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Acetylcholin esterase biosensor based on metal organic framework as well as preparation method and application of acetylcholin esterase biosensor

A technology of acetylcholinesterase and metal-organic framework, applied in the field of acetylcholinesterase biosensors and its preparation, can solve the problems of complex sample pretreatment, time-consuming and cumbersome process, expensive instruments, etc., and achieve good thermal and chemical stability, good Application prospect, effect of good enzyme activity

Inactive Publication Date: 2019-10-18
YANSHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these technical methods are sensitive and reliable, complex pretreatment of samples is required before detection, which is time-consuming and cumbersome, and the cost is high, and most of the instruments required by these methods are expensive, bulky, and require skilled operators, which limits the environment. On-line Rapid Detection of Food Contamination

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  • Acetylcholin esterase biosensor based on metal organic framework as well as preparation method and application of acetylcholin esterase biosensor
  • Acetylcholin esterase biosensor based on metal organic framework as well as preparation method and application of acetylcholin esterase biosensor
  • Acetylcholin esterase biosensor based on metal organic framework as well as preparation method and application of acetylcholin esterase biosensor

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Embodiment 1

[0033] This embodiment provides a metal-organic framework-based acetylcholinesterase biosensor, the preparation method of which comprises;

[0034] (1) Preparation of acetylcholinesterase@ZIF-8 material:

[0035] First, mix 250 μL of acetylcholinesterase with an activity of 0.25 U / μL and 200 μL of 0.60M zinc acetate and shake for 15 minutes, then add 1800 μL of 3.55M 2-methylimidazole solution into the above mixture, and react in 20-25 Reaction at ℃ for 0.5h to obtain acetylcholinesterase@ZIF-8 biocomposite material;

[0036] The preparation method of ZIF-8 as a control is the same as the preparation method of acetylcholinesterase@ZIF-8 biocomposite material, except that no enzyme is added.

[0037] (2) Preparation of acetylcholinesterase @ZIF-8 material @Au nano-ion:

[0038] The product obtained in step (1) was washed and centrifuged three times, then reconstituted with 500 μL of water, 80 μL of 10 mM chloroauric acid solution was added to the above material, and an equal ...

Embodiment 2

[0046] This embodiment provides a metal-organic framework-based acetylcholinesterase biosensor, the preparation method of which comprises;

[0047] (1) Preparation of acetylcholinesterase@ZIF-8 material:

[0048] First, mix 150 μL acetylcholinesterase with an activity of 0.25 U / μL and 150 μL 0.47M zinc acetate and shake for 15 minutes, then add 1000 μL 2.48M 2-methylimidazole solution to the above mixture, pass through the normal temperature aqueous solution at 25-35 °C After 4.5 hours of reaction, the acetylcholinesterase@ZIF-8 biocomposite material was obtained.

[0049] The prepared acetylcholinesterase@ZIF-8 biocomposite was characterized by laser confocal scanning microscopy, and the results were as follows image 3 As shown, it can be clearly seen from the figure that acetylcholinesterase is wrapped in the ZIF-8 framework.

[0050] (2) Preparation of acetylcholinesterase @ZIF-8 material @Au nano-ion:

[0051] The product obtained in step (1) was washed and centrifuged...

Embodiment 3

[0056] This embodiment provides a metal-organic framework-based acetylcholinesterase biosensor, the preparation method of which comprises;

[0057] (1) Preparation of acetylcholinesterase@ZIF-8 material:

[0058] First, mix 90 μL acetylcholinesterase with an activity of 0.25U / μL and 50 μL 0.31M zinc acetate and shake for 30 minutes, then add 500 μL 1.25M 2-methylimidazole solution to the above mixture, and react at 15-25°C through a normal temperature aqueous solution. Under reaction for 3.5h, the acetylcholinesterase@ZIF-8 biocomposite material was obtained;

[0059] The preparation method of ZIF-8 is the same as the preparation method of acetylcholinesterase@ZIF-8 biocomposite material, except that no enzyme is added.

[0060] The prepared acetylcholinesterase@ZIF-8 biocomposite was characterized by XRD, and the results were as follows Figure 4 As shown, the encapsulation of acetylcholinesterase did not affect the crystal structure of ZIF-8.

[0061] (2) Preparation of a...

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Abstract

The invention provides an acetylcholin esterase biosensor based on a metal organic framework as well as a preparation method and application of the acetylcholin esterase biosensor, and belongs to thefield of biological detection. The preparation method of the acetylcholin esterase biosensor comprises the following steps: mixing acetylcholin esterase and zinc acetate and oscillating, and then adding 2-methylimidazole solution for reaction to obtain acetylcholin esterase @ ZIF- 8 material; dissolving the acetylcholin esterase @ ZIF-8 material in water, mixing with a chloroauric acid solution, adding a sodium borohydride solution, and reacting to obtain acetylcholin esterase @ ZIF-8 @ Au nano ions; finally, mixing the acetylcholin esterase @ ZIF-8 material @ Au nano ion dispersion liquid with a chitosan solution, coating the surface of a glassy carbon electrode with the mixture, drying, and washing with water. The biosensor has the advantages of high sensitivity, strong stability, and good anti-interference and reproducibility, can be used for detecting organic pesticide residues, and has a detection limit as low as 10<-12>M.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a metal-organic framework-based acetylcholinesterase biosensor and its preparation method and application. Background technique [0002] Pesticides are widely used in modern agricultural production and people's daily life because of their high-efficiency insecticidal and herbicidal effects. However, due to the widespread use of pesticides, it will also lead to serious environmental pollution, making the pesticide residues in water, food and agricultural products exceed the standard, seriously affecting human food safety and health. Now the governments of various countries continue to strengthen the detection of pesticide residues, and the standards formulated are becoming more and more stringent. [0003] At present, the main methods for the analysis and detection of pesticide residues are gas chromatography, liquid chromatography, high performance liquid chromatography and v...

Claims

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Application Information

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IPC IPC(8): G01N27/327
CPCG01N27/327
Inventor 高发明赵奕淞卢雄杨云霞刘倩
Owner YANSHAN UNIV