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Preparation method of oncofetal antigen electrochemical immunosensor based on AuNPs-PDDA-GR composite material and application thereof

A technology of immunosensor and carcinoembryonic antigen, applied in the field of electrochemical immunosensing

Inactive Publication Date: 2015-12-16
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the main challenge in the field of disease diagnosis is how to develop miniaturized, integrated and portable monitoring and analysis methods and equipment, and to establish a set of simple, fast, high-sensitivity methods that can be widely applied.

Method used

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  • Preparation method of oncofetal antigen electrochemical immunosensor based on AuNPs-PDDA-GR composite material and application thereof
  • Preparation method of oncofetal antigen electrochemical immunosensor based on AuNPs-PDDA-GR composite material and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] (2) Take 50 μL of 1.0 μg / mL carcinoembryonic antigen antibody (anti-CEA) standard solution and apply it dropwise on the surface of the electrode, incubate in a refrigerator at 4°C for 12 hours, wash with PBS (pH7.4) buffer solution to remove physical adsorption and obtain anti -CEA / AuNPs-PDDA-GR / GCE;

[0033] (3) Take 50 μL of bovine serum albumin (BSA) solution with a mass fraction of 1% drop-coated on the surface of the electrode, incubate at 37°C for 1 h to block the non-specific binding sites, and wash with PBS (pH 7.4) buffer solution BSA / anti-CEA / AuNPs-PDDA-GR / GCE was obtained on the electrode surface;

[0034] (4) Drop 50 μL of a series of carcinoembryonic antigen standard solutions with a concentration of 0.1pg / mL~10ng / mL for specific recognition with antibodies, incubate at 37°C for 60min, and buffer with PBS (pH7.4) The electrode surface was rinsed with the solution to prepare a carcinoembryonic antigen electrochemical immunosensor (CEA / BSA / anti-CEA / AuNPs-PDD...

Embodiment 2

[0037] (2) Take 50 μL of 2.0 μg / mL anti-CEA standard solution and drop-coat it on the surface of the electrode, incubate in a refrigerator at 4°C for 12 hours, wash with PBS (pH7.4) buffer solution to remove physical adsorption to obtain anti-CEA / AuNPs-PDDA -GR / GCE;

[0038] (3) 50 μL of BSA solution with a mass fraction of 2% was drip-coated on the electrode surface, incubated at 37°C for 1 h to block non-specific binding sites, and washed the electrode surface with PBS (pH 7.4) buffer solution to obtain BSA / anti -CEA / AuNPs-PDDA-GR / GCE;

[0039] (4) Drop 50 μL of a series of carcinoembryonic antigen standard solutions with a concentration of 0.1pg / mL~10ng / mL for specific recognition with antibodies, incubate at 37°C for 60min, and buffer with PBS (pH7.4) The electrode surface was rinsed with the solution to prepare a carcinoembryonic antigen electrochemical immunosensor (CEA / BSA / anti-CEA / AuNPs-PDDA-GR / GCE) based on AuNPs-PDDA-GR composite material.

[0040] Example 3 A meth...

Embodiment 3

[0042] (2) Take 50 μL of 2.5 μg / mL anti-CEA standard solution and drop-coat it on the surface of the electrode, incubate in a refrigerator at 4°C for 12 hours, wash with PBS (pH7.4) buffer solution to remove physical adsorption to obtain anti-CEA / AuNPs-PDDA -GR / GCE;

[0043] (3) Take 50 μL of BSA solution with a mass fraction of 3% and drop-coat it on the electrode surface, incubate at 37°C for 1 h to block the non-specific binding sites, wash the electrode surface with PBS (pH 7.4) buffer solution to obtain BSA / anti -CEA / AuNPs-PDDA-GR / GCE;

[0044] (4) Drop 50 μL of a series of carcinoembryonic antigen standard solutions with a concentration of 0.1pg / mL~10ng / mL for specific recognition with antibodies, incubate at 37°C for 60min, and buffer with PBS (pH7.4) The electrode surface was rinsed with the solution to prepare a carcinoembryonic antigen electrochemical immunosensor (CEA / BSA / anti-CEA / AuNPs-PDDA-GR / GCE) based on AuNPs-PDDA-GR composite material.

[0045] Example 4 Pre...

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Abstract

The invention relates to a preparation method of an oncofetal antigen electrochemical immunosensor based on AuNPs-PDDA-GR composite material and an application thereof, and belongs to the field of electrochemical immunization sensing technology. K3Fe(CN)6 / K4Fe(CN)6 is used as an electrochemical probe, the AuNPs-PDDA-GR composite material is used as a substrate material with good biological compatibility and stability, large specific surface area, excellent conductivity and other advantages, thereby substantially improving stability and sensitivity of the immunosensor. The sensor is used for detection of oncofetal antigens, and the detection linear scope is 0.1pg / mL-10ng / mL, and the detection limit is 0.05pg / mL.

Description

technical field [0001] The invention relates to the technical field of electrochemical immunosensing, in particular to a preparation method and application of a carcinoembryonic antigen electrochemical immunosensor based on AuNPs-PDDA-GR composite material. Specifically, an electrochemical immunosensor for detecting carcinoembryonic antigen was prepared by using AuNPs-PDDA-GR composite material as the base material. Background technique [0002] Carcinoembryonic antigen (CEA) is a broad-spectrum tumor marker that can reflect the existence of a variety of tumors. It plays a role as a functional colon cancer ligand. It is a better tumor marker in terms of control and development, and monitoring recurrence. The current immunological methods for the determination of CEA mainly include enzyme-linked immunoassay, fluorescent immunoassay, radioimmunoassay and so on. These methods have the advantage of being sensitive and reliable. However, there are certain defects, such as the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/574G01N33/543
CPCG01N33/68G01N33/5438G01N33/57473G01N2333/47
Inventor 郑香丽周长利田栋夏方诠花小霞
Owner UNIV OF JINAN
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