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Protein chip for detecting seven markers of lung cancer and preparation method thereof

A protein chip and marker technology, applied in the biological field, can solve the problems of poor sensitivity and poor reproducibility, and achieve the effects of low price, lower detection cost, and improved detection efficiency.

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

[0020] Elisa is the most widely used in clinical applications, but this method can only detect one indicator at a time, and its sensitivity and reproducibility are not good

Method used

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  • Protein chip for detecting seven markers of lung cancer and preparation method thereof
  • Protein chip for detecting seven markers of lung cancer and preparation method thereof
  • Protein chip for detecting seven markers of lung cancer and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] A protein chip for detection of lung cancer markers, the preparation method of the protein chip comprises the following steps:

[0047] (1) Black slide pretreatment;

[0048] 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;

[0049] ② Soak black slides in 0.05% silane solution (25% ethanol) for 60 minutes;

[0050] ③Purge the soaked black glass slides with nitrogen gas, put them in an oven, and bake them at 180°C for 0.2h.

[0051] (2) Antigen solution spotting;

[0052] refer to figure 1 , using machine automation to spot GAGE7 antigen, MAGE A1 antigen, CAGE antigen, P53 antigen, PGP9.5 antigen, SOX2 antigen and GBU4-5 antigen solution, the concentration of the antigen solution is 0.02mg / mL, and each point is 20nL, each antigen point Sample distribution such as figure 1 shown.

[0053] (3) closed process;

[0054] The spotted black slide was submerged in blocking sol...

Embodiment 2

[0058] A protein chip for detection of lung cancer markers, the preparation method of the protein chip comprises the following steps:

[0059] (1) Black slide pretreatment;

[0060] 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;

[0061] ② Soak black slides in 0.5% silane solution (medium: 25% ethanol) for 30 minutes;

[0062] ③Purge the soaked black glass slides with nitrogen gas, put them in an oven, and bake them at 140°C for 0.5h.

[0063] (2) Antigen solution spotting;

[0064] refer to figure 1 , using machine automation to spot GAGE7 antigen, MAGE A1 antigen, CAGE antigen, P53 antigen, PGP9.5 antigen, SOX2 antigen and GBU4-5 antigen solution, the concentration of the antigen solution is 0.02mg / mL, 20nL for each spot, each antibody spot Sample distribution such as figure 1 shown. .

[0065] (3) closed process;

[0066] The spotted black slide was submerged in blocki...

Embodiment 3

[0070] A protein chip for detection of lung cancer markers, the preparation method of the protein chip comprises the following steps:

[0071] (1) Black slide pretreatment;

[0072] 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;

[0073] ② Soak black slides in 1% silane solution (medium: 25% ethanol) for 20 minutes;

[0074] ③Purge the soaked black glass slides with nitrogen gas, put them into an oven, and bake them at 100°C for 0.6h.

[0075] (2) Antigen solution spotting;

[0076] refer to figure 1 , using machine automation to spot GAGE7 antigen, MAGE A1 antigen, CAGE antigen, P53 antigen, PGP9.5 antigen, SOX2 antigen and GBU4-5 antigen solution, the concentration of the antigen solution is 0.02mg / mL, 20nL for each spot, each antibody spot Sample distribution such as figure 1 shown.

[0077] (3) closed process;

[0078] The spotted black slide was submerged in blocking ...

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Abstract

The invention discloses a protein chip for detecting seven items of lung cancer. 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 provided by the invention is used for detecting seven markers of lung cancer, and the unique protein chip technology of the company is applied to realize joint detection, so that the detection efficiency can be effectively improved, and the detection cost is reduced. The inventor develops a protein chip diagnostic kit simultaneously containing seven self-immune indexes including a GAGE7 antibody, an MAGE A1 antibody, a CAGE antibody, a P53 antibody, a PGP9.5 antibody, anSOX2 antibody and a GBU4-5 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 seven markers of lung cancer and a preparation method thereof. Background technique [0002] Currently, cancer is a major public health problem in most countries, including China. With the growth and aging of the world population, and the formation of risk factors such as smoking, obesity, and lack of physical activity, the incidence of cancer is also increasing year by year, seriously threatening human health. According to the latest national cancer report of the National Cancer Center in 2018, the estimated number of new cases of malignant tumors in the country in 2014 was 3.804 million. Ranked by the number of cases, lung cancer ranked first in the country. In developed countries, lung cancer is the leading cause of death among male patients and has surpassed breast cancer as the leading cause of death among female patients [1] . Although the penetr...

Claims

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Application Information

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IPC IPC(8): G01N33/531G01N33/552G01N33/574
CPCG01N33/531G01N33/552G01N33/57423G01N33/57484
Inventor 张倩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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