Analytical Methods
·
2014
·Vol. 6
·期 15
·6105-6109
·
IF 1.938
Fangyuan Wang,Yongxin Li,Wenying Li,Qingfeng Zhang,Jian Chen,Huipeng Zhou,Cong Yu
In the present study, a facile fluorescence turn-on approach for the detection of alkaline phosphatase (ALP) activity has been developed. L-ascorbic acid-2-phosphate (AAP), a substrate of ALP, could be hydrolyzed by ALP to give L-ascorbic acid. L-ascorbic acid then reduced resazurin to resorufin, which resulted in a turn-on fluorescence signal. The fluorescence intensity increase could be directly related to the amount of ALP added to the assay solution. The assay is very sensitive, as 0.12 mU mL−1 ALP could be clearly detected. Sodium orthovanadate (Na3VO4), a well-known ALP inhibitor, was tested, and a clear inhibition effect was observed. The results suggest that our method could be used for ALP activity sensing related various biochemical applications and for the screening of ALP inhibitors.
研究领域:
生物化学
·
分析化学
·
分子生物学
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DOI
高影响因子
MICROCHIMICA ACTA
·
2017
·Vol. 185
·期 1
·1-6
·
IF 4.58
Mao Mi,Tian Tian,He Yu,Ge Yili,Zhou Jiangang,Song Gongwu
Boron and nitrogen codoped carbon dots functionalized with cyclodextrin (β-CD-N/B-C-dots) were obtained from β-cyclodextrin. The material displays strong fluorescence (with excitation/emission peak wavelengths of 400/500 nm) and was characterized by UV-vis, transmission electron microscopy and FTIR. If the substrate p-nitrophenylphosphate is enzymatically cleaved by alkaline phosphatase (ALP), a yellow product is formed whose absorption overlaps the excitation spectrum of the β-CD-N/B-C-dots. Hence, fluorescence is reduced due to an inner filter effect. In additon, the β-CD cavity offers a pocket for substrate recognition. The findings were used to design a method for the determination of the activity of ALP. It has a working range that extends from 0.003 to 5.5 U·L −1 , with a 0.3 mU·L −1 detection limit. The method is fast, simple, inexpensive, and highly sensitive and selective.
Schematic of an inner filter effect based probe for alkaline phosphatase based on the use boron and nitrogen co-doped carbon dots (N/B-C-dots) modified with β-cyclodextrin (β-CD). PNPP: p-Nitrophenylphosphate; PNP: p-Nitrophenol anion.
研究领域:
生物化学
·
分析化学
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DOI
PubMed
高影响因子
Analytical Methods
·
2019
·Vol. 11
·期 42
·5472-5477
·
IF 2.378
Linjie Wang,Kun Ye,Jianming Pan,Hongwei Song,Xin Li,Xiangheng Niu
As a typical biomarker for disease diagnosis in clinical practice, alkaline phosphatase (ALP) plays a critical role in regulating the dephosphorylation process of biomolecules. On this account, exploration of an effective and simple means of ALP activity measurement becomes very significant. In this work, we report a catalytic reaction-based colorimetric method for the monitoring of ALP activity via employing nanostructured MnO2 as a favorable oxidase mimic. The prepared MnO2 microspheres exhibit good oxidase-like capacity to induce the catalytic oxidation of colorless 3,3′,5,5′-tetramethylbenzidine (TMB) to deep-blue TMBox; when ALP hydrolyzes ascorbic acid phosphate, the produced ascorbic acid (AA) with a certain reducing power can destroy the MnO2 enzyme mimic by reducing it to Mn2+, dramatically suppressing the TMB chromogenic reaction. Based on the strategy, a convenient and high-performance colorimetric method was established for ALP activity sensing, giving a wide linear range from 0.5 to 120 U L−1. With the help of N-ethylmaleimide as an efficient masking agent for biothiols, the colorimetric approach could provide excellent specificity for ALP activity detection. Reliable and precise measurement of the analyte in practical samples was also verified by our assay, indicating its promise as a feasible tool for clinical diagnosis.
