Electrochemical biosensor based on polypeptide analog with electrocatalytic activity for acetylcholin esterase detection

An acetylcholinesterase and biosensor technology, applied in the field of functional materials and biosensing, to achieve the effects of high sensitivity detection, good biocompatibility and accurate results

Active Publication Date: 2018-06-05
NINGBO UNIV
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  • 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 molec

Method used

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  • Electrochemical biosensor based on polypeptide analog with electrocatalytic activity for acetylcholin esterase detection
  • Electrochemical biosensor based on polypeptide analog with electrocatalytic activity for acetylcholin esterase detection
  • Electrochemical biosensor based on polypeptide analog with electrocatalytic activity for acetylcholin esterase detection

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Example Embodiment

[0038] Example 1

[0039] The electrochemical biosensor based on peptide mimics with electrocatalytic activity is used for the detection of acetylcholinesterase, the specific steps are as follows:

[0040] (1) Synthesis of peptide mimics with electrocatalytic activity

[0041] Pipette 80.0μL of double distilled water and 10.0μL of silver nitrate (AgNO 3 ) Solution and 10.0μL L-Cysteine ​​(L-Cys) solution with a concentration of 1.0mM, stir vigorously on a stirrer for 3 minutes until the mixture is uniform, and then move to a shaker with a temperature of 37℃ and a rotation speed of 150r / min After incubating for 10 minutes, the cysteine-Ag(I) coordination polymer (abbreviated as Cys-Ag(I)CP) is obtained, which has a structure similar to a polypeptide. Finally, this peptide analog was moved to a refrigerator at 4°C and protected from light for later use.

[0042] (2) Preparation of electrochemical biosensor

[0043] a. First polish and clean the bare glassy carbon electrode (GCE, 3mm in ...

Example Embodiment

[0049] Example 2

[0050] The electrochemical biosensor based on peptide mimics with electrocatalytic activity is used for the detection of acetylcholinesterase, the specific steps are as follows:

[0051] (1) Synthesis of peptide mimics with electrocatalytic activity

[0052] Pipette 50.0μL of double-distilled water, 25.0μL of silver nitrate (AgNO 3 ) Solution and 25.0μL of 2.0mM L-Cys (L-Cys) solution, stir vigorously on a stirrer for 4min until the mixture is uniform, then move to a shaker with a temperature of 30℃ and a rotation speed of 160r / min After incubating for 18 min in medium, cysteine-Ag(I) coordination polymer (abbreviated as Cys-Ag(I)CP) is obtained, which has a structure similar to a polypeptide. Finally, this peptide analog was moved to a refrigerator at 4°C and protected from light for later use.

[0053] (2) Preparation of electrochemical biosensor

[0054] a. First polish and clean the bare glassy carbon electrode (GCE, 3mm in diameter), and dry it with nitrogen;

...

Example Embodiment

[0060] Example 3

[0061] The electrochemical biosensor based on peptide mimics with electrocatalytic activity is used for the detection of acetylcholinesterase, the specific steps are as follows:

[0062] (1) Synthesis of peptide mimics with electrocatalytic activity

[0063] Pipette 70.0μL of double distilled water, 15.0μL of 2.2mM silver nitrate (AgNO 3 ) Solution and 15.0μL L-Cysteine ​​(L-Cys) solution with a concentration of 2.2mM, stir vigorously on a stirrer for 2 minutes until the mixture is uniform, and then move to a shaker with a temperature of 29°C and a rotation speed of 190r / min After incubating for 10 minutes, the cysteine-Ag(I) coordination polymer (abbreviated as Cys-Ag(I)CP) is obtained, which has a structure similar to a polypeptide. Finally, this peptide analog was moved to a refrigerator at 4°C and protected from light for later use.

[0064] (2) Preparation of electrochemical biosensor

[0065] a. First polish and clean the bare glassy carbon electrode (GCE, 3mm...

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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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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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