A novel self-assembled dual-emissive ratiometric fluorescent nanoprobe for alkaline phosphatase sensing

Zhixuan Han; Nan Wang; Yuntai Lv; Qingjie Fu; Guannan Wang; Xingguang Su

doi:10.1016/j.aca.2023.342146

A novel self-assembled dual-emissive ratiometric fluorescent nanoprobe for alkaline phosphatase sensing

Anal Chim Acta. 2024 Jan 25:1287:342146. doi: 10.1016/j.aca.2023.342146. Epub 2023 Dec 17.

Authors

Zhixuan Han¹, Nan Wang¹, Yuntai Lv¹, Qingjie Fu¹, Guannan Wang², Xingguang Su³

Affiliations

¹ Department of Analytical Chemistry, College of Chemistry, Jilin University, Changchun, 130012, China.
² School of Pharmacy, Shenyang Medical University, Shenyang, 110034, China. Electronic address: chemwangguannan@126.com.
³ Department of Analytical Chemistry, College of Chemistry, Jilin University, Changchun, 130012, China. Electronic address: suxg@jlu.edu.cn.

PMID: 38182401
DOI: 10.1016/j.aca.2023.342146

Abstract

Background: Alkaline phosphatase (ALP) is widely found in various organs and tissues of the human body which could assist in the verification of the presence of various diseases through its content in the blood. In the past few years, many analytical methods for ALP activity assays have been explored. However, a simple and economical method with high sensitivity and specificity also remains great challenge. Therefore, the development of sensitive and efficient approach for ALP analysis is of great significance in biomedical studies.

Results: Herein, we constructed a highly sensitive and label-free ratiometric fluorometric biosensing platform for the determination of ALP activity, which utilizing lysozyme(Ly)-functionalized 5-methyl-2-thiouracil(MTU)-modified gold nanoclusters (MTU-Ly@Au NC) and poly-dopamine (PDA) as signal indicators. Dopamine (DA) can self-polymerizes to form PDA under alkaline conditions that can further quenched the fluorescence of MTU-Ly@Au NC at 525 nm due to fluorescence resonance energy transfer (FRET) and absorption competition quenching (ACQ) effects. In this process, the PDA fluorescence intensity at 325 nm was nearly unchanged. After the addition of ALP, ascorbic acid (AA) which can alleviate the self-polymerization process of DA was generated from the substrate ascorbic acid 2-phosphate (AAP), thus changing ratiometric fluorescence intensity of I₅₂₅/I₃₂₅. Hence, by monitoring the fluorescence ratio (I₅₂₅/I₃₂₅), a ratiometric fluorescence biosensing platform for ALP was established with the linear calibration in the range of 0.5-8 U L^-1 and the limit of detection of 0.157 U L^-1.

Significance: This work not only synthesized a novel fluorescence probe with simple preparation and low cost for ALP which has excellent anti-interference properties and selectivity. Furthermore, this biosensing platform was successfully applied for the determination of ALP activity in human serum samples. This work provided a potential tool for biomedical diagnostics in the future.

Keywords: Alkaline phosphatase; Gold nanoclusters; Polydopamine; Ratiometric fluorescence.

MeSH terms

Alkaline Phosphatase* / chemistry
Biological Assay
Calibration
Dopamine
Fluorescent Dyes*
Gold / chemistry
Humans
Metal Nanoparticles / chemistry

Substances

Alkaline Phosphatase
Dopamine
Fluorescent Dyes
Gold