ANALYTICA CHIMICA ACTA
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2016
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IF 4.712
Shenghao Xu,Xiuying Feng,Teng Gao,Ruizhi Wang,Yaning Mao,Jiehua Lin,Xijuan Yu,Xiliang Luo
A novel ultrasensitive dual-functional biosensor for highly sensitive detection of inorganic pyrophosphate (PPi) and pyrophosphatase (PPase) activity was developed based on the fluorescent variation of globulin protected gold nanoclusters (Glo@Au NCs) with the assistance of Cu 2+ . Glo@Au NCs and PPi were used as the fluorescent indicator and substrate for PPase activity evaluation, respectively. In the presence of Cu 2+ , the fluorescence of the Glo@Au NCs will be quenched owing to the formation of Cu 2+ -Glo@Au NCs complex, while PPi can restore the fluorescence of the Cu 2+ -Glo@Au NCs complex because of its higher binding affinity with Cu 2+ . As PPase can catalyze the hydrolysis of PPi, it will lead to the release of Cu 2+ and re-quench the fluorescence of the Glo@Au NCs. Based on this mechanism, quantitative evaluation of the PPi and PPase activity can be achieved ranging from 0.05 μM to 218.125 μM for PPi and from 0.1 to 8 mU for PPase, with detection limits of 0.02 μM and 0.04 mU, respectively, which is much lower than that of other PPi and PPase assay methods. More importantly, this ultrasensitive dual-functional biosensor can also be successfully applied to evaluate the PPase activity in human serum, showing great promise for practical diagnostic applications.
研究领域:
生物化学
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分析化学
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医学诊断
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产品关联
DOI
PubMed
高影响因子
Nanomaterials
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2019
·Vol. 9
·期 1
·111
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IF 4.034
Yishen Tian,Lijie Hao,Chao Wang,Xiaoyan Yang,Shufeng Liu
Development of simple, convenient, and sensitive assay methods for pyrophosphatase (PPase) activity is of importance, for disease diagnosis and drug discovery. Herein, a simple, rapid, label-free, and sensitive fluorescence sensor for PPase activity assay is developed, using Cu2+doping-induced quantum dot (QD) photoluminescence as a signal reporter. The Cu2+doping of ZnSe QD can induce a dopant-dependent emission response, which will be inhibited after the premixing of Cu2+with pyrophosphate (PPi), to form a Cu2+-PPi complex. Then, the hydrolysis of PPi into phosphate (Pi), specifically catalyzed by PPase, liberates the free Cu2+to regain the QD doping for the fluorescence response, which is highly dependent on the PPase activity. The PPase can be sensitively and selectively assayed, with a detection limit of 0.1 mU/mL. The developed sensing strategy can be also employed for the PPase inhibitor screening. Thus, the current QD doping-based sensing strategy offers an efficient and promising avenue for Cu2+, PPi, or PPase-related target analysis, and might hold great potential for the further applications in the clinical disease diagnosis.Keywords:quantum dot doping;pyrophosphatase activity;fluorescence detection;pyrophosphate;copper ion
研究领域:
生物化学
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医学诊断
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纳米技术
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DOI
PubMed
高影响因子
Journal of Analytical Methods in Chemistry
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2016
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IF 1.369
Lingzhi Zhao,Liu Zhao,Yanqing Miao,Chunye Liu,Chenxiao Zhang
The detection of pyrophosphatase (PPase) activity is of great significance in diagnosing diseases and understanding the function of PPase-related biological events. This study constructed a turn off-on-off fluorescent system for PPase activity assay based on PPase-regulated competitive coordination of Cu2+ between a water-soluble fluorescent probe 6,7-dihydroxycoumarin (DHC) and pyrophosphate (PPi). The probe DHC can coordinate with Cu2+ and consequently display on-off type fluorescence response. Furthermore, the in situ formed nonfluorescent Cu2+-DHC complex can act as an effective off-on type fluorescent probe for sensing PPi due to the higher coordination reactivity between Cu2+ and PPi than that between Cu2+ and DHC. The subsequent addition of PPase to the mixture containing Cu2+, DHC, and PPi leads to the fluorescence requenching of the system again (an off state) because PPase catalyzes the hydrolysis of PPi into orthophosphate in the reaction system. Under the optimum conditions, the decrease of the fluorescence intensity of DHC-Cu2+-PPi system was linear with the increase of the PPase activity in the range from 0.1 to 0.3 U. The detection limit was down to 0.028 U PPase (). Moreover, the as-established system was also applied to evaluate PPase inhibitor. This study offers a simple yet effective method for the detection of PPase activity.
