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An electrochemical biosensor based on an electrocatalytically active peptide mimetic for the detection of acetylcholinesterase

A technology of acetylcholinesterase and biosensor, which is applied in the field of functional materials and biosensing to achieve the effects of accurate results, low cost and less reagent consumption

Active Publication Date: 2020-10-16
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no reports on electrochemical biosensors based on AChE and organophosphorus small molecule inhibitors based on electrocatalytically active polypeptide mimics modified electrodes at home and abroad.

Method used

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  • An electrochemical biosensor based on an electrocatalytically active peptide mimetic for the detection of acetylcholinesterase
  • An electrochemical biosensor based on an electrocatalytically active peptide mimetic for the detection of acetylcholinesterase
  • An electrochemical biosensor based on an electrocatalytically active peptide mimetic for the detection of acetylcholinesterase

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Experimental program
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Effect test

Embodiment 1

[0040] An electrochemical biosensor based on a polypeptide mimic with electrocatalytic activity is used for the detection of acetylcholinesterase, and the specific steps are as follows:

[0041] (1) Synthesis of peptide mimics with electrocatalytic activity

[0042] Pipette 80.0 μL double distilled water, 10.0 μL silver nitrate (AgNO 3 ) solution and 10.0 μL of L-cysteine ​​(L-Cys) solution with a concentration of 1.0 mM, stirred vigorously on a stirrer for 3 minutes until evenly mixed, and then moved to an oscillator with a temperature of 37 °C and a speed of 150 r / min Incubate in medium for 10 min to obtain a cysteine-Ag(I) coordination polymer (abbreviated as Cys-Ag(I)CP), which has a polypeptide-like structure. Finally, the peptide analogue was moved to a 4°C refrigerator and stored away from light for later use.

[0043] (2) Preparation of electrochemical biosensors

[0044] a. First, polish and clean the bare glassy carbon electrode (GCE, 3mm in diameter), and dry it ...

Embodiment 2

[0051] An electrochemical biosensor based on a polypeptide mimic with electrocatalytic activity is used for the detection of acetylcholinesterase, and the specific steps are as follows:

[0052] (1) Synthesis of peptide mimics with electrocatalytic activity

[0053] Pipette 50.0 μL double distilled water, 25.0 μL silver nitrate (AgNO 3 ) solution and 25.0 μL of L-cysteine ​​(L-Cys) solution with a concentration of 2.0 mM, stirred vigorously on a stirrer for 4 minutes until evenly mixed, and then moved to an oscillator with a temperature of 30 ° C and a speed of 160 r / min Incubate in medium for 18 min to obtain a cysteine-Ag(I) coordination polymer (abbreviated as Cys-Ag(I)CP), which has a structure similar to a polypeptide. Finally, the peptide analogue was moved to a 4°C refrigerator and stored away from light for later use.

[0054] (2) Preparation of electrochemical biosensors

[0055] a. First, polish and clean the bare glassy carbon electrode (GCE, 3mm in diameter), an...

Embodiment 3

[0062] An electrochemical biosensor based on a polypeptide mimic with electrocatalytic activity is used for the detection of acetylcholinesterase, and the specific steps are as follows:

[0063] (1) Synthesis of peptide mimics with electrocatalytic activity

[0064] Pipette 70.0 μL double distilled water, 15.0 μL silver nitrate (AgNO 3 ) solution and 15.0 μL of L-cysteine ​​(L-Cys) solution with a concentration of 2.2 mM, stirred vigorously on the stirrer for 2 minutes until uniformly mixed, and then moved to an oscillator with a temperature of 29 ° C and a speed of 190 r / min Incubate in medium for 10 min to obtain a cysteine-Ag(I) coordination polymer (abbreviated as Cys-Ag(I)CP), which has a polypeptide-like structure. Finally, the peptide analogue was moved to a 4°C refrigerator and stored away from light for later use.

[0065] (2) Preparation of electrochemical biosensors

[0066] a. First, polish and clean the bare glassy carbon electrode (GCE, 3mm in diameter), and d...

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Abstract

The invention discloses an electrochemical biosensor based on polypeptide analog with the electrocatalytic activity for acetylcholin esterase detection. The biosensor for acetylcholin esterase detection is characterized by comprising the following steps: (1) intensively stirring and uniformly mixing silver nitrate, L-cysteine and secondary distilled water on a stirrer, and incubating in an oscillator; (2) electrodepositing graphene onto a bare glassy carbon electrode by utilizing an electrochemical method, mixing 0.05 weight percent of a Nafion solution with the solution obtained in the step (1), and dropwise coating GO / GCE to obtain an electrochemical biosensor based on the polypeptide analog with electrocatalytic activity (CP / GO / GCE); (3) performing acetylcholin esterase activity detection, namely catalyzing acetylcholine in one step to generate H2O2, uniformly mixing acetylcholine, AchE and choline oxidase, and incubating to generate H2O2, and detecting the solution with the sensorprepared in the step (2) with good specificity, high sensitivity, high detection speed, accurate and reliable result; and (4) performing AChE inhibitor malathion detection, namely uniformly mixing acetylcholine, AChE, malathion and choline oxidase, incubating, and detecting the solution with the sensor prepared in the step (2). The biosensor has the advantages of good specificity, high sensitivity, high detection speed and accurate and reliable result.

Description

technical field [0001] The invention relates to an electrochemical biosensor, in particular to a preparation method of an electrochemical biosensor based on a polypeptide mimic with electrocatalytic activity and its application in the detection of acetylcholinesterase, and belongs to the technical field of functional materials and biosensing. Background technique [0002] Acetylcholinesterase (AChE) is a cholinesterase with carboxypeptidase and aminopeptidase activities, which can selectively catalyze the hydrolysis of the substrate acetylcholine. It is closely related to the development and maturation of cells, and can promote the development of neurons and regeneration of nerves. In the human body, AChE is mainly distributed in the nervous system tissue, which can quickly catalyze the hydrolysis of the neurotransmitter acetylcholine and lead to the termination of nerve impulse transmission, so that the normal physiological functions of the human body are guaranteed. Studi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/48G01N27/30G01N27/327G01N1/28G01N1/38
CPCG01N1/28G01N1/38G01N27/308G01N27/327G01N27/3271G01N27/48G01N2001/386
Inventor 胡宇芳张青青徐利华王娇饶家佳郭智勇王邃
Owner NINGBO UNIV
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