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Preparation method and application of a nitrogen-sulfur double-doped graphene oxide-labeled sandwich-type immunosensor

An immunosensor, double-doping technology, applied in the field of immunoassay and biosensing, can solve the problems of long detection time, complicated operation, radioactive pollution, etc., and achieve the effects of good catalytic performance, increased specific surface area, and excellent electrical conductivity.

Inactive Publication Date: 2020-08-11
SHANDONG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are many detection methods for cardiac troponin, such as enzyme-linked immunosorbent assay, immunofluorescence, etc., but these methods require large-scale equipment during the detection process, the operation is complicated, the detection takes a long time, and there are problems such as radioactive contamination. Therefore, it is very important to invent a cardiac troponin immunosensor with strong specificity, high sensitivity, fast detection speed and easy operation.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1 A method for preparing a nitrogen-sulfur double-doped graphene oxide-labeled sandwich-type immunosensor, comprising the following steps:

[0043] (1) A glassy carbon electrode with a diameter of 3.0mm is covered with Al 2 o 3 Grinding with polishing powder, cleaning with ultrapure water;

[0044] (2) Take 6.0 µL, 1.0 mg / mL gold nanorod functionalized polydopamine dispersion and apply it onto the electrode surface, dry it at room temperature, rinse the electrode surface with ultrapure water, and dry it;

[0045] (3) Add 6.0 µL, 8.0 µg / mL marker capture antibody Ab 1 Add dropwise to the surface of the electrode, rinse with ultrapure water, and dry in a refrigerator at 4°C;

[0046] (4) Continue to add 3.0 µL, 0.8 mg / mL bovine serum albumin BSA solution dropwise to the electrode surface, rinse the electrode surface with ultrapure water, and dry it in a refrigerator at 4 °C;

[0047] (5) Add 6.0 µL, 0.00005 ~ 200 ng / mL of a series of marker antigen Ag solut...

Embodiment 2

[0049] Embodiment 2 A method for preparing a nitrogen-sulfur double-doped graphene oxide-labeled sandwich-type immunosensor, comprising the following steps:

[0050] (1) A glassy carbon electrode with a diameter of 4.0 mm is covered with Al 2 o 3 Grinding with polishing powder, cleaning with ultrapure water;

[0051] (2) Take 6.0 µL, 2.0 mg / mL gold nanorod functionalized polydopamine dispersion and apply it onto the electrode surface, dry it at room temperature, rinse the electrode surface with ultrapure water, and dry it;

[0052] (3) Add 6.0 µL, 10 µg / mL marker capture antibody Ab 1 Add it dropwise to the surface of the electrode, rinse it with ultrapure water, and dry it in a refrigerator at 4 °C;

[0053] (4) Continue to add 3.0 µL, 1.0 mg / mL bovine serum albumin BSA solution dropwise to the electrode surface, rinse the electrode surface with ultrapure water, and dry it in a refrigerator at 4 °C;

[0054] (5) Add 6.0 µL, 0.00005 ~ 200 ng / mL of a series of marker antige...

Embodiment 3

[0056] Embodiment 3 A method for preparing a nitrogen-sulfur double-doped graphene oxide-labeled sandwich-type immunosensor, comprising the following steps:

[0057] (1) A glassy carbon electrode with a diameter of 5.0 mm is covered with Al 2 o 3 Grinding with polishing powder, cleaning with ultrapure water;

[0058] (2) Take 6.0 µL, 3.0 mg / mL gold nanorod functionalized polydopamine dispersion and apply it onto the electrode surface, dry it at room temperature, rinse the electrode surface with ultrapure water, and dry it;

[0059] (3) Add 6.0 µL, 12 µg / mL marker capture antibody Ab 1 Add it dropwise to the surface of the electrode, rinse it with ultrapure water, and dry it in a refrigerator at 4 °C;

[0060] (4) Continue to add 3.0 µL, 1.2 mg / mL bovine serum albumin BSA solution dropwise to the electrode surface, rinse the electrode surface with ultrapure water, and dry it in a refrigerator at 4 °C;

[0061] (5) Add 6.0 µL, 0.00005 ~ 200 ng / mL of a series of marker antige...

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Abstract

The invention belongs to the technical field of immune analysis and biosensing, and provides a preparation method and application of a nitrogen-sulfur double-doped graphene oxide-labeled sandwich-type immunosensor. In the present invention, nitrogen-sulfur double-doped graphene oxide functionalized with thionine and gold-platinum core-shell dendritic nanoparticles is used as a catalytic material, which is incubated with a detection antibody as a marker, and polydopamine functionalized with gold nanorods is used as a Electrode modification materials. The constructed sandwich electrochemical immunosensor has realized the quantitative detection of cardiac troponin cTnI and cTnT through differential pulse voltammetry, which has high sensitivity, strong specificity, low detection limit, and early detection of cardiac troponin It has important scientific significance and application value.

Description

technical field [0001] The invention belongs to the technical field of immune analysis and biosensing, and relates to a preparation method and application of a nitrogen-sulfur double-doped graphene oxide-labeled sandwich-type immune sensor. Specifically, nitrogen-sulfur double-doped graphene oxide functionalized with thionine and gold-platinum core-shell dendritic nanoparticles was used as the catalytic material to label the detection antibody, and polydopamine functionalized with gold nanorods was used as the electrode modification material. Pulse voltammetry achieves sensitive detection of cardiac troponin. Background technique [0002] Acute myocardial infarction (AMI), as the first killer that endangers human life and health, has the characteristics of acute onset and high mortality. Cardiac troponin is currently considered to be a better marker of myocardial damage, and it plays an important role in the early diagnosis and treatment of AMI. Therefore, a method with hi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327G01N27/30G01N27/48G01N33/68G01N33/532
Inventor 李月云吕慧李新进徐振陈磊
Owner SHANDONG UNIV OF TECH
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