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Preparation method of electrochemical sensor of zearalenone specific polypeptide

A zearalenone and specific technology, applied in scientific instruments, instruments, material analysis through electromagnetic means, etc., to achieve the effect of increasing sensitivity, high sensitivity and accuracy, and increasing stability

Pending Publication Date: 2022-04-12
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, TLC, HPLC, GC and MS methods require expensive instruments and skilled operation skills. Therefore, in order to simplify the detection process, shorten the detection time and save the detection cost, people shift the focus of research to electrochemical sensors. Disposable carbon mesh Printed electrodes have the advantages of easy portability, timely inspection, fast response, easy operation, strong specificity, and strong anti-interference ability. At present, there is no method for combining peptides with disposable carbon screen-printed electrodes for the detection of zearalenone.

Method used

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  • Preparation method of electrochemical sensor of zearalenone specific polypeptide
  • Preparation method of electrochemical sensor of zearalenone specific polypeptide
  • Preparation method of electrochemical sensor of zearalenone specific polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Screening for specific polypeptides

[0041] Construct a polypeptide sequence library that specifically binds to zearalenone, establish a polypeptide sequence library through molecular simulation technology, and preliminarily screen out four polypeptide chains. The specific steps are as follows:

[0042] (1) Parent molecular docking: use Discovery Studio 2016 (DS 2016) molecular simulation software to preprocess the zearalenone receptor downloaded from the PDB protein database and the ligand downloaded from the chemical database, and set the receptor to 6JR9 :6JR9, the input ligand is ZEN:All, set the conformational cluster radius to 0.5, and set the other parameters according to the default state, apply the CHARMm force field, and click CDOCKER to run to dock the flexible molecules;

[0043] (2) Analysis of the docking results: Open the above successful docking result report, view its 2D action map, analyze its main role in the amino acid sequence, and determ...

Embodiment 2

[0048] Example 2 Screening the optimal specific polypeptide chain

[0049] The optimal specific polypeptide chain is screened by electrochemical methods: after the surface of the gold electrode is modified step by step, in the presence of redox probes, the electron transfer ability is inhibited, the peak current value gradually decreases, and the resistance gradually increases. In this invention, electrochemical AC impedance method was used to investigate the specific binding ability of the four polypeptide chains (L0-L3) screened in Example 1 to zearalenone by the change of resistance value. The specific operation steps are as follows:

[0050] (1) Pretreatment of the gold electrode: the gold electrode with a diameter of 3 mm was polished with alumina powder with a particle size of 0.3 μm and 0.05 μm in sequence, and then soaked in the piranha solution for 10 min, and after taking it out, use 50 %HNO 3 Ultrasonic cleaning with aqueous solution, absolute ethanol, and ultrapure...

Embodiment 3

[0055] Embodiment 3 prepares electrochemical sensor

[0056] A method for preparing an electrochemical sensor for a zearalenone-specific polypeptide, comprising the steps of:

[0057] (1) Pretreatment of carbon screen printing electrodes: before use, rinse the electrode surface with ultrapure water for 10s, and the treatment methods and methods of each electrode should be consistent, N 2 After drying, place in 5mmol / L K 3 [Fe(CN) 6 ] / K 4 [Fe(CN) 6 ] (containing 1mol / LKCl), set the potential range to -0.2-0.6V, scan speed to 100mV / s, and perform cyclic voltammetry scan. When a repeatable voltammogram appears, the activation is successful and can be carried out. In the next step of electrode modification, the processed electrode N 2 Blow dry and avoid light and dust for later use;

[0058] (2) Self-assembly of rGO-COOH-P-Atp / SPCE: Take 0.5mg / mL carboxylated graphene dispersion, add 0.25mg p-aminothiophenol, ultrasonicate at room temperature for 0.5h, mix the The suspensio...

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Abstract

The invention provides a preparation method of an electrochemical sensor of zearalenone specific polypeptide. The preparation method comprises the following steps: (1) pretreating a carbon screen-printed electrode; (2) modification of a carboxylated graphene and p-aminothiophenol composite material; (3) self-assembly of the nanogold material; (4) self-assembling by taking the specific polypeptide L2 as a recognition element; (5) closing an Au site which is not combined with the polypeptide L2 by using mercaptohexanol; and (6) detecting a reduction peak current value by using a differential pulse voltammetry method. According to the preparation method of the electrochemical sensor of the zearalenone specific polypeptide, a carboxylated graphene composite material of a p-aminothiophenol cross-linking agent is fixed on the surface of a carbon screen-printed electrode, so that the sensitivity is improved, and meanwhile, a larger specific surface area is provided for the next step of electro-deposition of gold nanoparticles.

Description

technical field [0001] The invention belongs to the field of food safety detection, and in particular relates to a preparation method of an electrochemical sensor for a zearalenone-specific polypeptide. Background technique [0002] Zearalenone (ZEN), an estrogen produced by Fusarium, is one of the most common non-steroidal mycotoxins in crops. This mycotoxin mainly accumulates before harvest. It also accumulates under poor storage conditions, so grains and their products are susceptible to mycotoxin contamination. ZEN has heat-resistant properties and will not be degraded by processing such as grinding, extrusion, storage or heating. Mycotoxin contamination in corn, wheat and other foods will cause economic losses and diseases, and bring risks to animals and humans. Toxic constituents may also be present in target organisms and in meat and dairy products. The European Commission regulations stipulate that the maximum limit in cereals, cereal flour and bran is 75 μg / kg, an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327
Inventor 刘冰彭嘉璇王硕
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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