Capsulation of EBTAC into ZIF-8 for the development of a signal-on fluorescent biosensor to detect alkaline phosphatase

Abstract

Diseases such as liver cancer, extrahepatic biliary obstruction and osteocarcinoma are closely associated with the abnormal level of alkaline phosphatase (ALP). Hence, it is essential to develop a convenient assay to detect ALP activity. Herein, a novel signal-on fluorescent biosensor on account of the fluorescence signal of the aggregation-induced emission (AIE) fluorochrome 2,2′,2′′,2′′′-((ethene-1,1,2,2-tetrayltetrakis(benzene-4,1-diyl))tetrakis(oxy))tetraacetic acid (EBTAC) encapsulated zeolitic imidazolate framework-8 (ZIF-8@EBTAC) was designed to monitor ALP. Due to the aggregation-induced emission of EBTAC, the synthetic ZIF-8@EBTAC shows robust fluorescence. Once pyrophosphate (ppi) was added, its complexation with Zn²⁺ in ZIF-8 triggered the collapse of the ZIF-8 framework, releasing encapsulated EBTAC molecules and restoring to free state, leading to the dramatical decrease in fluorescence. ALP could catalyze the hydrolysis of ppi to phosphate (pi), which is difficult to bind to Zn²⁺ and has little effect on the fluorescence of ZIF-8@EBTAC. Therefore, with the assistance of the substrate ppi, the ultimate fluorescence of ZIF-8@EBTAC was positively related with ALP activity. The constructed biosensor was able to monitor the ALP activity well from 0.01 to 100 U L⁻¹, and a detection limit of 0.01 U L⁻¹ was achieved. Based on the ability of EBTAC serving as a fluorescent probe with aggregation-induced luminescence properties, this proposed design can be applied to diverse targets and provide new ideas for the establishment of fluorescent biosensors.