Rimet O, Chauvet M, Bourdeaux M, Briand C
Laboratoire de Physique Pharmaceutique, Faculté de Pharmacie, Marseille, France.
J Biochem Biophys Methods. 1987 Sep;14(6):335-42. doi: 10.1016/0165-022x(87)90027-3.
In an effort to study the level of dihydrofolate reductase (DHFR), the main molecular target of antifolate drugs, in healthy and malignant tissues of human origin, a new and convenient fluorometric enzymatic assay has been developed. The technique measures the overall decrease in fluorescence emission at 454 nm (lambda ex = 342 nm) due to the contributions from coenzyme oxidation and substrate reduction. This technique was developed by using an enzyme purified from beef liver. All criteria of quality were checked: sensitivity, reproducibility and specificity made it suitable for low activity measurements. It was successfully applied to human tissue crude extracts.
为了研究抗叶酸药物的主要分子靶点二氢叶酸还原酶（DHFR）在人源健康组织和恶性组织中的水平，已开发出一种新的便捷荧光酶法测定方法。该技术通过测量由于辅酶氧化和底物还原导致的454nm处荧光发射的总体下降（激发波长λex = 342nm）来进行检测。此技术是利用从牛肝中纯化的一种酶开发的。检查了所有质量标准：灵敏度、重现性和特异性使其适用于低活性测量。该方法已成功应用于人体组织粗提物的检测。
