School of Materials Science and Engineering, University of Jinan, Jinan, Shandong 250022, China.
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Scences, Changchun, Jilin 130022, China.
Anal Chem. 2020 Mar 17;92(6):4639-4646. doi: 10.1021/acs.analchem.0c00224. Epub 2020 Mar 5.
Enzyme-triggered in situ chromogenic and/or fluorogenic reactions under accessible conditions are significant for developing enzyme activity and related spectroscopic assays. Here, we describe a facile one-pot synthetic strategy to prepare silicon-containing nanoparticles with yellow-green fluorescence and orange-red color by mixing -[3-(trimethoxysilyl)propyl]ethylenediamine and -aminophenol (AP) in aqueous solution at a mild temperature. Encouraged by the AP-regulated simple synthetic procedure and the generation of AP from alkaline phosphatase (ALP)-catalyzed hydrolysis of 4-aminophenol phosphate (APP), a fluorometric and colorimetric dual-readout ALP activity assay can be rationally envisioned and developed by employing APP as the substrate. In the wake of the good analytical performance of such ALP activity assay and its successful combination with enzyme-linked immunosorbent assay (ELISA), corresponding fluorometric and colorimetric dual-readout ALP-based ELISA has been constructed for highly sensitive and quantitative determination of human prostate-specific antigen (PSA), the key biomarker of prostate cancer in human serum. The convincing performance in evaluating the PSA level in serologic tests unambiguously reveals the great potential of our proposed fluorometric and colorimetric dual-channel immunoassay in early clinical diagnosis by monitoring disease biomarkers.
在可及条件下,酶触发的原位显色和/或荧光反应对于开发酶活性和相关光谱分析至关重要。在这里,我们描述了一种简便的一锅合成策略,即在温和的温度下将-[3-(三甲氧基硅基)丙基]乙二胺和 -氨基酚(AP)混合在水溶液中制备具有黄绿色荧光和橙红色的含硅纳米粒子。受 AP 调节的简单合成程序以及碱性磷酸酶(ALP)催化 4-氨基酚磷酸盐(APP)水解生成 AP 的启发,可以合理地设想并开发基于 APP 作为底物的荧光和比色双读数 ALP 活性测定法。鉴于这种 ALP 活性测定法的良好分析性能及其与酶联免疫吸附测定(ELISA)的成功结合,相应的基于荧光和比色双读数的基于 ALP 的 ELISA 已被构建用于人血清中前列腺癌关键生物标志物人前列腺特异性抗原(PSA)的高灵敏度和定量测定。在评估血清检测中 PSA 水平的令人信服的性能明确显示了我们提出的荧光和比色双通道免疫分析在通过监测疾病生物标志物进行早期临床诊断中的巨大潜力。