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Preparation method of aflatoxin B1 immunoreaction electrode

A technology of aflatoxin and immune response, which is applied in the direction of material analysis, measuring devices, instruments, etc. by electromagnetic means, can solve the problem of poor mechanical stability of the gold particle layer, affecting the repeatability of measurement results, and difficulties in the distribution and size of gold nanoparticles. Control and other issues to achieve the effect of improving sensitivity, reducing analysis cost, improving specificity and utilization

Inactive Publication Date: 2015-06-24
HARBIN INST OF TECH AT WEIHAI
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  • Summary
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  • Application Information

AI Technical Summary

Problems solved by technology

However, the distribution and size of gold nanoparticles on the electrode are difficult to control, and the mechanical stability of the self-assembled gold particle layer is poor, which directly affects the repeatability of the measurement results.

Method used

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Embodiment 1

[0015] Embodiment 1: a kind of preparation method of aflatoxin B1 immunoreaction electrode, it comprises the following steps:

[0016] (1) Preparation of poly-o-phenylenediamine modified gold electrode: 50 mmol / L o-phenylenediamine was dissolved in 1.0 mol / L H 2 SO 4 , a gold electrode with a diameter of 3mm was used as the working electrode, a platinum electrode was used as the counter electrode, and a mercurous sulfate electrode was used as the reference electrode. Cyclic voltammetry was used. The electropolymerization potential range was -0.6 V~1.0 V, and the scan rate was 50 mV. / s, the number of polymerization circles is 30 circles, and the constant temperature is 25 ℃, and the poly-o-phenylenediamine-modified gold electrode is prepared.

[0017] (2) Fixation of glutaraldehyde: insert the above-mentioned modified gold electrode into phosphate buffer solution of 2.5% glutaraldehyde, keep the temperature at 25 ℃, and the reaction time is 60min. . The amino group of poly-...

Embodiment 2

[0021] Embodiment 2: a kind of preparation method of aflatoxin B1 immunoreaction electrode, it comprises the following steps:

[0022] (1) Preparation of poly-o-phenylenediamine modified gold electrode: 50 mmol / L o-phenylenediamine was dissolved in 1.0 mol / L H 2 SO 4 , a gold electrode with a diameter of 3 mm is used as the working electrode, a platinum electrode is used as the counter electrode, and a mercurous sulfate electrode is used as the reference electrode. For 30 cycles, at a constant temperature of 25 °C, a poly-o-phenylenediamine-modified gold electrode was obtained.

[0023] (2) Fixation of glutaraldehyde: insert the above-mentioned modified gold electrode into phosphate buffer solution of 2.5% glutaraldehyde, keep the temperature at 30 ℃, and the reaction time is 40min. Immobilization of glutaraldehyde on the basis of electropolymerized poly-o-phenylenediamine modified gold electrode. The amino group of poly-o-phenylenediamine reacts with the aldehyde group of ...

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Abstract

The invention relates to a preparation method of an aflatoxin B1 immunoreaction electrode. The preparation method comprises the following steps: preparing a poly-o-phenylenediamine modified gold electrode, and immobilizing glutaraldehyde on the basis of the electrically-polymerized poly-o-phenylenediamine modified gold electrode; and effectively immobilizing the protein A on the glutaraldehyde, facilitating the Schiff base generation reaction between an amino group of the protein A and the glutaraldehyde, wherein the immobilization quantity and firmness of the protein A on the surface of the electrode can be effectively improved. The AFB1 antibody immobilized on the protein A / glutaraldehyde / poly-o-phenylenediamine / gold electrode can maintain the spatial conformation, so that Fab fragments can orderly stretch out of the surface, the spatial resistance for the combination of the antibody and the antigen can be effectively reduced, the specificity and utilization rate of the antibody can be improved, and the sensitivity of the immunoreactions electrode can be improved. The preparation method is simple, the antibody is reliable and firm to immobilize, the specificity and the utilization rate of the antibody can be greatly improved, the operation is simple, the sensitivity is high, and rapidness in measurement can be realized.

Description

technical field [0001] The invention relates to a preparation method of an immune response electrode, in particular to a preparation method of an aflatoxin B1 immune response electrode. Background technique [0002] Aflatoxin (AF) is the most toxic class of biological toxins found so far, which can induce mutations, suppress immunity and cause cancer. of which aflatoxin B 1 (AFB 1 ) carcinogenicity, mutagenicity and teratogenicity all rank first. For this reason, many countries have imposed strict restrictions on its content in food and feed, and the European Union stipulates that AFB 1 The maximum detectable amount shall not exceed 2ng / g. Rapid and accurate analysis is an effective means to avoid and reduce the harm of AF. At present, the analysis and detection methods of AFB1 mainly include thin-layer chromatography, high performance liquid chromatography, enzyme-linked immunoassay, radioimmunoassay, gold standard method, immunoaffinity column purification-fluorescenc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327
Inventor 曹立新胡海峰王凯李宜
Owner HARBIN INST OF TECH AT WEIHAI
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