Method for visually detecting antigen-antibody reaction and application of method

An antibody reaction and antigen technology, which is applied in the direction of chemical reaction of materials for analysis, measurement devices, biological tests, etc., can solve the problems of limited development and application of instruments, and achieve the effects of low cost, good stability and high sensitivity

Active Publication Date: 2014-10-08
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 rea

Method used

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  • Method for visually detecting antigen-antibody reaction and application of method
  • Method for visually detecting antigen-antibody reaction and application of method
  • Method for visually detecting antigen-antibody reaction and application of method

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 invention relates to a method for visually detecting antigen-antibody reaction and an application of the method. The method is characterized in that bivalent copper ions are reduced to monovalent copper ions by using a reducing product generated by reaction of enzyme catalysis substrates; the content of enzyme is visually analyzed through gold nanoparticles with the functionalized surfaces and a monovalent-copper-catalyzed click chemical reaction; an enzyme-labeled antibody is applied to immunoassay with antigen-antibody interaction by integrating the enzyme-labeled antibody technology, so that the purpose of visually analyzing the antigen-antibody interaction is achieved by using a method of visually analyzing the content of enzyme. The method is applied to the visual detection of relevant antigens and antibodies in clinical specimens. By virtue of the method, the detection of the antigen-antibody reaction is completed by only using naked eyes. The method and the application thereof have the advantages of high sensitivity, simplicity in operation, low cost and high 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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IPC IPC(8): G01N21/76G01N33/53G01N33/569
Inventor 蒋兴宇鲜于运雷王卓
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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