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A method for visually detecting antigen-antibody reaction and its application

A technology of antibody reaction and reaction, which is applied in the direction of analysis, measurement device, biological test, etc. through chemical reaction of materials, can solve the problems of limited development and application of instruments, and achieve the effect of low cost, high sensitivity and simple operation process

Active Publication Date: 2017-12-26
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

Although it is widely used in experimental research and clinical diagnosis, the existing enzyme-linked immunoassay method needs to use a microplate reader to read out the detection signal, and its dependence on the instrument limits its development in backward and underdeveloped areas. and application

Method used

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  • A method for visually detecting antigen-antibody reaction and its application
  • A method for visually detecting antigen-antibody reaction and its application
  • A method for visually detecting antigen-antibody reaction and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047]Synthesis of Surface Alkyne Functionalized Gold Nanoparticles

[0048] Utilize chloroauric acid reduction method to prepare nano-gold, the obtained nano-gold is centrifuged under a high-speed centrifuge, then collects the precipitate, and the precipitate is dispersed in a mixed solvent of water and tert-butanol (water: tert-butanol=2:1 ). Disperse 4mL of the above nano-gold solution in a mixed solvent of 16mL of water and tert-butanol (water: tert-butanol=2:1), and then add 50 μL of sodium hydroxide (0.5M) to adjust the pH to 9. Add 100 μL of terminal alkyne-functionalized thiol ligand (10 mM) and 30 μL of terminal thiol-terminated polyethylene glycol (200 mM) under stirring, and obtain surface alkyne-functionalized gold nanoparticles after ligand exchange for 24 hours. particle.

Embodiment 2

[0050] Synthesis of surface azide-functionalized gold nanoparticles

[0051] Utilize chloroauric acid reduction method to prepare nano-gold, the obtained nano-gold is centrifuged under a high-speed centrifuge, then collects the precipitate, and the precipitate is dispersed in a mixed solvent of water and tert-butanol (water: tert-butanol=2:1 ). Disperse 4mL of the above nano-gold solution in a mixed solvent of 16mL of water and tert-butanol (water: tert-butanol=2:1), and then add 50 μL of sodium hydroxide (0.5M) to adjust the pH to 9. 100 μL of terminal azido-functionalized thiol ligand (10 mM) and 30 μL of terminal thiol-terminated polyethylene glycol (200 mM) were added under stirring, and the surface azido-functionalized ligand was obtained after 24 hours of ligand exchange. Gold Nanoparticles.

Embodiment 3

[0053] Effects of Different Substances in the Reaction System on the Click Chemistry Reaction

[0054] The surface alkyne-functionalized gold nanoparticles and the surface azido-functionalized gold nanoparticles were mixed in a ratio of 1:1 for later use. Add divalent copper ions, divalent copper ions and ascorbic acid, divalent copper and phosphorylated ascorbic acid, alkaline phosphatase, divalent copper and phosphorylated ascorbic acid and alkaline phosphatase to the mixture of gold nanoparticles, and react for a period Observe and record the change of solution color after time, the results are shown in figure 2 .

[0055] from figure 2 It can be seen that when divalent copper ions, divalent copper ions, phosphorylated ascorbic acid, and alkaline phosphatase are added respectively, the solution is red; , the solution turned blue. This result indicates that alkaline phosphatase can catalyze the hydrolysis of phosphorylated ascorbic acid, dephosphorylate it to produce r...

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Abstract

The present invention relates to a method for visually detecting antigen-antibody reactions and its application. The method uses enzymes to catalyze substrate reactions to produce reducing products, which reduce divalent copper ions to monovalent copper ions. The click chemical reaction catalyzed by gold nanoparticles and monovalent copper is used to visually analyze the enzyme content; combined with the enzyme-labeled antibody technology, the enzyme-labeled antibody is used in the immunoassay of antigen-antibody interaction, and the enzyme content is analyzed visually The method achieves the purpose of visual analysis of antigen-antibody interaction; the method is used for visual detection of relevant antigen-antibody in clinical samples. The invention can complete the detection of the antigen-antibody reaction only by naked eyes, and has the advantages of high sensitivity, simple operation, low cost, good stability and the like.

Description

technical field [0001] The invention belongs to the technical field of immune detection and analysis. It specifically relates to a method for detecting antigen-antibody reaction and its application, in particular to a method for visually detecting antigen-antibody reaction and its application. Background technique [0002] The immunolabeling analysis method is a biological analysis method developed by combining the high specificity of the immune response and the high sensitivity of the biomarker. This analytical method not only has high sensitivity, strong specificity, good repeatability and high accuracy, but also is simple to operate, easy to be commercialized and automated. [0003] The application range of immunolabeling analysis method is extremely wide. It can measure various biologically active substances such as endocrine hormones, proteins, nucleic acids, cell surface antigens, and tumor markers, which has played a role in promoting the development of basic medical...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/76G01N33/53G01N33/569
Inventor 蒋兴宇鲜于运雷
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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