Method, kit and application for visual detection of antigen-antibody reaction

A technology of antigen antibody and kit, which is applied in the field of surface-functionalized gold nanoparticles, can solve problems affecting the detection limit of analytes, environmental pollution and health, and easy inactivation of enzymes, achieving low cost, good stability, The effect of detection system stability

Active Publication Date: 2011-12-07
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, in fluorescein labeling technology, fluorescein has the problems of fluorescence lifetime and fluorescence efficiency, and the method of labeling on antigen or antibody is complicated; in enzyme labeling technology, the enzyme is easily inactivated, thus affecting the detection limit of the analyte; In the radionuclide labeling technology, nuclides are radioactive, and there are strong environmental pollution and health hazards
In addition, when using the ab

Method used

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  • Method, kit and application for visual detection of antigen-antibody reaction
  • Method, kit and application for visual detection of antigen-antibody reaction
  • Method, kit and application for visual detection of antigen-antibody reaction

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Embodiment 1: the synthesis of compound 1

[0067] Compound 1

[0068] step:

[0069] 1. Vacuumize a 100mL two-neck flask (19#) and fill it with nitrogen.

[0070] 2. Add 50 mL of toluene to the flask under nitrogen protection, and dissolve triphenylchloromethane (4.60 g, 16.5 mmol) and N, N-diisopropylethylamine (DIEA, 4.20 g, 33.0 mmol) in the toluene Inside, 11-mercaptoundecanoic acid (3.00 g, 13.8 mmol) was added under magnetic stirring, and reacted at room temperature for 5 hours under nitrogen protection.

[0071] 3. After the reaction is complete, distill the solvent off under reduced pressure, add 50 mL of dichloromethane to the residual product, and after fully dissolving, wash with saturated brine (3×100 mL) three times, then dry the dichloromethane solution with anhydrous sodium sulfate , let stand overnight.

[0072] 4. Remove the desiccant anhydrous sodium sulfate by filtration, distill the filtrate under reduced pressure, and concentrate to obtain ...

Embodiment 2

[0073] Embodiment 2: the synthesis of compound 2

[0074] Compound 2

[0075] step:

[0076] 1. Add compound 1 (1.50g, 3.3mmol) in a 50mL single-necked flask (19#), EDC-HCl (0.69g, 3.6mmol) catalytic amount of DMAP, add 25mL of anhydrous dichloromethane, magnetically stir to make After dissolution, N-hydroxysuccinimide (NHS, 0.45 g, 3.9 mmol) was finally added to the mixture.

[0077] 2. The mixed solution was stirred at 5° C. for one hour, and then reacted at room temperature for 24 hours.

[0078] 3. After the reaction, the reaction solution was diluted with 25 mL of dichloromethane, washed three times with saturated brine (3×50 mL), dried the organic phase with anhydrous sodium sulfate, and concentrated to obtain compound 2 (1.80 g, 3.23 mmol). is the activated ester of compound 1. The yield was 99%.

Embodiment 3

[0079] Embodiment 3: the synthesis of compound 3

[0080]

[0081] step:

[0082] 1. Add NH 2 C 2 h 4 OC 2 h 4 OC 2 h 4 NH 2 (8mL, 55mmol) was dissolved in anhydrous dichloromethane, and slowly added dropwise to the mixed solution 10mL of compound 2 (1.80g, 3.23mmol) in dichloromethane under stirring, after the addition was complete, react at room temperature overnight.

[0083] 2. Dilute the reaction solution with 50 mL of dichloromethane, wash the organic phase with saturated brine (3×50 mL) three times, dry the organic phase with anhydrous sodium sulfate, and concentrate under reduced pressure to obtain the crude compound 3.

[0084] 3. Dissolve the crude product with a little dichloromethane and apply it as a sample, then separate by column chromatography, the eluent is chloroform: methanol: ammonia water = 20:1:0.05, the pure product is a colorless oily liquid, a total of 1.78g (3.0mmol ), the yield was 94%.

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Abstract

The invention relates to a method for visual detection of an antigen-antibody reaction. The method is specifically characterized in that: copper oxide nanoparticle-labeled antibodies react with a sample requiring detection; surface functionalized gold nanoparticles are adopted to detect the copper oxide nanoparticles labeled on the antibodies so as to provide information related to the antigen-antibody reaction, and provide a new tool for the disease detection and the disease diagnosis based on the immunological reaction. With the method provided by the present invention, the detection can be completed by the naked eye without requiring any equipment, and the method is applicable for field work. The invention further provides an application of the method. The invention further provides a kit for the visual detection of the antigen-antibody reaction, and an application thereof.

Description

technical field [0001] The invention belongs to the technical field of immune analysis and diagnosis. Specifically, the present invention relates to a method for amplifying the signal of antibody labeling, especially by using a surface-functionalized gold nanoparticle, which can directly judge the antigen-antibody reaction with the naked eye by detecting the antibody label. Background technique [0002] When detecting certain antigens or specific proteins, immunolabeling techniques are usually applied. Immunolabeling technology is to mark some substances that are both easy to measure and highly sensitive to specific antigens or antibody molecules, and to display the nature and content of antigens or antibodies in the reaction system through the enhanced amplification of these markers. Currently commonly used labels include fluorescein, enzymes, and radionuclides. However, in practical applications, these three immunolabeling techniques have different defects. For example,...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/558G01N33/569
CPCG01N33/587
Inventor 蒋兴宇曲伟思刘颖昳王卓
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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