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Treatment method for reducing non-specific adsorption in autoimmune antibody detection and kit thereof

An autoimmune antibody, non-specific technology, applied in the field of immune detection, can solve problems such as hindering non-specific adsorption, affecting detection results, non-specific adsorption, etc., to reduce non-specific adsorption capacity, reduce false positive probability, and improve detection rate. Effect

Pending Publication Date: 2020-08-11
四川携光生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The non-specific adsorption of magnetic particles to proteins in serum samples will reduce the detection sensitivity, and even cause false positives in the test results, affecting the test results
In actual detection, non-specific adsorption is unavoidable, and the sample contains a variety of proteins, which can easily cause non-specific adsorption
The main causes of non-specific adsorption are hydrophobic interaction and electrostatic interaction; at present, there are three main ways to reduce non-specific adsorption. 1. Use a blocking agent to block non-specific sites on the surface of magnetic particles, which hinders non-specific adsorption. Commonly used protein Blocking agents include BSA, casein, and bioblockers from different manufacturers; 2. Modify the surface of magnetic particles. At present, different manufacturers have improved the surface of magnetic particles to reduce the surface modification of non-specific adsorption. However, due to different serum samples , there are some differences in protein types, and it cannot fundamentally solve the problem

Method used

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  • Treatment method for reducing non-specific adsorption in autoimmune antibody detection and kit thereof
  • Treatment method for reducing non-specific adsorption in autoimmune antibody detection and kit thereof
  • Treatment method for reducing non-specific adsorption in autoimmune antibody detection and kit thereof

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Experimental program
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Effect test

Embodiment 1

[0028] During the specific implementation process of the detection kit for reducing autoimmune antibodies, the principle of an indirect method is used to measure the content of specific autoimmune antibody IgG in human serum and plasma samples.

[0029] This kit contains M, R1, R2, calibrator and supporting sample diluent, cleaning solution and substrate solution. One of the specific ratios of M, R1, R2, calibrator, and supporting sample diluent and cleaning solution is shown in Table 1.

[0030] Table 1 dsDNA reagent components

[0031]

[0032]

[0033] Note: In the dsDNA reagent component table above, add the magnetic bead shell polymer to the reagent R1 component.

[0034] Magnetic bead shell polymer particles can be added to any one of R1, M, R2, and sample diluent, with a concentration of 5ug-50ug / mL, which has the same effect.

[0035] The specific detection process of the kit is as follows:

[0036]Mix 20-50uL of the R1 reagent component with 20-50uL of the sa...

Embodiment 2

[0038] The proportioning mode of each reagent component in the kit provided above:

[0039] ① R1 reagent components: 0.1M Tris (pH 7.4, containing 1% BSA).

[0040] ②M reagent components: magnetic particles coupled with dsDNA antigen, including magnetic particles with avidin on the surface coupled with biotinylated antigen, and magnetic particles with carboxyl, amino, and tosyl groups on the surface directly coupled with antigen.

[0041] Specific method: Take biotinylated antigen-coupled avidin magnetic beads as an example

[0042] The ratio of biotinylated antigen and magnetic particle can be 1-20ug / 1mg. Wash the avidin magnetic bead mother solution with 0.01M PBS buffer containing 1% BSA for 3-5 times, set the volume to 5mg / mL, add biotinylated antigen to the magnetic particle solution, and react in a serum mixer After 30 minutes (25°C±3°C), wash three times with 0.01M PBS (containing 1% BSA), and dilute with 0.1M Tris until the working concentration of magnetic particles...

Embodiment 3

[0050] like figure 1 and figure 2 As shown, in the above kit, a shell polymer particle 1 is designed to compete with the surface of the magnetic particle. The shell polymer particles 1 and the magnetic particles have the same or similar substances. Magnetic particles include shell polymer particles 1, core layer Fe 2 o 3 2 and functional groups on the surface of magnetic particles 3.

[0051] The shell polymer particle 1 is to remove the magnetic particle core layer Fe 2 o 3 2 and 3 polymers of functional groups on the surface of magnetic particles.

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Abstract

The invention belongs to the technical field of immunodetection, and discloses a treatment method for reducing non-specific adsorption in autoimmune antibody detection and a kit thereof. The kit for reducing detection of the autoimmune antibody, provided by the invention, is formed by combining polymer particles, a magnetic particle coupling antigen, AP-labeled anti-human IgG, an autoantibody IgGantibody and a Tris buffer solution after mutual dissolution. According to the detection kit for reducing the autoimmune antibody, polymer particles and magnetic particles in the kit are added to competitively bind non-specific substances to a detection target object; the non-specific adsorption capacity of the magnetic beads on the surfaces of the magnetic particles is further reduced; the falsepositive probability of the detection kit caused by non-specific adsorption in the autoimmune antibody detection process is remarkably reduced, the influence of non-specific substances on a clinical sample test result is weakened, and the detection rate of the detection kit and the effectiveness and stability of the detection result are improved.

Description

technical field [0001] The invention belongs to the technical field of immune detection, and in particular relates to a treatment method for reducing non-specific adsorption in autoimmune antibody detection and a kit thereof. Background technique [0002] Chemiluminescence immunoassay (CLIA) technology is an immunoassay technology established in the 1970s. Detection and analysis techniques for antigens, antibodies, hormones, enzymes, fatty acids, vitamins and drugs. It is a newest immunoassay technique developed after radioimmunoassay, enzyme immunoassay, fluorescent immunoassay and time-resolved fluorescent immunoassay. It has the characteristics of high accuracy, no pollution, and fast detection speed. [0003] As a new type of carrier, nano-magnetic particles are widely used in the fields of diagnosis and food safety. Magnetic microspheres have the advantages of high magnetic content and large surface area. A large number of surface groups and modified proteins are atta...

Claims

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Application Information

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IPC IPC(8): G01N33/543
CPCG01N33/54326
Inventor 秦枫潘敬梅黄敬双张伟万文琴鲜静吴斌张小飞颜松何志威
Owner 四川携光生物技术有限公司
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