研究领域:
生物化学
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医学诊断
·
纳米材料科学
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DOI
高影响因子
Talanta Open
·
2024
·
IF 4.1
Chen-Xi Zhao,Xiao-Xia Li,Yang Shu
Abnormal alkaline phosphatase levels have been associated with several diseases, including tumor growth, diabetes, liver dysfunction and so on. Generally, the occurrence and prognosis of these diseases can be reflected by the serum alkaline phosphatase level. In this work, Eu/DBM@SiO 2 @SiO 2 is prepared to protect the internal Eu complex from the external environment by wrapping Eu/DBM inside the silica shell. Then, Eu/DBM@SiO 2 @SiO 2 /MnO 2 , a sensor consists of Eu/DBM@SiO 2 @SiO 2 and MnO 2 , is prepared and applied for alkaline phosphatase detection. MnO 2 is used as the response part of alkaline phosphatase, which can be degraded to Mn 2+ by ascorbic acid. Ascorbic acid is the hydrolysate of alkaline phosphatase. In the process of this reaction, UV–vis absorption of Eu/DBM@SiO 2 @SiO 2 /MnO 2 is reduced and fluorescence is turned on. Alkaline phosphatase concentration is linearly correlated with the logarithmic value of fluorescence enhancement in the range of 10.0–100.0 U/L with linear equation of Lg(F/F 0 )=0.008C-0.030 (R 2 =0.99). For fluorescence method, the LOQ is 10.0 U/L and the LOD is 4.4 U/L. For colorimetric method, the concentration of alkaline phosphatase is linearly correlated with the decrease of absorption intensity in the range of 20.0–90.0 U/L with the linear equation of A/A 0 =-0.012C+1.148 (R 2 =0.99). The LOQ is 20.0 U/L and the LOD is 7.2 U/L. The detection system has good selectivity and can detect alkaline phosphatase in human serum samples, the accuracy of the detection system is verified by the experiment of spike-in experiment. The RSD of alkaline phosphatase concentration in serum samples is in the range of 1.4–8.4 %. This method has potential application in the detection of ALP activity in biological samples.
研究领域:
生物化学
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医学诊断
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DOI
高影响因子
Journal of Materials Chemistry B
·
2025
·
IF 5.7
Yingxian Shi,Ruirui Dong,Huan Huang,Gaoying Wang,Jianyi Gao,Kai Wang,Ting Zhang
Ovarian cancer (OC) remains a prevalent gynecological malignancy, posing significant threats to women's health globally. Emerging evidence underscores the critical involvement of alkaline phosphatase (ALP) in OC pathogenesis, prompting the development of targeted diagnostic tools. Herein, we report the successful application of a novel fluorescent probe, HD-ALP, for fluorescent imaging of ALP in ovarian cancer cells and preclinical models. After being triggered by ALP, HD-ALP responded in the fluorescent signal reporting manner. More importantly, the fluorescence signal showed a reliable linear correlation with ALP levels in the range of 0-10000 U/L. The HD-ALP probe demonstrated high sensitivity, selectivity, and stability under the necessary biological conditions. The intracellular imaging results indicated that HD-ALP enabled the visualization of ALP levels within ovarian cancer cells. The in vivo experiments demonstrated that HD-ALP could accurately differentiate between healthy and cancerous models (subcutaneous transplantation tumor model of ovarian cancer, orthotopic ovarian cancer model, primary ovarian cancer model). Notably, In cellular and in vivo imaging, HD-ALP could effectively reflect the decrease in fluorescence signal after the addition of ALP inhibitors. By integrating ALP detection, which utilizes the HD-ALP fluorescence mode, with traditional methods like immunohistochemistry staining and serum biomarker detection, potential strategies for more accurate diagnosis of OC may be developed for preclinical trials in the future. According to the research, this is also one of the few developments of fluorescent detection tools for measuring ALP levels in ovarian cancer models.
研究领域:
肿瘤学
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医学影像学
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生物化学
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DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2024
·
IF 8.3
Jieqiong Chen,Qing Zhao,Jiajing Tang,Xiaoyu Lei,Jinzheng Zhang,Yuping Li,Jidong Li,Yubao Li,Yi Zuo
Inspired by the crucial role of matrix vesicles (MVs), a series of biomimetic vesicles (BVs) fabricated by calcium glycerophosphate (CaGP) modified polyurethane were designed to mediate the mineralization through in situ enzyme activation for bone therapy. In this study, alkaline phosphatase (ALP) was harbored in the porous BVs by adsorption (Ad-BVs) or entrapment (En-BVs). High encapsulation of ALP on En-BVs was effectively self-activating by calcium ions of CaGP-modified PU that specifically hydrolyzed the organophosphorus (CaGP) to inorganic phosphate, thus promoting the formation of the highly oriented bone-like apatite in vitro. Enzyme-catalyzed kinetics confirms the regulation of apatite crystallization by the synergistic action of self-activated ALP and the confined microcompartments of BVs. This leads to a supersaturated microenvironment, with the En-BVs group exhibiting inorganic phosphate (Pi) levels 4.19 times higher and Ca2+ levels 3.67 times higher than those of simulated body fluid (SBF). Of note, the En-BVs group exhibited excellent osteo-inducing differentiation of BMSCs in vitro and the highest maturity with reduced bone loss in rat femoral defect in vivo. This innovative strategy of biomimetic vesicles is expected to provide valuable insights into the enzyme-activated field of bone therapy.
