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Biological sensor, preparation method, application in detection of acetylcholin esterase, and method for detecting organophosphorus pesticide

A technology of acetylcholinesterase and biosensors, applied in the field of biosensors, can solve complex, low detection sensitivity, time-consuming and labor-intensive problems

Active Publication Date: 2018-03-20
天之泰生物技术研究院(珠海)有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, certain disadvantages of conventional strategies still exist, such as low detection sensitivity, false positive effects, time and labor consuming, and require complex instruments, which obviously limit these applied methods.

Method used

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  • Biological sensor, preparation method, application in detection of acetylcholin esterase, and method for detecting organophosphorus pesticide
  • Biological sensor, preparation method, application in detection of acetylcholin esterase, and method for detecting organophosphorus pesticide
  • Biological sensor, preparation method, application in detection of acetylcholin esterase, and method for detecting organophosphorus pesticide

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Experimental program
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Embodiment 1

[0059] A method for preparing a biosensor, comprising the steps of:

[0060] Mix the acetylcholine buffer solution and the acetylcholinesterase buffer solution and incubate at 37°C for 10--15min, then add the mixed solution into the TMB solution containing hydrogen peroxide.

Embodiment 2

[0062] A biosensor prepared by the method described in the embodiment.

Embodiment 3

[0064] A kind of biosensor detects the application of acetylcholinesterase, and specific detection method is:

[0065] Mix acetylcholine buffer solution with different concentrations of acetylcholinesterase buffer solution respectively, incubate at 37°C for 15 minutes, add the mixed solution into TMB solution containing hydrogen peroxide, the final concentration of acetylcholine is 10mM / L, and the final concentration of acetylcholinesterase The concentrations were 0.2mU / mL, 0.4mU / mL, 0.6mU / mL, 0.8mU / mL, 1.0mU / mL, 1.2mU / mL and 1.4mU / mL, respectively. Use ultraviolet-visible spectrometers to measure the absorption wave, construct the linear relationship between the absorbance and the concentration of acetylcholinesterase, and linearize the absorbance at 652nm, and obtain the linear standard curve equation A=-0.10885C+0.1937, and the correlation coefficient R 2 =0.99729, and then realize the detection of acetylcholinesterase. The result is as Figure 6A , 6B .

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Abstract

The invention provides a biological sensor, a preparation method, application in detection of acetylcholin esterase, and a method for detecting an organophosphorus pesticide. The method is characterized in that acetyl choline has mimic enzyme to catalyze and oxidize TMB (3,3',5,5'-tetramethylbenzidine) to discolor, and can be decomposed and deactivated by the acetylcholin esterase, but the decomposing of hydrogen peroxide cannot be catalyzed, so that the TMB solution cannot generate color developing reaction; a linear relationship of absorbance and acetylcholin esterase is built, and the acetylcholin esterase is quantitatively detected; furthermore, the organophosphorus pesticide can effectively inhibit the activity of the acetylcholin esterase, the acetylcholin esterase is deactivated, the deactivated acetylcholin esterase cannot decompose the acetyl choline, and the TMB solution is promoted to generate the color developing reaction, thereby detecting the organophosphorus pesticide.

Description

technical field [0001] The invention belongs to the field of biosensors, and in particular relates to a biosensor, a preparation method, an application for detecting acetylcholinesterase, and a method for detecting organophosphorus pesticides. Background technique [0002] Acetylcholinesterase (AChE), as a biocatalyst enzyme, plays an important role in almost all metabolic processes, such as controlling related reactions and providing materials and energy required for cell proliferation and metabolism. Acetylcholinesterase (AChE), as the primary cholinesterase, is responsible for biological signal transmission in the nervous system. It can specifically regulate acetylcholine levels by hydrolyzing acetylcholine to choline, resulting in the termination of neurotransmission at cholinergic synapses. Importantly, many neurodegenerative diseases, including Parkinson's disease, Alzheimer's disease, and Huntington's disease, are closely related to impaired AChE. It has been demonst...

Claims

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Application Information

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IPC IPC(8): G01N21/78G01N21/31
CPCG01N21/31G01N21/78
Inventor 王广凤韩挺
Owner 天之泰生物技术研究院(珠海)有限公司
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