Wilson R
Research Centre for Advanced Science and Technology, University of Tokyo, Japan.
Analyst. 1992 Oct;117(10):1547-51. doi: 10.1039/an9921701547.
Thyroid stimulating hormone (TSH) regulates the function of the thyroid gland. Its determination at low concentrations in serum is useful in the diagnosis of hyperthyroidism. In this paper, it is detected using a spectrophotometric enzyme-amplified immunoassay. The reporter enzyme is alkaline phosphatase and its substrate is flavin adenine dinucleotide phosphate (FADP). Reaction with alkaline phosphatase converts FADP into flavin adenine dinucleotide (FAD), which, unlike FADP, re-activates apo-D-amino acid oxidase (apo-AOD). Re-activation of apo-AOD allows the product of the reporter enzyme to be amplified. The lower limit of detection for TSH by this method is 0.06 microU cm-3. This compares with 0.54 microU cm-3 for an identical assay in which p-nitrophenyl phosphate was the substrate for alkaline phosphatase. Contaminating alkaline phosphatase was removed from the reagents by affinity chromatography.
促甲状腺激素（TSH）调节甲状腺的功能。测定血清中低浓度的TSH有助于诊断甲状腺功能亢进症。在本文中，采用分光光度酶放大免疫测定法对其进行检测。报告酶为碱性磷酸酶，其底物为黄素腺嘌呤二核苷酸磷酸（FADP）。与碱性磷酸酶反应将FADP转化为黄素腺嘌呤二核苷酸（FAD），与FADP不同，FAD可重新激活脱辅基-D-氨基酸氧化酶（apo-AOD）。apo-AOD的重新激活使报告酶的产物得以放大。该方法检测TSH的下限为0.06微单位/立方厘米。相比之下，以对硝基苯磷酸作为碱性磷酸酶底物的相同检测方法的下限为0.54微单位/立方厘米。通过亲和层析从试剂中去除了污染的碱性磷酸酶。
