Preparation method of gold nanorod-coupled horse radish peroxidase and carcino-embryonic antigen-labeled antibody

A technology of coupling horseradish peroxidase and gold nanorods, which is applied in measuring devices, instruments, scientific instruments, etc., can solve problems such as low sensitivity, low specificity, and insignificant early diagnosis of tumors, and achieves Solve the cumbersome steps and improve the effect of sensitivity

Inactive Publication Date: 2016-12-14
TIANJIN UNIV
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  • Abstract
  • Description
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Problems solved by technology

Carcinoembryonic antigen is a broad-spectrum tumor marker, which can reflect the existence of a variety of tumors to people. It is a good tumor marker for the judgment of curative effect, disease development, monitoring and prognosis estimation of colorectal cancer, breast cancer and lung cancer However, its specificity is not strong, its sensitivity is not high, and its role in the early diagnosis of tumors is not obvious

Method used

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  • Preparation method of gold nanorod-coupled horse radish peroxidase and carcino-embryonic antigen-labeled antibody
  • Preparation method of gold nanorod-coupled horse radish peroxidase and carcino-embryonic antigen-labeled antibody
  • Preparation method of gold nanorod-coupled horse radish peroxidase and carcino-embryonic antigen-labeled antibody

Examples

Experimental program
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Effect test

Embodiment example 1

[0027] (1) The molar mass ratio of 2.5 μmol is chloroauric acid (HAuCl 4 ) and 1000 μmol hexadecyl trimethyl ammonium bromide (Hexadecyl trimethyl ammonium Bromide CTAB) were added to the reactor, and 6 μmol sodium borohydride (NaBH) was added to the reactor. 4 ) to obtain gold seed solution;

[0028] (2) get above-mentioned gold seed solution and join into reactor, add molar mass ratio to be chloroauric acid: cetyl trimethyl ammonium bromide: ascorbic acid: silver nitrate=1:400:2:40, prepare gold Nano stave;

[0029] (3) 5 mg of gold nanorods (AuNRs) were added to the reactor, the pH of the gold nanorods was adjusted to 9.0 by potassium carbonate aqueous solution, and 0.01 mg of carcinoembryonic antigen CEA-labeled antibody (Ab 1 ) and 0.04 mg of horseradish peroxidase (HRP) mixture were mixed with gold nanorods to obtain AuNRs, HRP and Ab 1 The mixed solution is rotated and mixed on a rotating mixing rack and then left to stand for treatment;

[0030] (4) Add 1 mg of bov...

Embodiment example 2

[0033] (1) The molar mass ratio of 2.5 μmol is chloroauric acid (HAuCl 4 ) and 1000 μmol hexadecyl trimethyl ammonium bromide (Hexadecyl trimethyl ammonium Bromide CTAB) were added to the reactor, and 6 μmol sodium borohydride (NaBH) was added to the reactor. 4 ) to obtain gold seed solution;

[0034] (2) get above-mentioned gold seed solution and join into reactor, add molar mass ratio to be chloroauric acid: cetyl trimethyl ammonium bromide: ascorbic acid: silver nitrate=1:400:2:80, prepare gold Nano stave;

[0035] (3) 5 mg of gold nanorods (AuNRs) were added to the reactor, the pH of the gold nanorods was adjusted to 9.0 by potassium carbonate aqueous solution, and 0.01 mg of carcinoembryonic antigen CEA-labeled antibody (Ab 1) and 0.04mg horseradish peroxidase (HRP) mixture mixed with gold nanorods, the resulting AuNRs, HRP and Ab 1 The mixed solution was rotated and mixed on the rotating mixing rack and then left to stand for processing;

[0036] (4) Add 1 mg bovine ...

Embodiment example 3

[0038] (1) 2.5 μmol molar mass ratio is chloroauric acid (HAuCl 4 ) and 1000 μmol cetyl trimethyl ammonium bromide (Hexadecyl trimethyl ammonium Bromide CTAB) were added to the reactor, and 6 μmol sodium borohydride (NaBH 4 ) to obtain gold seed solution;

[0039] (2) Get the above-mentioned gold seed solution and join the reactor, adding the molar mass ratio is chloroauric acid: hexadecyltrimethylammonium bromide: ascorbic acid: silver nitrate=1:400:2:120, and gold is prepared Nano stave;

[0040] (3) Add 5 mg of gold nanorods (AuNRs) into the reactor, adjust the pH of the gold nanorods to 9.0 by potassium carbonate aqueous solution, and add 0.01 mg of carcinoembryonic antigen CEA-labeled antibody (Ab 1 ) and 0.04mg horseradish peroxidase (HRP) mixture mixed with gold nanorods, the resulting AuNRs, HRP and Ab 1 The mixed solution was rotated and mixed on the rotating mixing rack and then left to stand for processing;

[0041] (4) Add 1 mg bovine serum albumin to the above...

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Abstract

The invention relates to a preparation method of a gold nanorod-coupled horse radish peroxidase and carcino-embryonic antigen-labeled antibody. A multienzyme-labeled gold nanoparticle alpha-fetoprotein immunoprobe is prepared through the electrostatic adsorption effect of a gold nanorod, a carcino-embryonic antigen-labeled antibody and horse radish peroxidase to conduct qualitative and quantitative detection on a carcino-embryonic antigen (CEA) which is a protein marker widely existing in a digestive system cancer of an endoblast source, a novel protein marker detection method is established, and sensitivity and specificity of detection are improved. The preparation method has the advantages that the whole preparation process is simple, and the method is suitable for industrialized production; the carcino-embryonic antigen can be qualitatively and quantitatively detected, and the specificity is good; the whole detection process is low in cost, easy and convenient to operate and suitable for community tumor screening of a high risk group, and a novel tumor detection method is established.

Description

technical field [0001] The invention relates to the technical field of nanomaterial preparation, and more particularly to a preparation method of a gold nanorod coupled with horseradish peroxidase and a carcinoembryonic antigen-labeled antibody. Background technique [0002] Enzyme is an organic catalyst. A small amount of enzyme can lead to a large number of catalytic processes. Immunoenzyme technology is a new technology established by combining the immune reaction of antigens and antibodies with the catalytic reaction of enzymes. After the enzyme is combined with the antibody or antigen, it neither changes the specificity of the immunological reaction of the antibody to the antigen nor affects the enzymatic activity of the enzyme itself, that is, with the participation of the corresponding and suitable substrate, the matrix is ​​hydrolyzed and colored. , or change the hydrogen donor from a colorless reduced form to a colored oxidized form. The colored reaction shows the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N33/574
CPCG01N33/535G01N33/57473
Inventor 常津武玉东宫晓群
Owner TIANJIN UNIV
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