Dual-enzyme cascade—An amplified method for the detection of alkaline phosphatase

Mize, Patrick D.; Hoke, Randal A.; Linn, C. P.; Reardon, John E.; Schulte, Thomas

doi:10.1016/0003-2697(89)90120-6

Cited by 28 publications

(9 citation statements)

References 14 publications

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“…Several methods have been utilized to detect ALP activity in the last few decades. − Among them, the fluorescent methods are preferred due to their noninvasiveness, high sensitivity and superb spatiotemporal resolution in visualizing bioreactive species in the biological systems. − Until now, a few fluorescent probes have been reported for monitoring ALP activity through various mechanisms, such as assembly of nanoparticles, host–guest interaction, and change of solubility. − However, the application of these probes is hampered by their intrinsic drawbacks, such as low sensitivity, laborious synthetic procedures, complicated sensing mechanism as well as short emission and excitation wavelengths. Recently, fluorescent probes based on aggregation-induced emission (AIE) mechanism have been reported for detecting ALP in serum and living cells. − Although these probes showed elevated sensitivity and selectivity, the requirement for high probe concentration (50 μM) and short wavelength emission can limit their application in biological systems.…”

Section: Introductionmentioning

confidence: 99%

Reaction-Based Off–On Near-infrared Fluorescent Probe for Imaging Alkaline Phosphatase Activity in Living Cells and Mice

Tan

Zhang

Man³

et al. 2017

ACS Appl. Mater. Interfaces

134

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Alkaline phosphatases are a group of enzymes that play important roles in regulating diverse cellular functions and disease pathogenesis. Hence, developing fluorescent probes for in vivo detection of alkaline phosphatase activity is highly desirable for studying the dynamic phosphorylation in living organisms. Here, we developed the very first reaction-based near-infrared (NIR) probe (DHXP) for sensitive detection of alkaline phosphatase activity both in vitro and in vivo. Our studies demonstrated that the probe displayed an up to 66-fold fluorescence increment upon incubation with alkaline phosphatases, and the detection limit of our probe was determined to be 0.07 U/L, which is lower than that of most of alkaline phosphatase probes reported in literature. Furthermore, we demonstrated that the probe can be applied to detecting alkaline phosphatase activity in cells and mice. In addition, our probe possesses excellent biocompatibility and rapid cell-internalization ability. In light of these prominent properties, we envision that DHXP will add useful tools for investigating alkaline phosphatase activity in biomedical research.

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Section: Introductionmentioning

confidence: 99%

Reaction-Based Off–On Near-infrared Fluorescent Probe for Imaging Alkaline Phosphatase Activity in Living Cells and Mice

Tan

Zhang

Man³

et al. 2017

ACS Appl. Mater. Interfaces

134

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“…Indirect procedures have also been developed in which the phenol acts as a substrate or an inhibitor in a coupled enzymatic indicator reaction Mize et al, 1989). The assay described here illustrates a new assay principle in which the liberated phenol acts as an enhancer or anti-enhancer of a chemiluminescent enzymatic indicator reaction (Figure 1).…”

Section: Discussionmentioning

confidence: 99%

“…Substituted phenol derivatives are widely used to assay hydrolase enzymes and the liberated phenol is detected directly by virtue of its color, fluorescence (McComb et al, 1979), or chemiluminescence .' Indirect procedures have also been developed in which the phenol acts as a substrate or an inhibitor in a coupled enzymatic indicator reaction Mize et al, 1989). The assay described here illustrates a new assay principle in which the liberated phenol acts as an enhancer or anti-enhancer of a chemiluminescent enzymatic indicator reaction (Figure 1).…”

Section: Discussionmentioning

confidence: 99%

Chemiluminescent assay of enzymes using proenhancers and pro‐anti‐enhancers

Kricka

Schmerfeld-Pruss

Edwards³

1991

J. Biolumin. Chemilumin.

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Enhanced chemiluminescent assays for hydrolase enzymes have been developed using proenhancer and pro-anti-enhancer substrates. Alkaline phosphatase is measured using disodium para-iodophenyl phosphate (proenhancer) which is converted to para-iodophenol and this in turn enhances the light emission from the horseradish peroxidase catalysed chemiluminescent oxidation of luminol by peroxide. An alternative strategy uses para-nitrophenyl phosphate which is converted by alkaline phosphatase to para-nitrophenol which inhibits the enhanced chemiluminescent reaction. The detection limit for the enzyme using the proenhancer and pro-anti-enhancer assays was 100 attomoles and 1 picomole, respectively. The proenhancer strategy was effective in assays for beta-D-galactosidase, beta-D-glucosidase and aryl sulfatase. A limited comparison of the proenhancer and a conventional colorimetric assay for an alkaline phosphatase label in an enzyme immunoassay for alpha-fetoprotein showed good agreement.

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“…Typically, the signals produced by these assays are directly proportional to the analyte concentration . However, linear signal‐gain systems such as these exhibit significantly worse sensitivity and dynamic range than exponential amplification systems, which are much less prevalent in the literature . Additionally, these systems often require enzyme reporters and sensitive spectroscopic instrumentation to achieve maximum performance, limiting their applicability to a narrow range of laboratory settings.…”

Section: Figurementioning

confidence: 99%

Peroxidyme‐Amplified Radical Chain Reaction (PARCR): Visible Detection of a Catalytic Reporter

Goertz

White

2017

Angewandte Chemie

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Peroxidyme Amplified Radical Chain Reaction (PARCR), anovel enzyme-free system that achieves exponential amplification of av isible signal, is presented. Typical enzyme-free amplification systems that produce avisible readout suffer from long reaction times,low sensitivity,and narrow dynamic range.P ARCR employs photocatalyzed nonlinear signal generation, enabling unprecedented one-pot, naked-eye detection of acatalytic reporter from 1 mm down to 100 pm.In this reaction, hemin-binding peroxidase-mimicking DNAzymes ("peroxidymes") mediate the NADH-driven oxidation of acolorless,nonfluorescent phenoxazine dye (Amplex Red) to abrightly colored, strongly fluorescent product (resorufin); illumination with green light initiates multiple radical-forming positive-feedbackl oops,r apidly producing visible levels of resorufin. Collectively,t hese results demonstrate the potential of PARCR as an easy-to-use readout for ar ange of detection schemes,i ncluding aptamer labels,h ybridization assays,a nd nucleic acid amplification.

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Dual-enzyme cascade—An amplified method for the detection of alkaline phosphatase

Cited by 28 publications

References 14 publications

Reaction-Based Off–On Near-infrared Fluorescent Probe for Imaging Alkaline Phosphatase Activity in Living Cells and Mice

Reaction-Based Off–On Near-infrared Fluorescent Probe for Imaging Alkaline Phosphatase Activity in Living Cells and Mice

Chemiluminescent assay of enzymes using proenhancers and pro‐anti‐enhancers

Peroxidyme‐Amplified Radical Chain Reaction (PARCR): Visible Detection of a Catalytic Reporter

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