Ozaki S, Tashiro A, Makino I, Nakagawa S, Yoshizawa I
J Lipid Res. 1979 Feb;20(2):240-5.
A sensitive and specific enzyme-linked immunoassay for the measurement of ursodeoxycholic acid in human serum was developed. Ursodeoxycholic acid conjugated to alkaline phosphatase (from calf intestine) was used as a tracer. An antiserum to ursodeoxycholic acid serum was raised in rabbits using ursodeoxycholic acid--bovine serum albumin conjugate as an antigen. The binding required 1 hr at 42 degrees C; separation of the bound tracer was achieved by addition of a second antibody, and alkaline phosphatase activity of this bound complex was measured colorimetrically. The ratio of bound to total enzyme activity decreased linearly with a logarithmic increase in ursodeoxycholic acid concentration from 20 to 900 pmol. The specificity and sensitivity of this enzyme-linked immunoassay were similar to those of a radioimmunoassay reported previously. The serum ursodeoxycholic acid levels measured by this method correlated well with those determined by gas--liquid chromatography and radioimmunoassay. Based on these findings, this enzyme-linked immunoassay of bile acid might be useful as a tool for the routine clinical analysis of serum bile acids.
开发了一种用于测定人血清中熊去氧胆酸的灵敏且特异的酶联免疫测定法。与碱性磷酸酶（来自小牛肠）偶联的熊去氧胆酸用作示踪剂。使用熊去氧胆酸-牛血清白蛋白偶联物作为抗原，在兔中制备了抗熊去氧胆酸血清。结合反应在42℃下需进行1小时；通过加入第二抗体实现结合示踪剂的分离，并通过比色法测定该结合复合物的碱性磷酸酶活性。随着熊去氧胆酸浓度从20至900 pmol呈对数增加，结合酶活性与总酶活性的比值呈线性下降。这种酶联免疫测定法的特异性和灵敏度与先前报道的放射免疫测定法相似。用该方法测定的血清熊去氧胆酸水平与气相色谱法和放射免疫测定法测定的结果相关性良好。基于这些发现，这种胆汁酸的酶联免疫测定法可能作为血清胆汁酸常规临床分析的一种工具。
