Abstract |
Alkaline phosphatase (ALP), an enzyme that catalyzes the hydrolysis of phosphate groups, is closely associated with many diseases, including bone disease, prostate cancer, and diabetes. Thus, new assays for ALP detection in live cells are needed to better understand its role in related biological processes. In this study, we constructed a novel near-infrared ratiometric fluorescent probe for detecting ALP activity with high sensitivity. The probe uses a new self-immolative mechanism that can achieve a rapid response (within 10 min) to ALP, detected as a spectral shift (from 580 to 650 nm). This method effectively avoids issues related to instrument variability, and the near-infrared fluorescence emission (650 nm) makes it more suitable for biological detection. Moreover, the high sensitivity (14-fold enhancement of the fluorescence ratio F650/F580) and low detection limit (0.89 U L-1) for ALP allows the probe to be adapted to complex biological environments. The assay was successfully performed using serum samples with a linear range of ALP of up to 150 U L-1. We used the developed probe to detect and image endogenous ALP in cells with satisfactory results, and we successfully used the probes to detect changes in endogenous ALP levels in zebrafish caused by drug-induced organ damage.
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Authors | Xueyan Zhang, Xiangzhu Chen, Kaizheng Liu, Yuanyuan Zhang, Gui Gao, Xiaoqian Huang, Shicong Hou |
Journal | Analytica chimica acta
(Anal Chim Acta)
Vol. 1094
Pg. 113-121
(Jan 15 2020)
ISSN: 1873-4324 [Electronic] Netherlands |
PMID | 31761037
(Publication Type: Journal Article)
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Copyright | Copyright © 2019 Elsevier B.V. All rights reserved. |
Chemical References |
- Carbamates
- Fluorescent Dyes
- Organophosphates
- Acetaminophen
- Carbon Tetrachloride
- Alkaline Phosphatase
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Topics |
- Acetaminophen
(pharmacology)
- Alkaline Phosphatase
(analysis)
- Animals
- Carbamates
(chemical synthesis, chemistry, toxicity)
- Carbon Tetrachloride
(toxicity)
- Cattle
- Density Functional Theory
- Fluorescent Dyes
(chemical synthesis, chemistry, toxicity)
- HeLa Cells
- Humans
- Limit of Detection
- Microscopy, Confocal
(methods)
- Microscopy, Fluorescence
(methods)
- Models, Chemical
- Organophosphates
(chemical synthesis, chemistry, toxicity)
- Zebrafish
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