Johnson P K, Messenger L J
Clin Chem. 1986 Feb;32(2):378-81.
In this immunoassay for disopyramide in serum, to form the label, disopyramide is covalently attached to a fluorogenic enzyme substrate, 7-beta-galactosylcoumarin-3-carboxylic acid, which is nonfluorescent under the conditions of the assay. Hydrolysis, catalyzed by beta-galactosidase, yields a fluorescent product. As a result of competitive protein binding reactions between drug and label for limited antibody binding sites, this fluorescence is proportional to disopyramide concentration. The assay is sensitive to less than 0.5 mg of disopyramide per liter. Results obtained with either the semi-automated procedure (Ames TDA) or the fully automated (Optimate) procedure correlated well with those obtained by liquid chromatography (r = 0.98 and 0.99). For commercial controls containing various concentrations of the drug, the respective coefficients of correlation were 1.00 and 0.99 for the TDA and Optimate procedures. Within-run CVs for the two procedures were less than or equal to 5.1%, overall CVs less than or equal to 6.5%.
在这种血清中丙吡胺免疫测定法中，为形成标记物，丙吡胺与一种荧光酶底物7-β-半乳糖基香豆素-3-羧酸共价连接，该底物在测定条件下无荧光。在β-半乳糖苷酶催化下发生水解，产生荧光产物。由于药物与标记物之间针对有限抗体结合位点的竞争性蛋白质结合反应，这种荧光与丙吡胺浓度成正比。该测定法对每升血清中低于0.5毫克的丙吡胺敏感。通过半自动程序（Ames TDA）或全自动程序（Optimate）获得的结果与通过液相色谱法获得的结果相关性良好（r = 0.98和0.99）。对于含有不同浓度药物的商业对照品，TDA和Optimate程序各自的相关系数分别为1.00和0.99。两种程序的批内变异系数小于或等于5.1%，总变异系数小于或等于6.5%。
