Alkaline phosphatase is one of the most important tool enzymes and diseases indicator, monitoring ALP activity with convenient, precise, efficient and sensitive methods plays a fundamental role in modern life and healthcare industries. In this study, we described a novel method for ALP analysis based on Pb²⁺ dependent DNAzyme. By modifying DNAzyme sequence with terminal phosphate group and introducing exonuclease I (exo I), we managed to analyze ALP by utilizing its causal function of DNAzyme probe from exo I mediated degradation and function of triggering the subsequent cleavage of the hairpin reporting probe. Other than one amplificative strategy by DNAzyme mediated cleavage and cycle, this system also involved an exo I mediated degradation to further reduce the background noise. Combining stepwise fluorimetry and electrophoresis, we verified the detective mechanism of this proposed method. Further, after selectivity demonstration, this method achieved a considerable LOD of 0.0017 U L⁻¹ and linear range of 0.0025 U L⁻¹ to 250 U L⁻¹. For potential of practical application, this method also exhibited excellent performances in inhibitor screening and intracellular ALP assay, both with a linear fitting equation. Based on these results, this method should be highly committed for improving ALP analysis in modern life industry.

碱性磷酸酶是最重要的工具酶和疾病指示剂之一，以方便、精确、高效、灵敏的方法监测ALP活性在现代生活和医疗保健行业发挥着基础性作用。在这项研究中，我们描述了一种基于 Pb ^{2+的 ALP 分析新方法}依赖性脱氧核糖核酸酶。通过用末端磷酸基团修饰 DNAzyme 序列并引入外切核酸酶 I (exo I)，我们设法利用 DNAzyme 探针从 exo I 介导的降解的因果功能和触发随后的发夹报告探针切割的功能来分析 ALP。除了通过 DNAzyme 介导的切割和循环的一种扩增策略外，该系统还涉及 exo I 介导的降解，以进一步降低背景噪音。结合逐步荧光法和电泳法，我们验证了该方法的检测机制。此外，在选择性论证之后，该方法实现了相当大的 0.0017 UL ^{-1的 LOD 和 0.0025 UL -1}至 250 UL ^-1的线性范围. 对于实际应用的潜力，该方法在抑制剂筛选和细胞内 ALP 测定中也表现出优异的性能，两者均具有线性拟合方程。基于这些结果，该方法应高度致力于改进现代生活行业的ALP分析。