Pharmacy College, Henan University of Chinese Medicine, Zhengzhou, Henan, 450046, People's Republic of China.
Anal Bioanal Chem. 2022 Sep;414(23):6955-6964. doi: 10.1007/s00216-022-04264-4. Epub 2022 Aug 16.
Alkaline phosphatase (ALP), an important hydrolase involved in dephosphorylation, is a common clinical indicator of many diseases. In the present study, we constructed a novel electrochemical sensor using amifostine as the substrate of ALP and activators regenerated by electron transfer atom transfer radical polymerization (ARGET ATRP) as a signal amplification strategy for sensitive determination of ALP activity. In particular, in the presence of ALP, the phosphate group of amifostine was hydrolyzed to form a sulfhydryl group, which could attach to a gold electrode via a sulfur-gold bond. Then, the initiator α-bromophenylacetic acid (BPAA) was linked to the hydrolysis product of amifostine through an amide bond, resulting in the production of electroactive polymer chains on the gold electrode by the monomer ferrocenylmethyl methacrylate (FMMA) via ARGET ATRP. Under optimal parameters, the electrochemical sensor demonstrated a limit of detection (LOD) of 1.71 mU mL with a linear range of 5-100 mU mL. In addition to satisfactory selectivity, the potential application of this approach for ALP activity detection in human serum samples was demonstrated. Due to its efficiency, simplicity of operation, and cost-effectiveness, the proposed electrochemical sensor has great promise as a universal method for ALP assays and inhibitor screening.
碱性磷酸酶（ALP）是一种参与去磷酸化的重要水解酶，是许多疾病的常见临床指标。在本研究中，我们构建了一种新型电化学传感器，使用氨磷汀作为 ALP 的底物，并使用电子转移原子转移自由基聚合（ARGET ATRP）再生的激活剂作为信号放大策略，用于灵敏测定 ALP 活性。特别地，在 ALP 的存在下，氨磷汀的磷酸基团被水解形成巯基，该巯基可通过硫金键附着到金电极上。然后，通过酰胺键将引发剂α-溴代苯乙酸（BPAA）连接到氨磷汀的水解产物上，通过ARGET ATRP，单体二茂铁甲基甲基丙烯酸酯（FMMA）在金电极上产生电活性聚合物链。在最佳参数下，电化学传感器的检测限（LOD）为 1.71 mU mL，线性范围为 5-100 mU mL。除了令人满意的选择性外，还证明了该方法在人血清样品中 ALP 活性检测中的潜在应用。由于其效率、操作简单性和成本效益，所提出的电化学传感器有望成为 ALP 测定和抑制剂筛选的通用方法。
