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Sensitive fluorometric detection of alkaline phosphatase using a water-soluble conjugated polymer

机译：使用水溶性共轭聚合物的灵敏荧光检测碱性磷酸酶

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In this paper, we developed a new fluorescence method for highly sensitive and selective detection of the alkaline phosphatase (ALP) activity with phenyl phosphate (PP) as the enzyme substrate and water-soluble conjugated polymer PPESO3 as a fluorescent probe. Phenyl phosphate could be hydrolyzed to phenol via the ALP-catalyzed reaction. In the presence of horse radish peroxidase (HRP) and H2O2, phenol was rapidly oxidized to quinone which could efficiently quench the fluorescence of the conjugated polymer PPESO3. The fluorescence intensity change of PPESO3 is proportional to the concentration of ALP. Thus, we established a fluorescence method for ALP activity detection based on a water-soluble fluorescent conjugated polymer-enzyme hybrid system. Under the optimized conditions, a linear correlation was established between the fluorescence intensity ratio I/I-0 (I-0 and I were the fluorescence intensity of the sensing system in the absence and presence of ALP, respectively) and the concentration of ALP in the range of 0-30 U L-1, and the detection limit for ALP was 0.5 U L-1. The present method was applied to the determination of ALP in spiked human serum samples with satisfactory results.
机译：在本文中，我们开发了一种新的荧光方法，用于以磷酸苯酯（PP）为酶底物和水溶性共轭聚合物PPESO3作为荧光探针的高灵敏度和选择性检测碱性磷酸酶（ALP）活性。磷酸苯酯可通过ALP催化反应水解为苯酚。在辣根过氧化物酶（HRP）和H2O2的存在下，苯酚被快速氧化为醌，可以有效地淬灭共轭聚合物PPESO3的荧光。 PPESO3的荧光强度变化与ALP浓度成正比。因此，我们建立了一种基于水溶性荧光共轭聚合物-酶杂化体系的ALP活性检测的荧光方法。在最佳条件下，荧光强度比I / I-0（I-0和I分别是不存在和存在ALP时传感系统的荧光强度）与ALP浓度之间的线性相关关系范围为0-30 U L-1，ALP的检出限为0.5 U L-1。本方法用于加标人血清样品中ALP的测定，结果令人满意。

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