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Protein chip for detecting autoimmune disease markers and preparation method thereof

An autoimmune disease and protein chip technology, which is applied in the field of protein chips for detecting autoimmune disease markers and its preparation, can solve the problems of low detection efficiency, low cost, general sensitivity and specificity, etc. cost, and the effect of improving detection efficiency

Inactive Publication Date: 2018-11-16
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

Western blotting is suitable for primary screening of self-immunity. The method is simple, the sensitivity and specificity are average, it does not require complicated instruments, and the cost is low. Can realize qualitative observation; enzyme-linked immunosorbent immunoassay (ELISA) and single-index chemiluminescence are the most commonly used methods for immunodiagnosis, with high sensitivity and specificity, and the reliability of the results is higher than that of western blot. The project is tested separately, and the detection efficiency is low;

Method used

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  • Protein chip for detecting autoimmune disease markers and preparation method thereof
  • Protein chip for detecting autoimmune disease markers and preparation method thereof
  • Protein chip for detecting autoimmune disease markers and preparation method thereof

Examples

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Effect test

Embodiment 1

[0053] A protein chip for autoimmune disease marker detection, the preparation method of the protein chip comprises the following steps:

[0054] (1) Black slide pretreatment;

[0055] ① Soak the black slide in the slide pretreatment solution containing 2% NaOH for 16 hours, and then wash it with purified water for 2 to 8 times;

[0056] ② Soak black slides in 0.05% silane solution (25% ethanol) for 60 minutes;

[0057] ③Purge the soaked black glass slides with nitrogen gas, put them in an oven, and bake them at 180°C for 0.2h.

[0058] (2) Antibody solution spotting;

[0059] refer to figure 1 , use machine automation to spot SSA52 antigen, SSA60 antigen, SSB antigen, dsDNA antigen, U1-snRNP antigen, CENP-B antigen, Histone antigen, P0 antigen, Sm antigen, Nucleosome antigen, Scl-70 antigen and Jo-1 antigen solution , the concentration of the antigen solution is 0.1mg / mL, each spot is 20nL, and the spot distribution of each antibody is as follows: figure 1 shown. .

[...

Embodiment 2

[0065] A protein chip for autoimmune disease marker detection, the preparation method of the protein chip comprises the following steps:

[0066] (1) Black slide pretreatment;

[0067] ① Soak the black slide in the slide pretreatment solution containing 2% NaOH for 24 hours, and then wash it with purified water for 2 to 8 times;

[0068] ② Soak black slides in 0.5% silane solution (medium: 25% ethanol) for 30 minutes;

[0069] ③Purge the soaked black glass slides with nitrogen gas, put them in an oven, and bake them at 140°C for 0.5h.

[0070] (2) Antibody solution spotting;

[0071] refer to figure 1 , use machine automation to spot SSA52 antigen, SSA60 antigen, SSB antigen, dsDNA antigen, U1-snRNP antigen, CENP-B antigen, Histone antigen, P0 antigen, Sm antigen, Nucleosome antigen, Scl-70 antigen and Jo-1 antigen solution , the concentration of the antibody solution is 0.1mg / mL, each spot is 20nL, and the distribution of each antibody spot is as follows: figure 1 shown....

Embodiment 3

[0077] A protein chip for autoimmune disease marker detection, the preparation method of the protein chip comprises the following steps:

[0078] (1) Black slide pretreatment;

[0079] ① Soak the black slide in the slide pretreatment solution containing 2% NaOH for 20 hours, and then wash it with purified water for 2 to 8 times;

[0080] ② Soak black slides in 1% silane solution (medium: 25% ethanol) for 20 minutes;

[0081] ③Purge the soaked black glass slides with nitrogen gas, put them into an oven, and bake them at 100°C for 0.6h.

[0082](2) Antibody solution spotting;

[0083] refer to figure 1 , use machine automation to spot SSA52 antigen, SSA60 antigen, SSB antigen, dsDNA antigen, U1-snRNP antigen, CENP-B antigen, Histone antigen, P0 antigen, Sm antigen, Nucleosome antigen, Scl-70 antigen and Jo-1 antigen solution , the concentration of the antibody solution is 0.1mg / mL, each spot is 20nL, and the distribution of each antibody spot is as follows: figure 1 shown. ...

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Abstract

The invention discloses a protein chip for detecting autoimmune disease markers. A preparation method of the protein chip comprises the following steps: (1) pretreatment of black slide; (2) spotting of antigen solution; (3) blocking technology; preparation of the protein chip. The protein chip of the invention is used for detecting autoimmune disease markers. The joint detection achieved by the use of the company's original protein chip technology can effectively improve detection efficiency and reduce detection costs. The inventor has developed a protein chip diagnostic kit, which simultaneously includes anti-SSA52 antibody, anti-SSA60 antibody, anti-SSB antibody, anti-dsDNA antibody, anti-U1-snRNP antibody, anti-CENP-B antibody, anti-Histone antibody, anti-P0 antibody, anti-Sm antibody,anti-Nucleosome antibody, anti-Scl-70 antibody and anti-Jo-1 antibody. The kit has the advantages of speediness, 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 autoimmune disease markers and a preparation method thereof. Background technique [0002] Autoimmune diseases are diseases in which the immune system reacts 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 destroys or rejects these foreign substances. Under the influence of certain factors, some abnormalities appear 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 against some of the body's own components and active lymphocytes, which will damage and destroy its own tissues and organs, leading to disease. If left unchecked, the conseq...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 施启尧丁俊杰孙裔雷泮锋纲
Owner 江苏三联生物工程股份有限公司
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