AbstractThe authors describe a sensitive fluorometric method for the determination of the activity of alkaline phosphatase (ALP). It is based on the use of a composite prepared consisting of flower-like cobalt oxyhydroxide (CoOOH) and copper nanoclusters (CuNCs). On formation of the CuNC-CoOOH aggregates, the fluorescence of the CuNCs is quenched by the CoOOH sheets. If, however, the CoOOH sheets are reduced to Co(II) ions in the presence of ascorbic acid (AA), fluorescence recovers. AA is formed in-situ by hydrolysis of the substrate ascorbic acid 2-phosphate (AA2P) as catalyzed by ALP. Thus, the ALP activity can be detected indirectly by kinetic monitoring of the increase in fluorescence, best at excitation/emission wavelengths of 335/410 nm. The assay allows ALP to be determined in 0.5 to 150 mU·mL−1 activity range and with a 0.1 mU·mL−1 detection limit. The method was successfully applied to the determination of ALP activity in (spiked) human serum samples. The assay has attractive features in being of the off-on type and immune against false positive results. Graphical AbstractA fluorescent bioassay is reported for the determination of the activity of alkaline phosphatase (ALP). It is exploiting the ascorbic acid (AA)-induced decomposition of nanoclusters composed of flower-like cobalt oxyhydroxide and copper nanoclusters. ALP catalyzes hydrolysis of ascorbic acid 2-phosphate (AA2P) and dephosphorylation to form AA.

中文翻译：

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花状羟基氧化钴和铜纳米团簇组成的纳米团簇作为荧光探针测定碱性磷酸酶的活性

摘要作者描述了一种用于测定碱性磷酸酶 (ALP) 活性的灵敏荧光法。它基于使用由花状羟基氧化钴 (CoOOH) 和铜纳米团簇 (CuNCs) 组成的复合材料。在形成 CuNC-CoOOH 聚集体时，CuNCs 的荧光被 CoOOH 片淬灭。然而，如果 CoOOH 片在抗坏血酸 (AA) 存在下被还原为 Co(II) 离子，则荧光会恢复。AA 是在 ALP 催化下通过底物抗坏血酸 2-磷酸 (AA2P) 的水解原位形成的。因此，ALP 活性可以通过荧光增加的动力学监测间接检测，最好在 335/410 nm 的激发/发射波长。该测定允许在 0.5 到 150 mU·mL-1 的活性范围内测定 ALP，并以 0. 1 mU·mL−1 检测限。该方法已成功应用于测定（加标）人血清样品中的 ALP 活性。该测定具有吸引人的特点，即关闭型和对假阳性结果免疫。图形摘要报道了一种荧光生物测定法，用于测定碱性磷酸酶 (ALP) 的活性。它正在利用抗坏血酸 (AA) 诱导的由花状羟基氧化钴和铜纳米团簇组成的纳米团簇分解。ALP 催化抗坏血酸 2-磷酸 (AA2P) 的水解和去磷酸化以形成 AA。图形摘要报道了一种荧光生物测定法，用于测定碱性磷酸酶 (ALP) 的活性。它正在利用抗坏血酸 (AA) 诱导的由花状羟基氧化钴和铜纳米团簇组成的纳米团簇分解。ALP 催化抗坏血酸 2-磷酸 (AA2P) 的水解和去磷酸化以形成 AA。图形摘要报道了一种荧光生物测定法，用于测定碱性磷酸酶 (ALP) 的活性。它正在利用抗坏血酸 (AA) 诱导的由花状羟基氧化钴和铜纳米团簇组成的纳米团簇分解。ALP 催化抗坏血酸 2-磷酸 (AA2P) 的水解和去磷酸化以形成 AA。