研究领域:
生物化学
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医学检测
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
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2016
·
IF 5.886
Jian Sun,Bin Wang,Xue Zhao,Zong-Jun Li,Xiurong Yang
We demonstrate a rationally designed fluorescent and colorimetric dual-readout strategy for the highly sensitive, quantitative determination of inorganic pyrophosphatase (PPase) activity, a key hydrolytic enzyme involved in a variety of metabolic processes. Inspired by the selective oxidative and chromogenic reaction of o-phenylenediamine (OPD) with Cu2+, the special inhibitory effects of pyrophosphate (PPi) on the oxidative ability of Cu2+, and the specific hydrolysis of PPi into orthophosphate by PPase, a convenient small molecule OPD-based analytical system was developed for Cu2+/PPi recognition and PPase activity assay. We have confirmed that Cu2+ acts as the oxidant in the reaction and the main chromogenic product of OPD is 2,3-diaminophenazine (usually called OPDox) in the assay by combining the ESI-MS, 1H NMR, and XPS spectra analysis. Direct electrochemical insights into the Cu2+-triggered and PPi-inhibited mechanism were performed by cyclic voltammetry characterizations of the Cu2+ in the absence and presence of PPi for the first time. Furthermore, the proposed analytical system with clear response mechanism exhibits a promising outlook for the PPase activity assay in real biological samples and inhibitor screening.
研究领域:
生物化学
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分析化学
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
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2018
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IF 5.123
Xuechao Xu,Xiaobo Zou,Shuwen Wu,Linjie Wang,Xiangheng Niu,Xin Li,Jianming Pan,Hongli Zhao,Minbo Lan
As pyrophosphate ion (PPi) and pyrophosphatase (PPase) play crucial roles in the pathological process of arthritis, determination of PPi and PPase in biological fluids turns to be of great importance for clinical diagnosis and therapy of arthritic diseases. In this work, we proposed a new fluorescent assay for PPi and PPase activity detection based on the competitive coordination chemistry of Fe 3+ between PPi and an in situ formed fluorescent polydopamine (PDA). FeCo layered double hydroxide (FeCo-LDH) was explored as a peroxidase mimic to facilitate the in situ formation of fluorescent PDA from dopamine mediated by low-concentration H 2 O 2 within 30 min; The formed fluorescent PDA could be significantly quenched by Fe 3+ through forming a PDA-Fe 3+ complex structure; When PPi existed, it coordinated Fe 3+ competitively against PDA and inhibited the fluorescence quenching of PDA by Fe 3+ ; When PPi was hydrolyzed under the catalysis of PPase, the Fe 3+ ion could quench the fluorescence of the formed PDA again. With these principles, our fluorescent assay was able to detect PPi and PPase activity specifically, providing detection limits down to 54 μM and 0.13 U/L, respectively. Furthermore, accurate determination of PPi and PPase activity in spiked human serum was also demonstrated using the developed assay.
研究领域:
生物化学
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医学诊断
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
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2014
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IF 5.825
Jian Sun,Fan Yang,Dan Zhao,Xiurong Yang
On the basis of the competition assay approach and fluorescent 11-mercaptoundecanoic acid-capped AuNCs (AuNCs@11-MUA) with unique optical properties, a convenient, reliable and highly sensitive real-time assay of pyrophosphatase (PPase) activity is established and developed for the first time. Pyrophosphate (PPi) could recover the Cu2+-quenched AuNCs@11-MUA fluorescence selectively owing to the higher binding affinity between PPi and Cu2+ than that between 11-MUA and Cu2+. Whereas PPase could catalyze the hydrolysis of PPi, thus released Cu2+, leading to fluorescence requenching of the AuNCs@11-MUA. In the assay, a good linearity between the fluorescence response and PPase activity within a range from 1 to 20 mU is found, with a detection limit of less than 1 mU, which is better than other PPase assays using PPi as the substrate. Additionally, we demonstrate that our AuNCs@11-MUA-based fluorescent assay can be applied to assay the PPase activity in real biological samples such as the cell lysate. This strategy paves a new avenue for exploring the sensing applications of fluorescence AuNCs and improving the development of competition assay approach.
