Ministry of Education Key Laboratory for Analytical Science of Food Safety and Biology, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, College of Chemistry, Fuzhou University, Fuzhou, Fujian, 350116, China.
Pharmaceutical and Medical Technology College of Putian University, Putian, Fujian, China.
Analyst. 2022 Jun 13;147(12):2749-2756. doi: 10.1039/d2an00517d.
Alkaline phosphatase (ALP) plays a vital role in clinical diagnoses and biomedical research. It is important to develop some convenient but sensitive methods for ALP activity detection. In this study, a multicolor biosensor for ALP activity has been developed based on the peroxidase activity of copper nanoclusters (CuNCs) and etching of gold nanorods (AuNRs). The presence of CuNCs can catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) to produce blue TMB. In acid solution, TMB can cause the etching of AuNRs accompanied by a significant color change of the system. The presence of sodium pyrophosphate (PPi) can inhibit the peroxidase activity of CuNCs, which can be recovered after the addition of ALP. Different ALP added results in the recovery of the catalytic activity of CuNCs to different degrees and generates different amounts of TMB. This consequently affected the morphology of the AuNRs in the system and results in the output of a vivid color change, which can be recognized with the naked eyes easily without any complicated instruments. The biosensor has a linear relationship with ALP activity in the range of 10.0-80.0 U L, and the detection limit is 4.6 U L. The proposed method has been applied to detect ALP activity in human serum samples with satisfactory results.
碱性磷酸酶(ALP)在临床诊断和生物医学研究中起着至关重要的作用。开发一些方便但灵敏的 ALP 活性检测方法非常重要。在本研究中,基于铜纳米簇(CuNCs)的过氧化物酶活性和金纳米棒(AuNRs)的刻蚀作用,开发了一种用于 ALP 活性的多色生物传感器。CuNCs 的存在可以催化 3,3',5,5'-四甲基联苯胺(TMB)的氧化,生成蓝色 TMB。在酸性溶液中,TMB 可以引起 AuNRs 的刻蚀,同时伴随着体系颜色的明显变化。焦磷酸钠(PPi)的存在可以抑制 CuNCs 的过氧化物酶活性,加入 ALP 后可以恢复其活性。不同量的 ALP 加入会导致 CuNCs 的催化活性恢复程度不同,生成的 TMB 量也不同。这会导致体系中 AuNRs 的形态发生变化,从而产生鲜明的颜色变化,无需任何复杂仪器,用肉眼即可轻松识别。该生物传感器在 10.0-80.0 U L 的范围内与 ALP 活性呈线性关系,检测限为 4.6 U L。该方法已成功应用于人血清样品中 ALP 活性的检测,结果令人满意。