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一种用于血清血管紧张素转换酶的灵敏荧光测定法。

A sensitive fluorimetric assay for serum angiotensin-converting enzyme.

作者信息

Friedland J, Silverstein E

出版信息

Am J Clin Pathol. 1976 Aug;66(2):416-24. doi: 10.1093/ajcp/66.2.416.

Abstract

A simple, rapid, highly sensitive and reproducible assay for angiotensin-converting enzyme in untreated serum is described. It is based on the conversion of the substrate analog, hippuryl-L-histidyl-L-leucine (5 mM in 0.1 M K phosphate, pH 8.3-0.3 M NaCl) to hippurate and L-histidyl-L-leucine, which is quantified spectrofluorimetrically (lamdba excitation = 360 nm; lamdba fluorescence = 500 nm) by formation of a fluorescent adduct with ophthaldialdehyde. The chloride requirement and inhibition and activation patterns correspond to those for angiotensin-converting enzyme. The Km for hippuryl-L-histidyl-L-leucine was 1.33 mM. The mean value of serum angiotensin-converting enzyme for 58 normal human subjects (mean age, 32 years; range 19-57) was 32.2 +/- 1.30 (SE), with a standard deviation of 9.87 nmol/min/ml serum. The assay is useful for the diagnosis and possible management of sarcoidosis and may have other applications in the future.

摘要

本文描述了一种用于检测未处理血清中血管紧张素转换酶的简单、快速、高度灵敏且可重复的检测方法。该方法基于底物类似物马尿酸-L-组氨酸-L-亮氨酸(在0.1M磷酸钾,pH8.3-0.3M氯化钠中为5mM)转化为马尿酸和L-组氨酸-L-亮氨酸,通过与邻苯二甲醛形成荧光加合物,采用荧光分光光度法(激发波长=360nm;荧光波长=500nm)对其进行定量。对氯化物的需求以及抑制和激活模式与血管紧张素转换酶的情况相符。马尿酸-L-组氨酸-L-亮氨酸的米氏常数为1.33mM。58名正常人类受试者(平均年龄32岁;范围19-57岁)血清血管紧张素转换酶的平均值为32.2±1.30(标准误),标准差为9.87nmol/分钟/毫升血清。该检测方法对结节病的诊断和可能的治疗有用,未来可能还有其他应用。

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