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An electrochemical immunosensor based on Prussian blue and an electrochemical immunosensing method and application based on the sensor

An immunosensor, Prussian blue technology, applied in the field of electrochemical immunosensing and electrochemical immunosensor, can solve the problems of dissolved oxygen interference, complex operation, poor repeatability, etc. quick effect

Active Publication Date: 2021-01-26
HUBEI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to provide a Prussian blue-based electrochemical immunosensor and an electrochemical immunosensor based on the sensor based on the traditional method’s complex operation, poor repeatability, and usually facing problems such as dissolved oxygen interference. application, the sensor based on chitosan-Prussian blue (CS-PB) nanocomposite has the advantages of simple preparation process, controllability and good stability, and the urease functionalized silicon nanoprobe for signal transduction can be The catalytic reaction realizes multi-means electrochemical signal amplification, so that the method has excellent analytical performance such as high sensitivity, wide linear range, and low cost, and has good practical prospects in the field of clinical tumor marker detection

Method used

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  • An electrochemical immunosensor based on Prussian blue and an electrochemical immunosensing method and application based on the sensor
  • An electrochemical immunosensor based on Prussian blue and an electrochemical immunosensing method and application based on the sensor
  • An electrochemical immunosensor based on Prussian blue and an electrochemical immunosensing method and application based on the sensor

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Embodiment 1

[0029] A kind of electrochemical immunosensor based on Prussian blue of the present embodiment, described electrochemical immunosensor is that the base electrode is glassy carbon electrode, and its surface is modified chitosan-Prussian blue nanocomposite, gold nanoparticle and tumor mark in sequence Composition of object capture antibodies.

[0030] A kind of preparation method of the electrochemical immunosensor based on Prussian blue of the present embodiment, comprises the following steps:

[0031] (1) Preparation of chitosan-Prussian blue nanocomposite

[0032] Add 2 mL of 50 mM FeCl to 5 mL of 5 mg / mL chitosan (CS) with a mass fraction of 1% acetic acid 3 , 50mM K 3 Fe(CN) 6 and 0.1% acetic acid solution with a mass fraction of 1.0M KCl, mix well, heat up to 95°C, and react for 12 hours under stirring; the obtained product is centrifuged at 10,000rpm for 25min, the supernatant is discarded, and then washed with a 1% acetic acid solution Redisperse in 5.0mL 1% acetic a...

Embodiment 2

[0036] A kind of electrochemical immunosensing method based on the electrochemical immunosensor of Prussian blue of the present embodiment, comprises the following steps:

[0037] (1) Preparation of electrochemical immunosensor

[0038] Add 2 mL of 50 mM FeCl to 5 mL of 5 mg / mL chitosan (CS) with a mass fraction of 1% acetic acid 3 , 50mM K 3 Fe(CN) 6 and 0.1% acetic acid solution with a mass fraction of 1.0M KCl, mix well, heat up to 95°C, and react for 12 hours under stirring; the obtained product is centrifuged at 10,000rpm for 25min, the supernatant is discarded, and then washed with a 1% acetic acid solution Redisperse in 5.0mL of 1% acetic acid solution to obtain the CS-PB nanocomposite; add 5µL CS-PB nanocomposite to the glassy carbon electrode (GCE), drop it on the surface after drying at room temperature Add 8 µL of gold nanoparticles (Au-NP) with an average particle size of 13 nm and assemble at room temperature for 8 h; after washing with ultrapure water, continu...

Embodiment 3

[0041] Example 3 Detection of tumor marker CEA in standard solution

[0042] Add 8 µL of CEA standard solutions of different concentrations to the surface of the electrochemical immunosensor prepared in step (1) of Example 2, incubate and react at room temperature for 50 minutes, and then alternately wash with PBST solution and PBS buffer solution with pH = 6.9; continue to drop 8 µL of the urease-functionalized silicon nanoprobe dispersion prepared in step (2) of Example 2 was incubated at room temperature for 50 min, and then washed alternately with PBST solution and PBS buffer solution with pH = 6.9; then 8 µL containing 0.1 The aqueous solution of M urea (urea) and 0.1M dopamine (DA) was reacted at room temperature for 20min and then washed with ultrapure water; finally placed in 50mM pH=6.9PBS buffer solution containing 0.1M KCl, the Record the current response value in the range of 0.7-0V, thereby detecting the content of CEA in the standard solution, and the working cur...

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Abstract

The invention discloses a Prussian blue-based electrochemical immunosensor, an electrochemical immunosensing method built based on the sensor and application. The electrochemical immunosensor is composed of a glassy carbon electrode serving as a base electrode, and a glycan-Prussian blue nanocomposite, gold nanoparticles and a tumor marker capture antibody which are modified on the surface of thebase electrode in sequence. The electrochemical immunosensor is combined with a urease functionalized silicon nanoprobe to determine the content of a tumor marker in a sample. The Prussian blue-basedelectrochemical immunosensor has high analysis properties such as simple preparation process, controllability, high stability, high sensitivity, wide linear range and low cost, and has an extremely practical prospect in the field of clinical tumor marker detection.

Description

technical field [0001] The invention relates to the technical field of electrochemical biosensing, in particular to an electrochemical immunosensor based on Prussian blue and an electrochemical immunosensing method and application based on the sensor. Background technique [0002] Accurate determination of tumor markers is of great significance for early screening and clinical diagnosis of cancer. Compared with other methods, the immunosensor developed by combining highly specific immune recognition reaction with various optical and electrical sensing technologies has unique advantages such as high selectivity, small sample consumption, simple operation, and fast analysis speed. In recent years, it has been widely studied and applied in the detection of tumor markers. Among them, the electrochemical immunosensor developed based on the electroanalytical chemical method has a good practical prospect in the field of clinical diagnosis because it does not require expensive larg...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327G01N27/48
CPCG01N27/3277G01N27/3278G01N27/48
Inventor 赖国松郭鹏
Owner HUBEI NORMAL UNIV
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