Real-time fluorescence assays of alkaline phosphatase and ATP sulfurylase activities based on a novel PPi fluorescent probe

Wang, Xiaobo; Zhang, Zhiyang; Ma, Xiaoyan; Wen, Jinghan; Geng, Zhirong; Wang, Zhilin

doi:10.1016/j.talanta.2015.01.028

Cited by 28 publications

(19 citation statements)

References 34 publications

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“…Among the various methods , fluorescent methods present many advantages, including high sensitivity, low cost, easy detection, and suitability as a diagnostic tool for biological concern. In recent years, several fluorescent substrates for ALP have been developed , such as salicylic acid phosphate , 5‐fluorosalicylic acid phosphate , β‐naphthol phosphate , trifluoromethyl‐β‐umbelliferone phosphate , 2‐carboxy‐1‐naphthyl phosphate , and 8‐quinolyl phosphate . Although the fluorescent probe is highly sensitive, it may be influenced by some factors such as photobleaching, changes of probe concentration, and environmental conditions (pH, polarity, temperature, and so forth).…”

Section: Introductionmentioning

confidence: 99%

“…canceling artifacts due to probe concentration variations) and increase the selectivity, sensitivity and dynamic ranges of the method . However, up to now, only a few ratiometric fluorescent probes for ALP have been reported and they suffered the drawbacks in complicated process in probe synthesis. Therefore, there is a strong desire to develop ratiometric fluorescent probes for ALP with new mechanism.…”

Section: Introductionmentioning

confidence: 99%

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A ratiometric fluorescent probe for alkaline phosphatase via regulation of excited‐state intramolecular proton transfer

Fan

Luo

2015

Luminescence

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A ratiometric fluorescent probe 2-(benzimidazol-2-yl)phenyl phosphoric acid (1) for alkaline phosphatase (ALP) is designed and synthesized. The method employs the modulation of the excited-state intramolecular proton transfer (ESIPT) process of 2-(2'-hydroxyphenyl)benzimidazole (HPBI) through the hydroxyl group protection/deprotection reaction. Upon phosphorylated with POCl3 , HPBI shows only an emission peak at 363 nm due to the blockage of ESIPT. However, once selective enzymatic hydrolysis with alkaline phosphatase (ALP) in Tris-HCl buffer occurs, the probe 1 is returned to HPBI and the ESIPT process is switched on, which results in a decrease in the emission band at 363 nm and an increase in a new fluorescence peak around 430 nm. The fluorescence intensity ratio at 430 and 360 nm (I430/I360) increases linearly with the activity of ALP up to 0.050 U/mL and the detection limit is 0.0013 U/mL. The proposed probe shows excellent specificity toward ALP.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A ratiometric fluorescent probe for alkaline phosphatase via regulation of excited‐state intramolecular proton transfer

Fan

Luo

2015

Luminescence

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“…We also investigated the inhibition effect of the ALP inhibitor. 42 Sodium orthovanadate (Na3VO4), a frequently used inhibitor of ALP, was used to study the inhibition effect, 43 by changing the concentration of the inhibitor from 0 to 2.5 mM with 200 mU mL -1 ALP and keeping all other situations the same. The absorption spectrum of each group is presented in Fig.…”

Section: Inhibitor Screeningmentioning

confidence: 99%

Metal-to-Ligand Charge-Transfer-based Visual Detection of Alkaline Phosphatase Activity

Mei

et al. 2018

ANAL. SCI.

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The ability to directly detect alkaline phosphatase (ALP) activity in undiluted serum samples is of great importance for clinical diagnosis. In this work, we report the use of the distinctive metal-to-ligand charge-transfer (MLCT) absorption properties of the Cu(BCA) (BCA = bicinchoninic acid) reporter for the visual detection of ALP activity. In the presence of ALP, the substrate ascorbic acid 2-phosphate (AAP) can be enzymatically hydrolyzed to release ascorbic acid (AA), which in turn reduces Cu to Cu. Subsequently, the complexation of Cu with the BCA ligand generates the chromogenic Cu(BCA) reporter, accompanied by a color change of colorless-to-purple of the solution with a sharp absorption band at 562 nm. The underlying MLCT-based mechanism has been demonstrated on the basis of density functional theory (DFT) calculations. Needless of any sequential multistep operations and elaborately designed colorimetric probe, the proposed MLCT-based method allows for a fast and sensitive visual detection of ALP activity within a broad linear range of 20 - 200 mU mL (R = 0.999), with a detection limit of 1.25 mU mL. The results also indicate that it is highly selective and has great potential for the screening of ALP inhibitors in drug discovery. More importantly, it shows a good analytical performance for the direct detection of the endogenous ALP levels of undiluted human serum samples. Owing to the prominent simplicity and practicability, it is reasonable to conclude that the proposed MLCT-based method has a high application prospect in clinical diagnosis.

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“…8,9 These observations have been attributed to the free motion of the rotor structures in solution consuming excited state energy by nonradiative decay to quench the emissionand the blocking of the intramolecular rotational motion and intramolecular vibration in the aggregated state activating the radiative decay pathway to yield strong uorescence. 10,11 Molecules that undergo excited-state intramolecular proton transfer (ESIPT) display a larger Stokes shi, which can effectively prevent self-absorption. Hence, a great deal of effort has been put into studying the optical properties of molecules displaying ESIPT with AIE.…”

Section: Introductionmentioning

confidence: 99%

Nano-aggregates of furan-2-carbohydrazide derivatives displaying enhanced emission with a bathochromic shift

Ding

Wang

et al. 2019

RSC Adv.

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Real-time fluorescence assays of alkaline phosphatase and ATP sulfurylase activities based on a novel PPi fluorescent probe

Cited by 28 publications

References 34 publications

A ratiometric fluorescent probe for alkaline phosphatase via regulation of excited‐state intramolecular proton transfer

A ratiometric fluorescent probe for alkaline phosphatase via regulation of excited‐state intramolecular proton transfer

Metal-to-Ligand Charge-Transfer-based Visual Detection of Alkaline Phosphatase Activity

Nano-aggregates of furan-2-carbohydrazide derivatives displaying enhanced emission with a bathochromic shift

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