研究领域:
生物材料学
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骨再生医学
·
组织工程
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2025
·
IF 7.7
Xiangyu Cai,Yuying Jia,Xuepeng Wang,Hongyan Shan,Yanfu Huan
In this work, a novel chemiluminescence (CL) reagent dioleamide (DOA) was synthesized, which contains a long chain organic compound with an amide as the core functional group. It exhibited stable and strong CL emission when sodium hypochlorite was used as the oxidant. A mechanistic study indicates that CL is mainly due to the process in which the reactive oxygen species (ROS) produced by the system during the chemical reaction oxidize the DOA to the excited state and then return to the ground state to release energy in the form of light radiation. Alkaline phosphatase (ALP) catalyzes the hydrolysis reaction of p-nitrophenyl phosphate (PNPP) to generate p-nitrophenol (PNP). The maximum absorption wavelength of PNP significantly overlaps with the CL emission wavelength of the DOA, enabling energy transfer from DOA to PNP and thus the CL intensity is weakened. Consequently, a simple, efficient, and sensitive ALP sensor based on chemiluminescence resonance energy transfer (CRET) system was developed with a detection limit of 0.020 U/L. The NaClO-DOA-PNP sensing system, which demonstrated a highly sensitive reaction to ALP in the range of 0.1–30 U/L, was successfully applied to the detection of ALP in fetal bovine serum.
研究领域:
生物化学
·
分析化学
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产品关联
DOI
高影响因子
LANGMUIR
·
2018
·
IF 3.789
Lele Zhou,Qiongzheng Hu,Qi Kang,Ming Fang,Li Yu
The detection of l-phenylalanine (l-Phe) has become one of the most pressing issues concerning diagnosis and treatment of phenylketonuria in neonates; however, a simple and robust methodology is yet to be developed. Here, the application of novel liquid crystals (LCs)-sensing platform for sensitive, selective, and label-free detection of l-Phe was reported at the first time. We devised a strategy to fabricate the sodium monododecyl phosphate (SMP)-decorated LC sensing platform with the appearance of dark. Then, a dark to bright (D–B) optical images alteration of LCs was observed after transferring alkaline phosphatase (ALP) to the interface, owing to cleavage of SMP induced by ALP. LCs remained dark images after the SMP-decorated interface in contact with the pre-incubated ALP and l-Phe. Such optical appearance resulted from the inhibition of ALP by l-Phe, which was further verified by the isothermal titration calorimetry (ITC). The strategy was applied to sensing l-Phe, which have been proven to allow for sensitively and selectively differentiation of l-Phe from interfering compounds with similar aromatic groups, as well as seven other essential amino acids. More importantly, the detection limit of l-Phe reached 1 pg/mL in urine samples, further demonstrating its value in the practical applications. Results obtained in this study clearly demonstrated the superiority of LCs toward the l-Phe detection, which can pave a way for the development of high performance and robust probes for l-Phe detection in clinical applications.
研究领域:
生物化学
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医学诊断
·
材料科学
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DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2022
·
IF 5.304
Jin-Hong Sui,Yun-Yun Wei,Jin Li,Zhang-Run Xu
Mucin 1 (MUC1) is an important tumor biomarker, and the determination of MUC1 is of great significance in clinical diagnostics, prognosis assessment and therapy monitoring. Developing point-of-care testing method of MUC1 is necessary in resource-limited settings. Herein, a visual multicolor aptasensor for MUC1 detection was designed based on an alkaline phosphatase (ALP)-triggered cascade reaction. In the presence of MUC1, ALP-decorated complementary DNA and MUC1 aptamer on magnetic beads were dissociated due to the recognition between MUC1 and aptamer. With the assistance of a magnet, ALP-decorated complementary DNA was separated and collected to catalyze the hydrolysis of l -ascorbic acid-2-phosphate generating l -ascorbic acid, which etched the manganese dioxide nanosheets to mediate the oxidation reaction of 3,3′,5,5′-tetramethylbenzidine sulfate, resulting in different products with distinct visible multicolor. The naked eye detection of MUC1 was realized and the detection limit was estimated to be 0.18 U mL −1 by using the colorimetric method. This proposed aptasensor possesses the merit of vivid multicolor change, and provides a new way for MUC1 detection by naked eyes.
