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Method for performing immunological test on biomolecules by avidin/streptavidin magnetic composite particles

A technology for immunological detection and composite microparticles, which is applied in the field of immunoassay and can solve the problems of less labeling, volatile inactivation of biomolecules, etc.

Active Publication Date: 2013-04-10
XIAN GOLDMAG NANOBIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method solves the technical problems such as the small amount of labeling and the variability and inactivation of biomolecules in previous detection technologies.

Method used

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  • Method for performing immunological test on biomolecules by avidin/streptavidin magnetic composite particles
  • Method for performing immunological test on biomolecules by avidin/streptavidin magnetic composite particles
  • Method for performing immunological test on biomolecules by avidin/streptavidin magnetic composite particles

Examples

Experimental program
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Effect test

Embodiment 1

[0051] (1) Coupling of biotinylated anti-HBsAg antibody to avidin / streptavidin magnetic composite particles

[0052] Dissolve 200 μl biotinylated anti-HBsAg antibody in 400 μl pH=7.4 Tris-HCl coupling buffer, mix well and add to avidin / streptavidin magnetic composite particles, at 37°C, 180rpm, in a shaker React for 20 minutes, remove the magnetic separation after the reaction is complete, discard the supernatant; add 400 μl 1×PBS, magnetically separate, discard the supernatant;

[0053] (2) closed

[0054] Add 1ml of Tris-HCl buffer containing 5% skimmed milk powder and free biotin to the avidin / streptavidin magnetic composite particles, react at 37°C, 180rpm, in a shaker for 1 hour, magnetically separate, and discard the supernatant . Wash 3 times with 2ml pH=7.4, 1×PBS washing buffer, suspend the magnetic particles in 1ml pH=7.4, 0.02M Tris-HCl storage buffer, and store at 4°C for later use.

[0055] See figure 1 shown.

[0056] (3) Combined with the analyte

[0057] ...

Embodiment 2

[0074] (1) Coupling of biotinylated HCV antigen to avidin / streptavidin magnetic composite particles

[0075] Dissolve biotinylated HCV antigen in 400 μl pH=7.4 acetate coupling buffer, mix well, add to avidin / streptavidin magnetic composite particles, react at 37°C, 180rpm, in a shaker for 20min, After the reaction, remove the magnetic separation, discard the supernatant; add 400 μl 1×PBS, magnetic separation, discard the supernatant;

[0076] (2) closed

[0077] Add 1ml of Tris-HCl buffer containing 5% skimmed milk powder and free biotin to the avidin / streptavidin magnetic composite particles, react at 37°C, 180rpm, in a shaker for 1 hour, magnetically separate, and discard the supernatant . Wash 3 times with 2ml pH=7.4, 1×PBS washing buffer, suspend the magnetic particles in 1ml pH=7.4, 1×PBS storage buffer, and store at 4°C for later use.

[0078] (3) Combined with the analyte

[0079] The above-prepared avidin / streptavidin magnetic composite particles coupled with biot...

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PUM

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Abstract

The invention relates to the immune analysis field, in particular relating to a method for performing an immunological test on biomolecules by avidin / streptavidin magnetic composite particles, which takes avidin / streptavidin magnetic composite particles as carriers, and comprises the following steps:, coating biotinylated antibody or antigen, adding a specimen to be tested and enzyme labeled antibody / enzyme labeled antigen / enzyme labeled anti-antibody, adding chromogenic substrate and testing.Biotin and avidin / streptavidin has strong binding capacity, can fix the antigen / antibody labeled by the biotin through the avidin / streptavidin labeled on the surface of the magnetic composite particles so as to use the system to test specific antibody / antigen. The test system has the advantages of high sensitivity, high specificity, high stability, strong adaptability and the like.

Description

technical field [0001] The invention relates to the field of immune analysis, in particular to a method for immunological detection of biomolecules by avidin / streptavidin magnetic composite particles. Background technique [0002] The biotin-avidin system (BAS) is a biological reaction amplification system developed in the late 1970s. The principle is based on the following characteristics between biotin and avidin: (1) The two have a high specific affinity. Biotin is a small molecular substance; avidin includes avidin (also known as avidin) and streptavidin. The affinity between biotin and avidin / streptavidin is at least 10,000 times that of antigen (Ag) antibody (Ab), and it is not easily disturbed by the outside world, and the complex is stable. (2) Bridging effect. Both can couple biologically active substances such as proteins, nuclei, polysaccharides and enzymes, and can also be combined with solid phase materials. These properties can be used to couple them. (3) M...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543G01N33/576
Inventor 崔亚丽杨璐陈超耿婷婷
Owner XIAN GOLDMAG NANOBIOTECH
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