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Protein chip for myasthenia gravis marker detection and preparation method thereof

A myasthenia gravis and protein chip technology, applied in the biological field, can solve the problems of short product shelf life, inconvenient operation, cumbersome use, etc., and achieve the effect of realizing automatic detection, improving detection efficiency and reducing detection cost

Pending Publication Date: 2020-04-10
江苏三联生物工程股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the radioactivity and inconvenient operation of RIA, the short shelf life of products has been basically eliminated; the sensitivity and specificity of enzyme-linked immunosorbent immunoassay (ELISA) are relatively poor, and it can only be semi-quantitative.
Moreover, ELISA is a single-index one-by-one measurement, which is cumbersome to use and difficult to automate.
Products that have not been commercialized so far
At present, there is no report on the application of protein chip technology to the detection of autoantibodies in myasthenia gravis

Method used

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  • Protein chip for myasthenia gravis marker detection and preparation method thereof
  • Protein chip for myasthenia gravis marker detection and preparation method thereof
  • Protein chip for myasthenia gravis marker detection and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] A protein chip for the detection of myasthenia gravis autoantibodies, the preparation method of the protein chip comprises the following steps:

[0050] (1) Pretreatment of black glass slides;

[0051] ① Soak the black glass slides in the slide pretreatment solution containing 2% NaOH for 16 hours, and then wash them with purified water for 2 to 8 times;

[0052] ② Soak the black glass slide in 0.05% silane solution (25% ethanol) for 60min;

[0053] ③Purge the soaked black glass slides with nitrogen and put them in an oven, and bake them at 180℃ for 0.2h.

[0054] (2) Spotting the antigen solution;

[0055] refer to figure 1 , using machine automated spotting AchR antigen, MusK antigen, Titin antigen, LRP4 antigen, RyR antigen, AchE antigen, Kv1.4 antigen and GQ1b antigen, the concentration of antigen solution is 0.01mg / mL, each spotting 20nL, each antigen spotting distributed as figure 1 shown.

[0056] (3) closed process;

[0057] The spotted black glass slide ...

Embodiment 2

[0061] A protein chip for the detection of myasthenia gravis autoantibodies, the preparation method of the protein chip comprises the following steps:

[0062] (1) Pretreatment of black glass slides;

[0063] ① Soak the black glass slides in the slide pretreatment solution containing 2% NaOH for 24 hours, and then wash them with purified water for 2 to 8 times;

[0064] ② Soak the black glass slides in 0.5% silane solution (the medium is 25% ethanol) for 30min;

[0065] ③Purge the soaked black glass slides with nitrogen and put them in an oven, and bake them at 140°C for 0.5h.

[0066] (2) Spotting the antigen solution;

[0067] refer to figure 1 , using machine automated spotting of AchR antigen, MusK antigen, Titin antigen, LRP4 antigen, RyR antigen, AchE antigen, Kv1.4 antigen and GQ1b antigen solution, the concentration of antigen solution is 0.01mg / mL, each spot spotting 20nL, each antigen spot distribution like figure 1 shown.

[0068] (3) closed process;

[0069]...

Embodiment 3

[0073] A protein chip for the detection of myasthenia gravis autoantibodies, the preparation method of the protein chip comprises the following steps:

[0074] (1) Pretreatment of black glass slides;

[0075] ① Soak the black glass slides in the slide pretreatment solution containing 2% NaOH for 20 hours, and then wash with purified water for 2 to 8 times;

[0076] ② Soak the black glass slide in 1% silane solution (the medium is 25% ethanol) for 20min;

[0077] ③Purge the soaked black glass slides with nitrogen and put them in an oven, and bake them at 100°C for 0.6h.

[0078] (2) Spotting the antigen solution;

[0079] refer to figure 1 , using machine automated spotting of AchR antigen, MusK antigen, Titin antigen, LRP4 antigen, RyR antigen, AchE antigen, Kv1.4 antigen and GQ1b antigen solution, the concentration of antigen solution is 0.01mg / mL, each spot spotting 20nL, each antigen spot distribution like figure 1 shown.

[0080] (3) closed process;

[0081] The spo...

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PUM

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Abstract

The invention discloses a protein chip for detecting myasthenia gravis markers. A preparation method of the protein chip comprises the following steps: (1) pretreating a black slide; (2) performing sample application of an antigen solution; (3) performing a sealing process; and preparing the protein chip. The protein chip is used for detecting myasthenia gravis markers, and the protein chip technology originally created by the company is applied to achieve combined detection, so that the detection efficiency can be effectively improved, and the detection cost is reduced. The inventor developsa protein chip diagnostic kit simultaneously containing eight indexes, namely an anti-AchR antibody, an anti-MusK antibody, an anti-Titin antibody, an anti-LRP4 antibody, an anti-RyR antibody, an anti-AchE antibody, an anti-Kv1.4 antibody and an anti-GQ1b antibody, and the protein chip diagnostic kit has the advantages of rapidness, high efficiency, low cost and the like.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a protein chip for detecting markers of myasthenia gravis and a preparation method thereof. Background technique [0002] Autoimmune diseases are diseases caused by the immune system's immune response to the components of its own body, causing damage. Under normal circumstances, the immune system only responds to foreign substances that invade the body, such as bacteria, viruses, parasites, and transplants, and eliminates or rejects these foreign substances. Under the influence of certain factors, some abnormalities have occurred in the tissue components of the body or the immune system itself, causing the immune system to mistake its own components as foreign objects to attack. At this time, the immune system will produce antibodies and active lymphocytes against some of the body's own components, which will damage and destroy its own tissues and organs, resulting in disease. If n...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/531G01N33/552G01N33/564
CPCG01N33/531G01N33/552G01N33/564G01N33/6893G01N2800/24
Inventor 王慧泮锋纲丁俊杰施启尧
Owner 江苏三联生物工程股份有限公司
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