Fluorescence quenching based alkaline phosphatase activity detection

Mei, Yaqi; Hu, Qiong; Zhou, Baojing; Zhang, Yonghui; He, Minhui; Xu, Ting; Li, Feng; Kong, Jinming

doi:10.1016/j.talanta.2017.07.095

Cited by 40 publications

(6 citation statements)

References 62 publications

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“…An increasing number of researchers have designed and synthesized ESIPT-based fluorescent probes for the selective detection of various anions, metal ions, and small organic molecules [16][17][18][19][20][21]. There are also many theoretical studies for the mechanistic elaboration of various fluorescence probes, as well as the design of novel probes [22][23][24][25][26][27][28][29][30][31][32]. Fluorescent probes for ALP detection generally feature phosphate as a recognition moiety, and after cleavage of the phosphate ester group, a phenolic hydroxyl group will be formed.…”

Section: Introductionmentioning

confidence: 99%

Theoretical Investigation on the ESIPT Process and Detection Mechanism for Dual-Proton Type Fluorescent Probe

Cao

Sun

et al. 2022

IJMS

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Recently, a new fluorescent probeAE-Phoswas reported to detect the activity of alkaline phosphatases (ALP) in different living cell lines. Here, we present an in-depth computational analysis of the mechanism and source of the fluorescence of the AE-Phos probe. There is an intermediate product (AE-OH-Phos) in the experiment as well as a different configuration of products that may emit fluorescence. It is essential to investigate the origin of fluorescence and the detection mechanism of the probe, which could help us eliminate the interference of other substances (including an intermediate product and possible isomers) on fluorescence during the experiment. According to the change of geometric parameters and Infrared spectra, we deduce that the dual intramolecular hydrogen bonds of salicylaldehyde azine (SA) were enhanced at the excited state, while AE-OH-Phos was attenuated. Considering the complex ESIPT behavior of the dual proton-type probe, the potential energy surfaces were further discussed. It can be concluded that the single proton transfer structure of SA (SA-SPT) is the most stable form. Both the concerted double proton transfer process and stepwise single proton transfer process of SA were forbidden. The fluorescence for SA was 438 nm, while that of SA-SPT was 521 nm, which agrees with the experimentally measured fluorescence wavelength (536 nm). The conclusion that single proton transfer occurs in SA is once again verified. In addition, the distribution of electron-hole and relative index was analyzed to investigate the intrinsic mechanism for the fluorescence quenching of the probe and the intermediate product. The identification of the origin of fluorescence sheds light on the design and use of dual-proton type fluorescent probes in the future.

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Section: Introductionmentioning

confidence: 99%

Theoretical Investigation on the ESIPT Process and Detection Mechanism for Dual-Proton Type Fluorescent Probe

Cao

Sun

et al. 2022

IJMS

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“…Although these fluorescence probes showed highly selectivity or sensitivity to ALP, they may be affected by factors such as environmental conditions (eg temperature, pH, solvent polarity, etc.) and the concentration of the probe . The ratiometric probe allows measurement of emission at two wavelengths with inherent advantages of high sensitivity and low background, which make them excellent candidates not only in vitro bioassays, but also in the detection of target analytes in vivo. − …”

mentioning

confidence: 99%

“…and the concentration of the probe. 22 The ratiometric probe allows measurement of emission at two wavelengths with inherent advantages of high sensitivity and low background, which make them excellent candidates not only in vitro bioassays, but also in the detection of target analytes in vivo. 23−26 However, only several ratiometric probes for the detection of ALP have been reported.…”

mentioning

confidence: 99%

Ratiometric Fluorescent Strategy for Localizing Alkaline Phosphatase Activity in Mitochondria Based on the ESIPT Process

Zhang

et al. 2019

Anal. Chem.

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Fluorescent probes are powerful tools for detecting and mapping the species of interest in vitro and in vivo. Although the probes always show high selectivity and sensitivity, they are usually affected by some factors, such as detecting conditions and the probe concentrations. Ratiometric fluorescent strategies, possessing advantage of low background noise, would solve the problem effectively and lead to a higher sensing performance. Thus, an ESIPT-based ratiometric probe (HBTP-mito) was developed on the basis of a phosphorylated 2-(2′-hydroxyphenyl)-benzothiazole derivative for the determination of ALP activity. HBTP-mito is water soluble and emits green fluorescence in TBS buffer due to the blockage of ESIPT. Upon the introduction of ALP, the phosphate ester of HBTP-mito was hydrolyzed and the ESIPT process was restored. Accordingly, the fluorescence at 514 nm decreases, while emission at 650 nm shows a “turn-on” response. The ratio of intensity (I 514nm/I 650nm) decreases linearly with ALP activity increasing from 0 to 60 mU/mL, obtained an LOD of 0.072 mU/mL. The favorable performance of the probe enables its application not only in the detection of ALP activity in biological samples, but also in the localization of the ALP levels in living cells and in vivo.

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“…Up to now, a variety of analytical techniques, including electrochemistry, fluorometry, colorimetry, chromatography, and surface-enhanced Raman scattering have been used in the determination of ALP activity [8][9][10][11][12][13][14][15][16][17]. For example, Wu et al established a signal amplified electrochemical method which relied on enzyme-induced metallization (EIM) for ALP analysis [18].…”

Section: Introductionmentioning

confidence: 99%

Sensitive Fluorescence Assay for the Detection of Alkaline Phosphatase Based on a Cu2+-Thiamine System

Zhao

Liu

2021

Sensors

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The authors describe a novel, facile, and sensitive fluorometric strategy based on a Cu2+-thiamine (Cu2+-TH) system for the detection of alkaline phosphatase (ALP) activity and inhibition. The principle of the method is as follows. Under a basic conditions, TH, which does not exhibit a fluorescence signal, is oxidized into fluorescent thiochrome (TC) by Cu2+. Ascorbic acid 2-phosphate (AAP), which is the enzyme substrate, is hydrolyzed to produce ascorbic acid (AA) by ALP. The newly formed AA then reduces Cu2+ to Cu+, which prevents the oxidation of TH by Cu2+; as a result, the fluorescent signal becomes weaker. On the contrary, in the absence of ALP, AAP cannot reduce Cu2+; additions of Cu2+ and TH result in a dramatic increase of the fluorescent signal. The sensing strategy displays brilliant sensitivity with a detection limit of 0.08 U/L, and the detection is linear in the concentration range of 0.1 to 100 U/L. This approach was successfully applied to ALP activity in human serum samples, indicating that it is reliable and may be applied to the clinical diagnosis of ALP-related diseases.

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Fluorescence quenching based alkaline phosphatase activity detection

Cited by 40 publications

References 62 publications

Theoretical Investigation on the ESIPT Process and Detection Mechanism for Dual-Proton Type Fluorescent Probe

Theoretical Investigation on the ESIPT Process and Detection Mechanism for Dual-Proton Type Fluorescent Probe

Ratiometric Fluorescent Strategy for Localizing Alkaline Phosphatase Activity in Mitochondria Based on the ESIPT Process

Sensitive Fluorescence Assay for the Detection of Alkaline Phosphatase Based on a Cu2+-Thiamine System

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