研究领域:
生物医学工程
·
临床诊断学
·
分子生物学
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DOI
高影响因子
Nanoscale
·
2019
·Vol. 11
·期 29
·13714-13719
·
IF 6.97
Yuhan Wang,Jie Zhan,Yumiao Chen,Sifan Ai,Liantao Li,Ling Wang,Yang Shi,Junnian Zheng,Zhimou Yang
The selective formation of nanomedicines around cancer cells is very important for cancer therapy because it increases the inhibitory capacity and decreases the systemic toxicity. However, successful examples are rare. Taking advantage of the overexpression of both the enzyme alkaline phosphatase (ALP) and the cell membrane receptor (CCK2R), we demonstrated in this study the selective formation of supramolecular nanofibers and hydrogels in the pericellular space of two cancer cell lines (HeLa and HepG2 cells). Both cell lines showed high expression levels of extracellular ALP and membrane-bound CCK2R. ALP efficiently converted Comp. 1 to a self-assembling molecule (Comp. 2). Comp. 2 interacted with CCK2R, thereby facilitating the self-assembly and formation of hydrogels around the cancer cells. The selective pericellular hydrogelations efficiently inhibited cancer cells. Pericellular hydrogelation around cancer cells is a promising strategy to control the formation of nanomedicines spatiotemporally in cellular microenvironments for cancer therapy and diagnostics.
研究领域:
生物医学
·
肿瘤学
·
药剂学
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DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2019
·Vol. 186
·期 3
·1-8
·
IF 5.479
Ni Pengjuan,Xie Junfeng,Chen Chuanxia,Jiang Yuanyuan,Lu Yizhong,Hu Xun
The authors describe a fluorometric method for determination of the activity of alkaline phosphatase (ALP) and its inhibitors. Nitrogen and boron co-doped carbon dots (C-dots) with excitation/emission peaks at 490/540 nm act as the fluorescent probe. The C-dots were prepared by hydrothermal carbonization starting from 3-aminophenylboronic acid as the sole precursor. On the basis of the boronic acid-triggered specific reaction with cis -diols, the boronic acid modified C-dots can bind to ascorbic acid that is generated by ALP-catalyzed hydrolysis of ascorbic acid 2-phosphate. This results in particle aggregation and quenching of fluorescence. If the ALP inhibitor Na 3 VO 4 is introduced into the system, the activity of ALP is reduced and the fluorescence of C-dots recovers. This fluorometric method allows for the determination of ALP activity in the range from 0.2 to 6.0 mU mL −1 with a detection limit of 0.16 mU mL −1 . The IC 50 value for the inhibitor Na 3 VO 4 is 3.6 μM. The method is convenient and cost-effective. It does not require complicated operations and in our perception widens the scope of applications of C-dots in bioanalytical sciences.
Schematic presentation of the nitrogen and boron co-doped carbon dot-based fluorometric method for determination of alkaline phosphatase (ALP) activity.
研究领域:
生物化学
·
分析化学
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产品关联
DOI
PubMed
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2021
·
IF 4.098
Min Huang,Fenghua Geng,Yongxiang Wang,Congying Shao,Gen Liu,Maotian Xu
The photometric method is widely used in real clinical tests due to its simple operation, low cost and convenient. Many of the reported colorimetric ALP assays so far are non- ratiometric because the detection was based on changes in absorbance at a single wavelength. The development of novel colorimetric and ratiometric assay is of importance for quantitatively measuring target with high accuracy. The challenge in the design of ratiometric photometric assay is that the chromophore must have a significant spectral shift before and after binding to the target. Here, we report a colorimetric and ratiometric photometric sequential assay of AA and ALP based on the complexation between ARS and Cu 2+ and redox reaction between AA and Cu 2+ . The absorption band of ARS centered at 425 nm (yellow color), which could be shifted to 510 nm (red color) upon Cu 2+ binding. However, as far as we know, this classic color reaction has not been used to develop a ratiometric photometric method to sequentially detect AA and ALP, although photometric methods based on the regulation of other color reagents with oxidizing metal ions have been reported. The proposed sensing system shows a limit of detection for ALP at 0.24 U L −1 and could be applied for detecting ALP in newborn calf serum. The established sensing system makes a useful contribution to the detection of ALP in complex clinical samples.
研究领域:
临床检验
·
生物化学
·
医学诊断
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）