研究领域:
生物化学
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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
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2022
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IF 4.831
Ningning Zhang,Lixin Zhao,Mengting He,Peng Luo,Liang Tan
A highly sensitive and selective sensor for the quantitative assay of inorganic pyrophosphatase (PPase) activity was developed based on a fluorescence “turn–off” strategy. Carbon quantum dots@Cu(II)-based metal–organic framework nanotubes (CQDs@Cu-MOF) with length less than 300 nm and width less than 20 nm were synthesized. CQDs in the nanotubes exhibited weak fluorescence owing to static quenching. The coordination reaction between pyrophosphate ion (PPi) and Cu(II) decomposed CQDs@Cu-MOF and led to the release of CQDs, of which the fluorescence recovered. In the presence of PPase, the hydrolysis of PPi generated phosphate ion (Pi). CQDs@Cu-MOF remained their structural stability and the fluorescence turned off. The fluorescence intensity difference of the mixture of CQDs@Cu-MOF and PPi in the absence and presence of PPase (-Δ F ) was proportional to the PPase concentration from 0.1 to 5 mU mL −1 and that from 5 to 50 mU mL −1 , and a limit of detection at 0.03 mU mL −1 was obtained. PPase activity in human serum was analyzed using the proposed fluorescence sensor and the recovery values were found to vary from 95.0% to 104 %.
研究领域:
生物化学
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分析化学
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产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
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2019
·Vol. 186
·期 3
·1-8
·
IF 5.479
Hu Shuisheng,Zhu Lin,Lam Cheong Wing,Guo Longhua,Lin Zhenyu,Qiu Bin,Wong Kwok Yin,Chen Guonan,Liu Zhenhua
A copper(II)-based two-dimensional metal-organic framework with nanosheet structure (CuBDC NS) that possesses peroxidase (POx) mimicking activity was prepared. In the presence of hydrogen peroxide, the system catalyses the oxidation of terephthalic acid to a blue-fluorescent product (excitation = 315 nm; emission = 425 nm). Pyrophosphate has a very strong affinity for Cu 2+ ion and blocks the POx-mimicking activity of the CuBDC NS. If, however, inorganic pyrophosphatase is present, the POx mimicking activity is gradually restored because pyrophosphate is hydrolyzed. The findings were used to design a method for the determination of the activity of inorganic pyrophosphatase by fluorometry. Fluorescence increases linearly in the 1–50 mU·mL −1 inorganic pyrophosphatase activity range. The limit of detection is 0.6 mU·mL −1 (S/ N = 3).
A copper(II)-based two-dimensional metal-organic framework (CuBDC NS) is described that possesses POx-mimicking activity. Inorganic pyrophosphate (PPi) was hydrolyzed to phosphate in the presence of inorganic pyrophosphatase (PPase). Hence, it cannot coordinate with Cu 2+ in CuBDC NS, its structure was well-conserved to catalyses the oxidation of terephthalic acid (H 2 BDC) to produce a blue fluorescent product (oxBDC) in the presence of hydrogen peroxide (H 2 O 2 ).
研究领域:
生物化学
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分析化学
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DOI
PubMed
高影响因子
Frontiers in Bioengineering and Biotechnology
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2021
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IF 5.89
Lei Zhongli,Zhou Jie,Liang Miao,Xiao Yan,Liu Zhihong
The development of sensitive and accurate detection of inorganic pyrophosphate (PPi) and pyrophosphatase activity (PPase) is important as they play vital roles in biological systems. However, it is still not satisfactory for most of the analytical methods for PPi and PPase because of their Cu2+-dependence and poor accuracy. Although the metal ion triggered aggregation-induced emission (AIE) of metal nanoclusters (NCs) offers a new approach to design a Cu2+-free strategy for the accurate determination of PPi and PPase recently, current methods are all focused on utilizing pure metal NCs. Alloy NCs incorporating the advantages of diverse metal usually can achieve improved behaviors in the application, such as enhanced sensitivity and stability. In this work, glutathione stabilized alloy Au/Ag NCs were synthesized via a simple method and used for the fluorescence detection of PPi and PPase based on a Zn2+-regulated AIE strategy. The controlled release of Zn2+ by PPi and PPase could regulate the AIE of Au/Ag NCs and be employed to response PPi concentration and PPase activity. This method processes simple procedure, high sensitivity and stability, and low toxicity. In addition, we also studied the AIE behaviors of this Au/Ag NCs and offer some fundamental understanding of the AIE properties of water-soluble alloy NCs. This study not only provides a straightforward and new approach for PPi and PPase determination but a basis for further study on the AIE properties of alloy NCs and their application.
