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Multiple-antigen synchronous detection method of quantum dot mark fluorescent immune

A technology of labeling fluorescence and simultaneous detection, which is applied in the direction of fluorescence/phosphorescence, biological testing, material inspection products, etc., can solve the problems of simultaneous quantitative detection of multiple antigens without reporting, and achieve the effect of easy promotion, simple method, and narrow emission peak

Inactive Publication Date: 2009-05-27
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method is based on the fluorescence quenching of quantum dots by carbon nanotubes, and there is no report on the simultaneous quantitative detection of multiple antigens

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] A CdSe quantum dot-labeled antibody with a fluorescent wavelength is selected to prepare a quantum dot probe to detect an antigen. For nanoparticles, taking carbon nanotubes as an example, to establish a fluorescent immunoassay method labeled with quantum dots, take the following steps:

[0021] 1. Preparation of nanoprobes for carbon nanotube-labeled antibodies: Add 100 mg of carbon nanotubes purchased on the market into 10 ml of 60% nitric acid solution for 30 minutes of sonication, then stir and reflux for 24 hours at 100 ° C, using a pore size of 0.22 Filtrate through a micron filter membrane, wash with water to remove nitric acid, and dry the filtered solid at 60° C. for 12 hours in vacuum to obtain carboxyl-modified carbon nanotubes. Take 2 milligrams of carboxy-modified carbon nanotubes, and link them with 1 milligram of monoclonal antibodies against breast cancer-associated antigen (BRCAA1) under the action of linker carbodiimide to prepare carbon nanotube-label...

Embodiment 2

[0025] Quantum dot-labeled antibodies that emit two kinds of fluorescence wavelengths are selected to detect two antigens. For nanoparticles, taking magnetic particles as an example, to establish a fluorescent immunoassay method labeled with quantum dots, take the following steps:

[0026] 1. Preparation of nanoprobes for magnetic particle-labeled antibodies: first, ferrous sulfate and ferric chloride were used to prepare nanometer ferric oxide (Fe 3 o 4 ) particles, and then use organic molecules to treat nano-Fe 3 o 4Particles have functional groups (carboxyl, amino, aldehyde, mercapto, etc.) on their surface. Here, magnetic particles with carboxyl groups on the surface (with a particle size of about 50 nanometers) are selected, and 1 mg of carboxyl-modified magnetic particles are mixed with 2 mg of monoclonal antibody against carcinoembryonic antigen (CEA) under the action of linker carbodiimide. Connect to prepare a magnetic particle-labeled anti-CEA monoclonal antibod...

Embodiment 3

[0030] Quantum dot labels that emit three fluorescent wavelengths are selected to detect three antigens. For nanoparticles, magnetic particles and carbon nanotubes are selected, and a fluorescent immunoassay method labeled with quantum dots is established.

[0031] Add 10 μL of alpha-fetoprotein standard or sample to be tested, 10 μL of carcinoembryonic antigen standard or sample to be tested, and 10 μL of breast cancer antigen (BRCAA1) standard or sample to be tested in the test tube. After mixing, add 100 μL by using The carbon nanotube-labeled anti-BRCAA1 monoclonal antibody prepared in Example 1, the mixture composed of 100 μL magnetic particle-labeled anti-CEA monoclonal antibody prepared in Example 2 and 100 μL magnetic particle-labeled anti-AFP monoclonal antibody were reacted at room temperature for 30 minutes, and then added 200 μL of the mixture composed of the CdSe quantum dot-labeled anti-BRCAA1 polyclonal antibody prepared in Example 1, 200 μL of the CdTe quantum ...

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Abstract

The invention relates to a quantum-dot-labeled immunofluorescence multi-antigen simultaneous detection method, belonging to the detection technical field. The method of the invention comprises the steps that the objects, i.e., the antibodies of the antigen to be detected are respectively connected to the quantum dot and the nano-particle like magnetic nano-particle by using the fluorescence characteristics of the quantum dot, like multiple-wavelength excitation, high-strength fluorescence emission, narrow emission peak, symmetrical peak shape, and stable luminescence; the antigen to be detected and an excessive amount of quantum-dot-labeled antibody are added; by immune reaction, the objects , i.e., the antibodies of the antigen to be detected, are jointed with the antibodies fixed on the quantum dot and the nano-particle like magnetic nano-particle to result in the joint of the quantum dot and the nano-particle like magnetic nano-particle, so as to form the composition of the antigen and the antibody, i.e., complex; the complex and the unreacted quantum-dot-labeled substance, i.e., free substance are separated, the intensity of fluorescence signal of the free substance is detected, and quantitative detection is carried out for the antigen to be detected. The invention has high sensitivity and wide detection range, and can simultaneously detect multiple antigens with simple operation and at a low cost.

Description

technical field [0001] The invention relates to a fluorescence immunoassay detection method in the technical field of detection, in particular to a quantum dot-labeled fluorescence immunoassay method for synchronous detection of multiple antigens. Background technique [0002] Quantum dots (semiconductor nanoparticles), also called artificial atoms, are aggregates composed of a certain number of actual atoms, semiconductor compounds with a size less than 10nm, and are ideal fluorescent indicators. Utilizing the ultra-fast optical nonlinear response of quantum dots, (room temperature) photoluminescence, and quantum dots can produce different colors and different wavelengths of emitted light according to different raw materials and different particle sizes of the same raw material, and multiple groups can be realized. score detection. [0003] Found through literature retrieval to prior art, in 2004 Goldman E R etc. published in "AnalyticalChemistry" (Vol.76, No.3, P684-688) ...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N21/64
Inventor 崔大祥潘碧峰高峰贺蓉
Owner SHANGHAI JIAO TONG UNIV
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