使用自指示底物2,6-二氯-4-乙酰基苯磷酸酯的前列腺酸性磷酸酶测定法。

Prostatic acid phosphatase assay with self-indicating substrate 2,6-dichloro-4-acetylphenyl phosphate.

作者信息

Osawa S, Iida S, Yonemitsu H, Kuroiwa K, Katayama K, Nagasawa T

机构信息

Department of Laboratory Medicine, Chiba University Hospital, Japan.

出版信息

Clin Chem. 1995 Feb;41(2):200-3.

PMID:7874772

Abstract

We characterized six self-indicating substrates, synthesized as the derivative compounds of acetylphenyl phosphate, for serum prostatic acid phosphatase (PAP) activity. One of the substrates, 2,6-dichloro-4-acetylphenyl phosphate (DCAPP), is superior to others in terms of stability, affinity, and low Km for PAP. The hydrolyzed product, 2,6-dichloro-4-acetylphenol (DCAP), has a maximum absorption at 334.2 nm, a pKa of 4.15, and a molar absorptivity at 340 nm of 21,490 L.mol-1.cm-1 in citrate-HCl buffer, pH 5.4. PAP activity was assessed by subtracting tartaric acid-inhibited acid phosphatase activity from total acid phosphatase activity. Our assay system involving DCAPP is a unique kinetic method that shows good reproducibility, wide analytical dynamic range, and high specificity for PAP. Moreover, it is easily adaptable to automated analyzers because the product, DCAP, can be monitored at 340 nm.

摘要

我们对六种作为磷酸乙酰苯酯衍生化合物合成的自指示底物进行了表征，以检测血清前列腺酸性磷酸酶（PAP）的活性。其中一种底物2,6 - 二氯 - 4 - 乙酰基苯磷酸酯（DCAPP）在稳定性、亲和力以及对PAP的低米氏常数方面优于其他底物。水解产物2,6 - 二氯 - 4 - 乙酰基苯酚（DCAP）在334.2 nm处有最大吸收峰，pKa为4.15，在pH 5.4的柠檬酸盐 - 盐酸缓冲液中，340 nm处的摩尔吸光系数为21,490 L·mol⁻¹·cm⁻¹。通过从总酸性磷酸酶活性中减去酒石酸抑制的酸性磷酸酶活性来评估PAP活性。我们涉及DCAPP的检测系统是一种独特的动力学方法，具有良好的重现性、宽分析动态范围以及对PAP的高特异性。此外，它很容易适应自动分析仪，因为产物DCAP可以在340 nm处进行监测。