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Preparation method and application of immunosensor based on dopamine biomemetic modification

An immunosensor and dopamine technology, which is applied in the field of preparation of enzyme-free electrochemical immunosensor electrodes, can solve the problems of complex electrode modification steps and poor reproducibility, and achieve the effect of solving difficult synthesis, easy operation, and simple steps

Active Publication Date: 2014-02-12
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] One of the purposes of the present invention is to provide a simple and easy method for preparing an enzyme-free electrochemical immunosensor electrode to solve the problems of complex electrode modification steps and poor reproducibility

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] (1) Dopamine biomimetic modification: pipette 3 μL of freshly prepared 0.5 mg·mL -1 1. Dopamine hydrochloride solution with a pH of 8.5 was drip-coated on the surface of the treated and activated 3 mm glassy carbon electrode, and dried at room temperature;

[0024] (2) In situ preparation of nanoparticles: Rinse the electrode prepared in step (1) with ultrapure water several times, and then pipette 3 μL of 0.5 mg·mL -1 The silver nitrate solution is drip-coated on the surface of the electrode prepared in step (1), and dried at room temperature;

[0025] (3) Antibody immobilization: Rinse the electrode prepared in step (2) with ultrapure water several times, and apply the carcinoembryonic antigen (CEA) antibody solution on the surface of the electrode prepared in step (2) with a pipette gun, at 4°C dry under condition;

[0026] (4) Non-specific site blocking: Rinse the electrode prepared in step (3) with ultrapure water several times, and use a pipette gun to drop-coat...

Embodiment 2

[0030] (1) Biomimetic modification of dopamine: pipette 5 μL of freshly prepared 0.3 mg·mL -1 1. Dopamine hydrochloride solution with a pH of 8.5 was drip-coated on the surface of the treated and activated 4 mm glassy carbon electrode, and dried at room temperature;

[0031] (2) In situ preparation of nanoparticles: Rinse the electrode prepared in step (1) with ultrapure water several times, and then pipette 5 μL of 1.0 mg·mL -1 The chloroplatinic acid solution is drip-coated on the surface of the electrode prepared in step (1), and dried at room temperature;

[0032] (3) Antibody immobilization: Rinse the electrode prepared in step (2) with ultrapure water several times, and use a pipette gun to apply the prostate-specific antigen (PSA) antibody solution onto the surface of the electrode prepared in step (2), 4 Dry at ℃;

[0033] (4) Non-specific site blocking: Rinse the electrode prepared in step (3) with ultrapure water several times, and use a pipette gun to drop-coat th...

Embodiment 3

[0037] (1) Biomimetic modification of dopamine: Use a pipette to pipette 10 μL of the newly prepared concentration of 0.1 mg·mL -1 1. Dopamine hydrochloride solution with a pH of 8.5 is drip-coated on the surface of the treated and activated 5 mm gold electrode, and dried at room temperature;

[0038] (2) In situ preparation of nanoparticles: Rinse the electrode prepared in step (1) with ultrapure water several times, and then pipette 10 μL of 3.0 mg·mL -1 Palladium chloride solution is drip-coated on the electrode surface that step (1) makes, room temperature dries;

[0039] (3) Antibody immobilization: Rinse the electrode prepared in step (2) with ultrapure water several times, and apply the alpha-fetoprotein (AFP) antibody solution on the surface of the electrode prepared in step (2) with a pipette gun, at 4°C dry under condition;

[0040] (4) Non-specific site blocking: Rinse the electrode prepared in step (3) with ultrapure water several times, and use a pipette gun to ...

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Abstract

The invention discloses a preparation method and application of a non-enzymatic electrochemical immunosensor based on dopamine biomemetic modification and relates to the fields of nano-science, biological immune technology, electrochemical sensing and the like. An electrode is functionalized by utilizing dopamine biomemetic modification, and platinum, silver, palladium and other metal nanoparticles with enzymatic activity are synthesized in situ by utilizing the reduction characteristic of a poly-dopamine modification layer; an antibody is immobilized on the surface of the electrode by means of the interaction of the metal nanoparticles and poly-dopamine modification layer; electrochemical immunoassay on a tumor marker can be realized based on the electrocatalytic effect of the metal nanoparticles to an H2O2 reduction reaction and the identification effect of the antibody molecule to an antigen. The method is simple in steps, easy to operate and low in cost, has a high electrochemical activity and excellent response performance, can solve the problem of difficult nano-material synthesis in a conventional method and lack of simple and effective method, is applicable to preparation of multiple tumor marker immunosensor electrodes, and has a wide application prospect in scientific research and clinical application.

Description

technical field [0001] The invention relates to the fields of nanoscience, biological immune technology, electrochemical sensing and the like, and in particular to a preparation method and application of an enzyme-free electrochemical immunosensor electrode based on bionic modification of dopamine. Background technique [0002] Tumor marker detection is currently the only non-invasive method for early warning of cancer, which is of great significance in clinical screening, diagnosis and prognosis of tumors. Among the many tumor marker detection methods, electrochemical immunoassay has the advantages of simple operation, small sample volume, rapid sensitivity, low cost and easy miniaturization, etc., and has been widely used in clinical and scientific research. [0003] Early electrochemical immunosensors usually detect signals by enzymatically labeling antigens or antibodies, but the enzyme labeling operation is time-consuming, and the enzyme has high environmental requireme...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/531G01N27/416
CPCG01N33/5438G01N33/57407G01N33/9413G01N2800/7028
Inventor 马洪敏魏琴杨焱明高丕成张勇吴丹范大伟
Owner UNIV OF JINAN
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