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Metal-organic framework-based β-glucuronidase probes and applications

A metal-organic framework and glucuronide technology, which is applied in the field of analytical chemistry and clinical testing, can solve the problems of complex operation, time-consuming and cost, and achieve the effect of simple operation, high sensitivity and fast response

Active Publication Date: 2022-06-07
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the current enzyme fluorescence detection methods require the synthesis and preparation of suitable substrate molecules or organic fluorescent probes, which are not only complicated, time-consuming and costly

Method used

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  • Metal-organic framework-based β-glucuronidase probes and applications
  • Metal-organic framework-based β-glucuronidase probes and applications
  • Metal-organic framework-based β-glucuronidase probes and applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] In this example, the probe substrate is 4-methylumbelliferone-β-glucuronide, and a fluorescence spectrometer is used to detect GCU activity in human serum

[0045] The specific method is as follows: take 50 μL serum, 100 μL 20 μM 4-methylumbelliferone-β-glucuronide, 100 μL 75 mM calcium chloride aqueous solution, plus a certain amount of buffer (pH=5.3, 20 mM Bis-Tris -Glycine buffer) to bring the total solution volume to 1 mL, place the solution in a thermostatic shaker (800 rpm), incubate at 37 degrees Celsius for 100 minutes, then transfer to a quartz cuvette and measure with a fluorescence spectrometer. Fluorescence emission spectra were recorded at an excitation wavelength of 322 nm. The fluorescence intensity at 450 nm was recorded. The response standard curve of GCU activity was established with the fluorescence intensity as the ordinate.

[0046] It can be seen from the results that the detection limit of β-glucuronidase activity by the method described in the...

Embodiment 2

[0048] In this example, the probe substrate is 4-methylumbelliferone-β-glucuronide, and Eu is added. 3+ @MOF-253, GCU activity in human serum detected by fluorescence spectrometer

[0049] The specific method is as follows: take 50 μL serum, mix 100 μL 20 μM 4-methylumbelliferone-β-glucuronide, 55 μL 1 mg / mL Eu 3+ @MOF-253 aqueous solution, 100 μL of 75 mM calcium chloride aqueous solution, plus a certain amount of buffer (pH=5.3, 20 mM Bis-Tris-glycine buffer) to bring the total solution volume to 1 mL, place the solution on a constant temperature shaker (800rpm), incubate at 37°C for 100 minutes, then transfer to a quartz cuvette and measure with a fluorescence spectrometer. Fluorescence emission spectra were recorded at an excitation wavelength of 322 nm. Finally, the ratio of fluorescence intensity at 450 nm and 617 nm was calculated.

[0050] The inspection process reference is attached image 3 , the response standard curve of GCU activity established directly with r...

Embodiment 3

[0056] In this example, the probe substrate is 4-methylumbelliferone-β-glucuronide, and Eu is added. 3+ @MOF-253, Detecting GCU activity in human serum with a luminometer

[0057] The specific method is as follows: take 50 μL serum, mix 100 μL 20 μM 4-methylumbelliferone-β-glucuronide, 55 μL 1 mg / mL Eu 3+ @MOF-253 aqueous solution, 100 μL of 75 mM calcium chloride aqueous solution, plus a certain amount of buffer (pH=5.3, 20 mM Bis-Tris-glycine buffer) to make the total solution volume 1 mL, put the solution on a constant temperature shaker (800rpm), incubate at 37 degrees Celsius for 100 minutes, then transfer to a quartz cuvette, and perform RGB colorimetric analysis with an illuminometer under the excitation of 302nm ultraviolet light in a dark box, thereby realizing quantitative detection of GCU activity.

[0058] The inspection process reference is attached image 3 , the response standard curve of GCU activity established with ratio chromaticity as the ordinate is as f...

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Abstract

The invention provides a metal-organic framework-based β-glucuronidase probe, the probe substrate of the metal-organic framework-based β-glucuronidase probe can be β-glucuronidase Enzyme-specific catalysis, the probe substrate is 4-methylumbelliferone-β-glucuronide. It can realize ratiometric fluorescence and / or ratiometric colorimetric detection of β-glucuronidase activity. Its detection limit is low, which can reach 0.03U / L; it has good selectivity and is less affected by interference; it can realize high-sensitivity naked-eye visual detection (red to blue); at the same time, it has simple operation, fast response, good precision, High accuracy and other outstanding advantages.

Description

technical field [0001] The patent of the present invention relates to the technical field of analytical chemistry and clinical testing, in particular to a β-glucuronidase activity probe, colorimetric analysis device and application based on a metal organic framework. Background technique [0002] β-Glucuronidase (GCU) is a matrix-degrading enzyme involved in the process of tumor invasion and metastasis, and it is widely used as a biomarker for clinical diagnosis of diseases such as cancer (malignant tumors of the pancreas, breast, liver, colon, and cervix). Therefore, it is necessary to establish a high-sensitivity, high-accuracy, and highly portable assay method for β-glucuronidase activity. [0003] A variety of methods have been reported for the detection of β-glucuronidase activity, including colorimetric methods, electrochemical methods, and fluorescence methods. Among them, fluorescence detection has high sensitivity and selectivity, and is widely used in enzyme activ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64G01N21/25
CPCG01N21/6486G01N21/25
Inventor 肖玉秀李朔
Owner WUHAN UNIV
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