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Protein chip for detecting heart failure marker and preparation method thereof

A technology of protein chips and markers, applied in the biological field, can solve the problems of slow detection speed, high detection cost, waste of patient burden, etc., and achieve the effect of reducing detection cost, improving detection efficiency and low cost

Inactive Publication Date: 2018-08-17
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

Although these two markers can be determined, there are disadvantages such as slow detection speed and high detection cost
Moreover, there are practical problems such as inconvenient operation and high error probability in the separate detection of the two indicators.
Heart failure patients are generally emergency patients. In order to speed up the detection, many hospitals have to collect multiple blood samples at the same time for the detection of ST2 and NT-proBNP respectively, resulting in a large waste and patient burden

Method used

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  • Protein chip for detecting heart failure marker and preparation method thereof
  • Protein chip for detecting heart failure marker and preparation method thereof
  • Protein chip for detecting heart failure marker and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] A protein chip for detection of heart failure markers, the preparation method of the protein chip comprises the steps of:

[0031] (1) Black slide pretreatment;

[0032] 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;

[0033] ② Soak the black slide in 0.05% silane aqueous solution for 60 minutes;

[0034] ③Put the soaked black glass slide into an oven and bake at 180°C for 0.2h.

[0035] (2) Antibody solution spotting;

[0036] refer to figure 1 , the ST2 monoclonal antibody solution and NT-proBNP monoclonal antibody solution were automatically spotted by machine, the concentration of the antibody solution was 0.1mg / mL, and 20nL was spotted for each spot. The spot distribution of each antibody was as follows: figure 1 shown. .

[0037] (3) closed process;

[0038] The spotted black slide was submerged in blocking solution (PBS buffer containing 1% bovine serum album...

Embodiment 2

[0042] A protein chip for detection of heart failure markers, the preparation method of the protein chip comprises the following steps:

[0043] (1) Black slide pretreatment;

[0044] 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;

[0045] ② Soak the black slide in 0.5% silane aqueous solution for 30 minutes;

[0046] ③Put the soaked black glass slide into an oven and bake at 140°C for 0.5h.

[0047] (2) Antibody solution spotting;

[0048] refer to figure 1 , the ST2 monoclonal antibody solution and NT-proBNP monoclonal antibody solution were automatically spotted by machine, the concentration of the antibody solution was 0.1mg / mL, and 20nL was spotted for each spot. The spot distribution of each antibody was as follows: figure 1 shown. .

[0049] (3) closed process;

[0050] The spotted black slide was submerged in blocking solution (PBS buffer containing 2% ovalbumin) f...

Embodiment 3

[0054]A protein chip for detection of heart failure markers, the preparation method of the protein chip comprises the steps of:

[0055] (1) Black slide pretreatment;

[0056] 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;

[0057] ② Soak the black slide in 1% silane aqueous solution for 20 minutes;

[0058] ③Put the soaked black glass slide into an oven and bake at 100°C for 0.6h.

[0059] (2) Antibody solution spotting;

[0060] refer to figure 1 , the ST2 monoclonal antibody solution and NT-proBNP monoclonal antibody solution were automatically spotted by machine, the concentration of the antibody solution was 0.1mg / mL, and 20nL was spotted for each spot. The spot distribution of each antibody was as follows: figure 1 shown. .

[0061] (3) closed process;

[0062] The spotted black slide was submerged in blocking solution (Tris buffer containing 3% bovine serum albumin)...

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Abstract

The invention discloses a protein chip for detecting a heart failure marker, and a preparation method of the protein chip comprises the following steps: (1) black slide pretreatment; (2) antibody solution sample application; and (3) sealing process to obtain the protein chip. The protein chip is used for detecting the heart failure marker, by combination with the company's original protein chip technology, the protein chip can effectively improve the detection efficiency and reduce the detection cost; a protein chip diagnostic kit including two indexes of ST2 and NT-proBNP is also disclosed, and the protein chip diagnostic kit has the advantages of fast, high efficiency and low cost.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a protein chip for detecting heart failure markers and a preparation method thereof. Background technique [0002] The mortality rate of cardiovascular disease is higher than other diseases including cancer and AIDS, and it is the "first killer" of human health. Traditional diagnostic techniques, such as electrocardiography, cardiovascular color Doppler ultrasound, nuclear magnetic resonance, and heart failure enzyme spectrum, play an important role in the diagnosis of cardiovascular diseases, and have been adopted and continuously improved. With the continuous development of science and technology, the detection of heart failure markers in blood has been gradually applied clinically, providing a method for early diagnosis and prognosis of cardiovascular diseases. [0003] Two important heart failure markers: ST2 and NT-proBNP. ST2 (scientific name soluble growth-stimulating expres...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6803G01N2800/325
Inventor 泮锋纲施启尧丁俊杰成舜
Owner 江苏三联生物工程股份有限公司
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