研究领域:
分析化学
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生物化学
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DOI
PubMed
高影响因子
MICROCHIMICA ACTA
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2020
·Vol. 187
·期 12
·1-8
·
IF 6.232
Pang Jiawei,Lu Yuexiang,Gao Xinyu,He Liuying,Sun Jingwei,Yang Fengyi,Liu Yueying
A fluorescence method for the determination of inorganic pyrophosphatase (PPase) activity has been established based on copper nanoclusters (CuNCs). The polythymine of 40 mer (T40) acts as a template for the reduction reaction from Cu 2+ to Cu 0 by ascorbic acid (AA). This reaction leads to the formation of fluorescent CuNCs with excitation/emission peaks at 340/640 nm. However, the higher binding affinity between inorganic pyrophosphate (PPi) and Cu 2+ hinders the effective formation of CuNCs. This shows low fluorescence intensity. PPase catalyzes the hydrolysis of PPi into Pi during which free Cu 2+ ions are produced. This facilitates the formation of fluorescent CuNCs. Thus, the fluorescence intensity was restored. The fluorescence enhancement of the system has a linear relationship with PPase activity in the range 0.3 to 20 mU·mL −1 , and the detection limit is0.2 mU·mL −1 . The relative intensity (I/I 0 ) at 640 nm for the analytical solution versus system is also employed to screen the inhibitor for PPase with high efficiency.
Schematic representation of a fluorescent assay for the determination of inorganic pyrophosphatase activity and screening its inhibitor based on single-strand polythymine-scaffolded copper nanoclusters.
研究领域:
生物化学
·
分析化学
·
分子生物学
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2019
·
IF 5.256
Xuan Zhao,Wentao Li,Tong Wu,Pingping Liu,Wei Wang,Guiyun Xu,Shenghao Xu,Xiliang Luo
Herein, a zinc ion (Zn 2+ )-triggered aggregation induced emission enhancement (AIEE) fluorescence “on-off-on” nanoswitch was fabricated for inorganic pyrophosphate (PPi) and inorganic pyrophosphatase (PPase) activity detection. Dual ligand functionalized Au NCs were utilized as the substrate of the AIEE nanoswitch. The introduction of Zn 2+ can cause Au NCs aggregated along with the enhanced fluorescence. After the addition of PPi, aggregated Au NCs disaggregated along with decreased fluorescence due to the competitive combination between PPi and Zn 2+ (on-off). When PPase was introduced, PPi was hydrolyzed and release Zn 2+ , resulting in aggregated Au NCs along with enhanced fluorescence again (off-on). On the basis of this, highly selective and sensitive detection PPi (liner range from 0.1 to 300 μM) and PPase activity (liner range from 0.1 to 10 mU) can be achieved. The detection limits are 0.04 μM for PPi and 0.03 mU for PPase, respectively. Furthermore, the as-prepared Zn 2+ -triggered AIEE nanoswitch was successfully used for quantitative analysis of PPase activity in human serum with satisfactory spiked recoveries, and applied for the inhibitors screening.
研究领域:
分析化学
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生物医学工程
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产品关联
DOI
PubMed
高影响因子
Chemosensors
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2021
·Vol. 9
·期 3
·48
·
IF 3.398
Wei Wang,Hongren Zhao,Bing Zhao,Huimin Liu,Qinglei Liu,Yan Gao
In this paper, a novelNL-Fe3+ensemble was designed as a fluorescent chemosensor for highly selective detection of pyrophosphate (PPi) in DMSO/H2O (2:8/v:v, pH = 7.2) solution and living cells.NLshowed a strong affinity for Fe3+and was accompanied by obvious fluorescence quenching. Upon the addition of PPi to the generatedNL-Fe3+ensemble, the fluorescence and absorption spectra were recovered completely. Spectroscopic investigation showed that the interference provoked by common anions such as adenosine-triphosphate (ATP), adenosine diphosphate (ADP), and phosphates (Pi) can be ignored. The detection limit ofNL-Fe3+to PPi was calculated to be 1.45 × 10−8M. Intracellular imaging showed thatNL-Fe3+has good membrane permeability and could be used for the detection of PPi in living cells. A B3LYP/6-31G(d,p) basis set was used to optimizeNLandNL-Fe3+complex.Keywords:coumarin;Fe(III);pyrophosphate;cells imaging
研究领域:
生物化学
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分析化学
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细胞生物学
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DOI
高影响因子
危险品化学品经营许可证（带存储）