Method, kit and application of copper oxide nanoparticle-labeled antibody

A nanoparticle and labeled antibody technology, applied in the field of immunoassay and diagnosis, can solve the problems affecting the detection limit of the test substance, environmental pollution and health, complex methods, etc., and achieve the effects of low cost, good stability and simple operation process.

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 above three methods to detect certain antigens or specific proteins, it is necessary to use various instruments in the readout method.
Among them, the fluorescein labeling technology needs a fluorescent microscope, the enzyme labeling technology needs a microplate reader, and the radionuclide technology needs automatic counters and other detection instruments. The dependence on instruments determines that many tests cannot be carried out in backward and underdeveloped areas.

Method used

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  • Method, kit and application of copper oxide nanoparticle-labeled antibody
  • Method, kit and application of copper oxide nanoparticle-labeled antibody
  • Method, kit and application of copper oxide nanoparticle-labeled antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Embodiment 1: the synthesis of compound 1

[0057] Compound 1

[0058] step:

[0059] 1. Vacuumize a 100mL two-neck flask (19#) and fill it with nitrogen.

[0060] 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.

[0061] 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.

[0062] 4. Remove the desiccant anhydrous sodium sulfate by filtration, distill the filtrate under reduced pressure, and concentrate to obtain ...

Embodiment 2

[0063] Embodiment 2: the synthesis of compound 2

[0064] Compound 2

[0065] step:

[0066] 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.

[0067] 2. The mixed solution was stirred at 5° C. for one hour, and then reacted at room temperature for 24 hours.

[0068] 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

[0069] Embodiment 3: the synthesis of compound 3

[0070]

[0071] step:

[0072] 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.

[0073] 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.

[0074] 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 an antibody labeling method, and especially relates to a method for labeling a copper oxide nano particle on an antibody through a simple physical absorption. The antibody labeling method of the invention has good universality; a reading mode of a detection needs no apparatus, and the reading can be finished by naked eyes; therefore, the method is suitable for field work. The invention also provides application of the antibody labeling method. The invention further provides a kit for antibody labeling and 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 labeling antibodies with nanoparticles and applications thereof. 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, in fluorescein labeling technology, fluorescein has the problems of fluorescence lifetime and fluorescence efficiency, and the method of labeling on ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/532G01N33/569
CPCC01P2002/85C01P2002/72G01N33/587B82Y30/00C01G3/02G01N33/532C01P2002/84G01N33/56983C01P2004/64
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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