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Preparation method and application of a three-antibody dual-channel immunosensor for simultaneous detection of human nuclear matrix protein and carcinoembryonic antigen

A technology of nuclear matrix protein and immunosensor, which is applied in the preparation and application field of three-antibody dual-channel simultaneous detection of human nuclear matrix protein and carcinoembryonic antigen immunosensor, can solve the problems that cannot explain the disease status, and achieve the promotion of electron transfer, Effect of good electron transport, high sensitivity and specific detection

Active Publication Date: 2017-04-19
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, sometimes the detection of only one tumor marker cannot explain the disease status in the human body, while the simultaneous detection of multiple markers can determine the disease status of the patient

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Preparation method and application of a three-antibody dual-channel immunosensor for simultaneous detection of human nuclear matrix protein and carcinoembryonic antigen

[0042] (1) with Al 2 o 3 Polish the glassy carbon electrode with a diameter of 4 mm, and clean it with ultrapure water; add 6 µL, 1.0 mg / mL graphene oxide@polyaniline@chitosan hydrochloride GO@PANI@CS-HCl dispersion solution dropwise to the surface of the electrode, and dry at room temperature to form a film;

[0043] (2) Add 6 µL of gold nano-sol with a mass fraction of 1% dropwise, and dry at room temperature;

[0044] (3) Add 6 µL and 1 µg / mL human immunoglobulin antibody Ab(I) standard solution dropwise, 3 µL bovine serum albumin BSA solution with a mass fraction of 1%, wash with ultrapure water, and keep at 4°C dry in the refrigerator;

[0045] (4) Add 6 µL, 1 µg / mL human immunoglobulin antigen Ag(I) standard solution dropwise, rinse with ultrapure water, and dry in a refrigerator at...

Embodiment 2

[0050] Example 2 Preparation method and application of a three-antibody dual-channel simultaneous detection of human nuclear matrix protein and carcinoembryonic antigen immunosensor

[0051] (1) with Al 2 o 3 Polish the glassy carbon electrode with a diameter of 4 mm, and clean it with ultrapure water; add 6 µL, 1.5 mg / mL graphene oxide@polyaniline@chitosan hydrochloride GO@PANI@CS-HCl dispersion solution dropwise to the surface of the electrode, and dry at room temperature to form a film;

[0052] (2) Add 6 µL of gold nano-sol with a mass fraction of 2% dropwise, and dry at room temperature;

[0053] (3) Add 6 µL, 3 µg / mL human immunoglobulin antibody Ab(I) standard solution dropwise, 3 µL bovine serum albumin BSA solution with a mass fraction of 3%, wash with ultrapure water, and keep at 4°C dry in the refrigerator;

[0054] (4) Add 6 µL, 3 µg / mL human immunoglobulin antigen Ag(I) standard solution dropwise, rinse with ultrapure water, and dry in a refrigerator at 4°C; ...

Embodiment 3

[0059] Example 3 Preparation method and application of a three-antibody dual-channel simultaneous detection of human nuclear matrix protein and carcinoembryonic antigen immunosensor

[0060] (1) with Al 2 o 3 Polish the glassy carbon electrode with a diameter of 4 mm, and clean it with ultrapure water; add 6 µL, 2.0 mg / mL graphene oxide@polyaniline@chitosan hydrochloride GO@PANI@CS-HCl dispersion solution dropwise to the surface of the electrode, and dry at room temperature to form a film;

[0061] (2) 6 µL of gold nano-sol with a mass fraction of 5% was added dropwise, and dried at room temperature;

[0062] (3) Add 6 µL, 5 µg / mL human immunoglobulin antibody Ab(I) standard solution dropwise, 3 µL bovine serum albumin BSA solution with a mass fraction of 5%, wash with ultrapure water, and keep at 4°C dry in the refrigerator;

[0063] (4) Add 6 µL, 5 µg / mL human immunoglobulin antigen Ag(I) standard solution dropwise, rinse with ultrapure water, and dry in a refrigerator a...

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Abstract

The invention relates to a preparation method and an application of an immunosensor capable of simultaneously detecting human nuclear matrix protein and carcino-embryonic antigen with three antibodies and double channels, and belongs to the technical field of building of novel sensors. On the basis of good specificity between the antigen and the antibodies, the detection ranges of the human nuclear matrix protein and the carcino-embryonic antigen are increased by the sensor by virtue of a sandwich structure of a human immunoglobulin immunization pair; the sensitivity of the sensor is significantly improved; the detection limit of the sensor is reduced; and the preparation method is of great significance in early diagnosis of bladder cancer.

Description

technical field [0001] The present invention designs a three-antibody dual-channel strategy electrochemical immunosensor, which uses gold hybrid sulfide graphene Au-S-GS as the connection carrier, and uses human immunoglobulin antibody as the skeleton to support the antibody to connect the human nuclear matrix protein antibody and carcinoembryonic antigen antibody, through the different oxidation peak potentials of toluidine blue and neutral red in the antibody incubation of the labeling layer, the simultaneous detection of human nuclear matrix protein and carcinoembryonic antigen belongs to the development of new biosensing technology and new Sensing methods build technology domains. Background technique [0002] Cancer is a general term for a large class of malignant tumors. In a normal human body, the content of tumor markers is lower than a certain concentration. During the development of tumors, the abnormal expression of tumor cells or certain substances produced by h...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/574G01N27/48
CPCG01N27/48G01N33/57473G01N33/68G01N2800/7028
Inventor 魏琴任祥王琦吴丹庞雪辉马洪敏范大伟孙旭
Owner UNIV OF JINAN
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