State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, 130022, P. R. China.
College of Animal Sciences, Jilin University, Changchun, 130062, P. R. China.
Analyst. 2019 Mar 21;144(6):2034-2041. doi: 10.1039/c8an02231c. Epub 2019 Jan 31.
A novel sensing strategy for acetylcholinesterase (AChE) and alpha-fetoprotein (AFP) is developed, based on the perylene probe monomer to excimer fluorescence transformation induced by the in situ generation of a metal coordination polymer. In the presence of AChE, acetylthiocholine chloride was hydrolyzed to thiocholine. Ag and the produced thiocholine formed a positively charged metal coordination polymer, which induced the aggregation of a negatively charged perylene probe and the formation of probe excimer emission. The intensity ratio of excimer to monomer emission was proportional to the AChE concentration. A sensing method for AChE was thus established with a detection limit of 0.02 mU mL. The excimer emission with a large Stokes shift could avoid the interference of background fluorescence from complex biological samples, and thus achieved selective and sensitive detection of AChE. In addition, a fluorescence immunoassay strategy for AFP was then developed. Gold nanoparticles (AuNPs) co-immobilized with acetylcholinesterase and the AFP antibody as the capture and amplification probe were first prepared. In the presence of AFP, the sandwich structure was formed by immunological recognition. The hydrolysis of acetylthiocholine was catalyzed by AChE on the AuNPs, and the metal coordination polymer was then formed which resulted in the aggregation of the perylene probe and the formation of the excimer emission. The proposed sensing method offers a new strategy for the detection of other biomarkers.
基于芘探针单体的分子内激基缔合物荧光转变,开发了一种用于乙酰胆碱酯酶(AChE)和甲胎蛋白(AFP)的新型传感策略。在 AChE 的存在下,乙酰硫代胆碱被水解为硫代胆碱。Ag 和生成的硫代胆碱形成带正电荷的金属配位聚合物,诱导带负电荷的芘探针聚集并形成探针激基缔合物发射。激基缔合物发射与单体发射的强度比与 AChE 浓度成正比。因此,建立了一种检测 AChE 的传感方法,其检测限为 0.02 mU mL。具有大斯托克斯位移的激基缔合物发射可以避免复杂生物样品背景荧光的干扰,从而实现了对 AChE 的选择性和灵敏检测。此外,还开发了一种用于 AFP 的荧光免疫分析策略。首先制备了与乙酰胆碱酯酶和 AFP 抗体共固定的金纳米粒子(AuNPs)作为捕获和放大探针。在 AFP 的存在下,通过免疫识别形成夹心结构。AuNPs 上的 AChE 催化乙酰硫代胆碱的水解,然后形成金属配位聚合物,导致芘探针聚集并形成激基缔合物发射。所提出的传感方法为检测其他生物标志物提供了一种新策略。