sample preparation included extraction of 3 beta-hydroxy-5-cholenoic acid from serum, solvolysis of sulfates, hydrolysis of conjugates, and separation from cholesterol by thin-layer chromatography. Serum concentrations of 3 beta-hydroxy-5-cholenoic acid were 0.23 +/- SD 0.12 mumol/l and 0.21 +/- SD 0.09 mumol/l in healthy males and females, respectively. In patients with primary biliary cirrhosis the serum concentration of 3 beta-hydroxy-5-cholenoic acid and the quotient 3 beta-hydroxy-5-cholenoic acid over total 3 alpha-hydroxy-bile acids (measured enzymatically) were significantly higher (P less than 0.02) than in patients with chronic active hepatitis or alcoholic cirrhosis. Analysis of 17 sera with elevated concentration of 3 beta-hydroxy-5-cholenoic acid by radioimmunoassay and capillary gas-liquid chromatography showed a close correlation (r = 0.91, slope = 0.97) between the results of the two methods.
已开发出一种用于测定人血清中3β-羟基-5-胆烯酸的放射免疫分析方法，使用3β-羟基-5-胆烯酰-甲状腺球蛋白作为免疫原，3β-羟基-5-胆烯酰甘氨酰-125I组胺作为放射性配体。缔合常数为6.3×10(8) l/mol。未检测到与人血清中其他胆汁酸的交叉反应，但与胆固醇的交叉反应为5.6%。血清样品制备包括从血清中提取3β-羟基-5-胆烯酸、硫酸盐的溶剂解、结合物的水解以及通过薄层色谱法与胆固醇分离。健康男性和女性血清中3β-羟基-5-胆烯酸的浓度分别为0.23±标准差0.12 μmol/l和0.21±标准差0.09 μmol/l。在原发性胆汁性肝硬化患者中，3β-羟基-5-胆烯酸的血清浓度以及3β-羟基-5-胆烯酸与总3α-羟基胆汁酸（酶法测定）的比值显著高于慢性活动性肝炎或酒精性肝硬化患者（P<0.02）。通过放射免疫分析和毛细管气液色谱法对17份3β-羟基-5-胆烯酸浓度升高的血清进行分析，结果显示两种方法的结果之间具有密切相关性（r = 0.91，斜率 = 0.97）。
