Prussian blue-based electrochemical immunosensor, electrochemical immunosensing method built based on sensor and application

An immune sensor, Prussian blue technology, applied in the field of electrochemical immune sensor, electrochemical immune sensor, can solve the problems of complex operation, poor repeatability, dissolved oxygen interference, etc., to overcome cumbersome operation, fast detection speed, sample consumption small effect

Active Publication Date: 2019-03-08
HUBEI NORMAL UNIV
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  • 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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  • Prussian blue-based electrochemical immunosensor, electrochemical immunosensing method built based on sensor and application
  • Prussian blue-based electrochemical immunosensor, electrochemical immunosensing method built based on sensor and application
  • Prussian blue-based electrochemical immunosensor, electrochemical immunosensing method built based on sensor and application

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Embodiment 1

[0029] An electrochemical immunosensor based on Prussian blue in this embodiment, the electrochemical immunosensor is composed of a glassy carbon electrode as a substrate electrode, and the surface of which is sequentially modified with chitosan-Prussian blue nanocomposites, gold nanoparticles and tumor markers It is composed of capture antibodies.

[0030] The method for preparing an electrochemical immunosensor based on Prussian blue of this embodiment includes the following steps:

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

[0032] To 5mL 5mg / mL chitosan (CS) with a mass fraction of 1% acetic acid solution was added 2mL containing 50mM FeCl 3 , 50mMK 3 Fe(CN) 6 The mass fraction of 1.0M KCl and 1.0M KCl is 0.1% acetic acid solution. After mixing evenly, the temperature is raised to 95°C and reacted for 12h under stirring; the resulting product is centrifuged at 10000rpm for 25min, the supernatant is discarded, and then washed with 1% acetic acid solution. Re...

Embodiment 2

[0036] The electrochemical immunosensing method based on the Prussian blue electrochemical immunosensor of this embodiment includes the following steps:

[0037] (1) Preparation of electrochemical immunosensor

[0038] To 5mL 5mg / mL chitosan (CS) with a mass fraction of 1% acetic acid solution was added 2mL containing 50mM FeCl 3 , 50mMK 3 Fe(CN) 6 The mass fraction of 1.0M KCl and 1.0M KCl is 0.1% acetic acid solution. After mixing evenly, the temperature is raised to 95°C and reacted for 12h under stirring; the resulting product is centrifuged at 10000rpm for 25min, the supernatant is discarded, and then washed with 1% acetic acid solution. Re-disperse in 5.0 mL of 1% acetic acid solution to obtain CS-PB nanocomposite; drop 5 µL of CS-PB nanocomposite onto the glassy carbon electrode (GCE), and then drop onto the surface after drying at room temperature Add 8μL of gold nanoparticles (Au-NP) with an average particle size of 13nm and assemble at room temperature for 8h; after washi...

Embodiment 3

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

[0042] Add 8 µL of CEA standard solution of different concentrations to the surface of the electrochemical immunosensor prepared in step (1) of Example 2, after incubating the reaction at room temperature for 50 minutes, wash with a pH=6.9 PBST solution and PBS buffer solution alternately; continue dripping 8μL of the urease-functionalized silicon nanoprobe dispersion prepared in step (2) of Example 2 was incubated at room temperature for 50 minutes and then washed again with pH=6.9 PBST solution and PBS buffer solution alternately; then 8μL containing 0.1 An aqueous solution of M urea (urea) and 0.1M dopamine (DA), react at room temperature for 20 minutes, and then wash with ultrapure water; finally, place it in a 50mM pH=6.9PBS buffer solution containing 0.1M KCl, and use differential pulse voltammetry Record the current response value in the range of 0.7-0V to detect the content of CEA in the standard soluti...

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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 established based on the sensor. Background technique [0002] The accurate determination of tumor markers is of great significance for early cancer screening and clinical diagnosis. 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 chemistry method does not require expensive large-scale instruments and has high sensitivity, so it has a good prac...

Claims

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Application Information

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Patent Type & Authority Applications(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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