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A fluorescent probe for measuring acetylcholinesterase and its preparation method and application

An acetylcholinesterase and fluorescent probe technology, which is applied in the field of enzyme activity detection, can solve the problems of complex instruments, limit practical application and the like, and achieve the effect of simple and easy preparation method.

Active Publication Date: 2022-02-18
QINGDAO UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Nowadays, the detection of acetylcholinesterase mainly relies on Ellman method, electrochemical method, surface-enhanced resonance Raman scattering method, colorimetric method and fluorescence method. Most of the above methods require complex instruments, which limits their practical application.

Method used

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  • A fluorescent probe for measuring acetylcholinesterase and its preparation method and application
  • A fluorescent probe for measuring acetylcholinesterase and its preparation method and application
  • A fluorescent probe for measuring acetylcholinesterase and its preparation method and application

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preparation example Construction

[0043] A method for preparing a fluorescent probe for measuring acetylcholinesterase, the specific steps of the method are as follows:

[0044] Add probe master solution and CuCl in PBS buffer solution at room temperature 2 Solution, stirring and coordination reaction to obtain the target product HBTP-Cu 2+ Complex fluorescent probes.

[0045] Preferably, the pH value of the PBS buffer solution is controlled between 7.5 and 8.5.

[0046] Preferably, the probe mother liquor and the CuCl 2 The molar ratio of the solution is 2:1, and the concentration of the probe mother solution is 2.0~3.0mmol / L, the CuCl 2 The concentration of the solution is 0.05-0.20 mol / L.

[0047] A HBTP-Cu 2+ Application of complex fluorescent probe to selectively recognize acetylcholinesterase in solvent system.

[0048] In order to further realize the technical effect of the present invention, in the solvent system, the volume percentage of water is 95-100%.

[0049] A HBTP-Cu 2+ The application of...

Embodiment 1

[0052] HBTP-Cu 2+ The preparation method of the complex: Add the probe mother solution (8 μL, 2.0 mmol·L -1 ) and CuCl 2 Solution (0.1μL, 0.05mol L -1 , 0.5equ.), that is, mixed according to the molar ratio of 2:1, coordination occurs, and HBTP-Cu is obtained 2+ Aqueous solution of the complex.

Embodiment 2

[0054] HBTP-Cu 2+ The preparation method of the complex: add the probe mother solution (8 μL, 2.5 mmol L to 2 mL PBS buffer solution (pH=7.8) -1 ) and CuCl 2 Solution (0.1μL, 0.1mol L -1 , 0.5equ.), that is, mixed according to the molar ratio of 2:1, coordination occurs, and HBTP-Cu is obtained 2+ Aqueous solution of the complex.

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Abstract

The invention discloses a fluorescent probe for measuring acetylcholinesterase and its preparation method and application. The fluorescent probe is HBTP-Cu 2+ Compound, and the HBTP-Cu disclosed by the present invention 2+ The complex is composed of HBTP and thiocholine (acetylcholine is specifically catalyzed by acetylcholinesterase) and Cu 2+ The competitive binding of HBTP is used as the basic mechanism of action, and HBTP luminescence is used as the fluorescent signal reporter group. When acetylcholinesterase is added to the system, thiocholine can interact with HBTP‑Cu 2+ Cu in the composite structure 2+ The complex structure is disintegrated by the coordination combination, and the existing form of HBTP changes from ordered aggregation to monomer dispersion in the bulk solution, and changes in the color and fluorescence of the solution occur. The recognition effect on acetylcholinesterase was studied by nuclear magnetic resonance, ultraviolet-visible absorption spectroscopy, and fluorescence spectroscopy, and the results showed that HBTP-Cu 2+ The complex can efficiently and selectively recognize acetylcholinesterase in a pure water solvent system, and has high sensitivity to acetylcholinesterase.

Description

technical field [0001] The invention belongs to the technical field of enzyme activity detection, and relates to a method strategy for detecting endogenous acetylcholinesterase in cells. More specifically, it relates to a fluorescent probe, its preparation method and its application in detecting acetylcholinesterase. Background technique [0002] Acetylcholinesterase (AChE), a cholinesterase that controls carboxypeptidase and aminopeptidase functions, selectively catalyzes the hydrolysis of the substrate acetylcholine (ATh), a neurotransmitter for nerve signaling. AChE helps promote neuronal development and nerve regeneration, and is mainly distributed in tissues in the human body consisting of the brain, liver, and spinal cord. In addition, there is growing evidence that abnormally elevated alkaline phosphatase levels in the blood are associated with various diseases such as cancer, heart disease, bone disease, and liver disease. Therefore, it is of great significance to ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D417/10C09K11/06G01N21/64
CPCC07D417/10C09K11/06G01N21/6428C09K2211/1037
Inventor 丁彩凤傅彩霞张倩张鹏
Owner QINGDAO UNIV OF SCI & TECH
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