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Method for detecting lysozyme

A detection method, lysozyme technology, applied in the field of biochemical analysis, can solve problems such as narrow linear range, and achieve the effect of high stability and high accuracy

Active Publication Date: 2017-07-18
LINYI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the detection method of lysozyme has electrochemiluminescence method. Although the sensitivity of electrochemiluminescence is high, the linear range of quantification is narrow.

Method used

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  • Method for detecting lysozyme

Examples

Experimental program
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Effect test

preparation example Construction

[0030] The method provided by the invention uses the metal-organic framework material as a modifier for modifying the gold electrode. The metal-organic framework adopted in the invention has a larger specific surface area, and can effectively adsorb the lysozyme aptamer through hydrogen bonds and ππ interactions, thereby increasing the lysozyme Aptamer load. After adding lysozyme, the resulting electron transfer resistance increased significantly. If no metal-organic framework was added, the electrode could not effectively adsorb the lysozyme aptamer, and the electron transfer impedance hardly changed after adding lysozyme. In the present invention, the preparation method of the metal organic framework comprises the following steps: dissolving 2-aminoterephthalic acid in a volatile organic solvent to obtain a 2-aminoterephthalic acid solution; In the phthalic acid solution, a strong alkali solution is added dropwise to adjust the pH value of the mixed solution to 5-6; then th...

Embodiment 1

[0067] Preparation of standard curve:

[0068] Drop-coat 20 μL metal-organic framework solution onto the surface of the gold electrode, incubate at 37°C for 3 h, wash the surface of the gold electrode three times with deionized water, and dry it with nitrogen to obtain a metal-organic framework-modified gold electrode; then drip 20 μL lysozyme aptamer solution Apply to the surface of the metal-organic framework-modified gold electrode, incubate at 37°C for 2 hours, wash the surface of the gold electrode three times with deionized water, and blow dry with nitrogen; use the obtained lysosome aptamer-metal-organic framework-modified gold electrode as the working electrode , using the three-electrode system to perform the first electrochemical impedance spectroscopy detection, and obtain the basic electrical impedance spectrum;

[0069] The steps of polishing are: polish the gold electrode on the suede successively with 0.5μm and 0.05μm alumina solutions, and then use ddH 2 The g...

Embodiment 2

[0077] selective experiment

[0078] Drop-coat 20 μL metal-organic framework solution onto the surface of the polished gold electrode, incubate at 37°C for 3 h, wash the surface of the gold electrode three times with deionized water, and dry it with nitrogen to obtain a metal-organic framework-modified gold electrode; then add 20 μL lysozyme aptamer The solution was drip-coated on the surface of the metal-organic framework-modified gold electrode, incubated at 37 ° C for 2 h, and the surface of the gold electrode was washed with deionized water for 3 times, and dried with nitrogen; the obtained lysosome aptamer-metal-organic framework-modified gold electrode was The working electrode uses a three-electrode system for the first electrochemical impedance spectroscopy detection to obtain a basic electrical impedance spectrum;

[0079] The polishing steps are as follows: the gold electrode is polished on the suede with 0.5 μm and 0.05 μm aluminum oxide solutions in sequence, then ...

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Abstract

The invention provides a method for detecting lysozyme. The method comprises the following steps of dispensing a metal organic skeleton solution to the surface of a gold electrode, carrying out incubating, washing and nitrogen drying; dispensing a lysozyme aptamer solution to the surface of the gold electrode, carrying out incubating, washing and nitrogen drying and carrying out first electrochemical impedance spectroscopy detection; washing the gold electrode after first electrochemical impedance spectroscopy detection, carrying out nitrogen drying, dispensing the solution to a to-be-detected sample, carrying out incubating, washing and nitrogen drying, and carrying out second electrochemical impedance spectroscopy detection; subtracting electron transfer resistance in a first electrical impedance spectroscopy from the electron transfer resistance in a impedance spectroscopy of the to-be-detected sample to obtain an electron transfer resistance difference and comparing with a default standard curve to obtain the concentration of the lysozyme. According to the embodiment of the invention, the adopted detection method is simple in operation, marking is not needed, the flexibility is relatively high, the detection limit can reach 2.0*10<-11>mol / L and the linear range is 1.0*10<-7>mol / L to 1.0*10<-11>mol / L.

Description

technical field [0001] The invention belongs to the technical field of biochemical analysis, in particular to a method for detecting lysozyme. Background technique [0002] Lysozyme, also known as muramidase or N-acetylmuramic glycan hydrolase, is an alkaline enzyme that can hydrolyze mucopolysaccharides in pathogenic bacteria, mainly by destroying N-acetylmuramic acid and N-acetylmuramic acid in the cell wall. -The β-1,4 glycosidic bond between acetylglucosamine decomposes the insoluble mucopolysaccharides of the cell wall into soluble glycopeptides, causing the contents of the cell wall to break and the bacteria to dissolve. Lysozyme can also directly combine with negatively charged viral proteins, and form double salts with DNA, RNA, and apoproteins to inactivate the virus. Therefore, the enzyme has antibacterial, anti-inflammatory, antiviral and other effects. [0003] At present, the detection method of lysozyme includes electrochemiluminescence method. Although the s...

Claims

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Application Information

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IPC IPC(8): G01N27/07G01N27/30
Inventor 虞召朋冯斌时鹏飞
Owner LINYI UNIVERSITY
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