Detection method of cytokine TNF-α based on aptamer and gold magnetic nanoparticles
A technology of gold magnetic nanoparticles and cytokines, applied in measuring devices, material analysis through electromagnetic means, instruments, etc., can solve the problems of high price and complicated operation, achieve low cost, high sensitivity, and realize the effect of recycling
Active Publication Date: 2019-06-25
SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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Biological detection methods involve the selection and cultivation of sensitive and specific target cells, and the operation is complicated; immunological detection methods require the use of antibodies, which are relatively expensive
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[0033] The present invention will be further described in detail below in conjunction with the accompanying drawings, so that those skilled in the art can implement it with reference to the description.
[0034] This case lists the detection method of the cytokine TNF-α of an embodiment, specifically may include:
[0035] 1) Use a magnetic glassy carbon electrode (diameter 2mm) as the working electrode. The electrode can preferably be placed in a newly prepared piranha solution (98% H 2 SO 4 : 30%H 2 o 2 =3:1) for 5 minutes to remove the substances adsorbed on the electrode surface.
[0036] 2) After washing with distilled water, polish the electrode with silicon carbide sandpaper (3000 mesh) until the mirror surface is smooth. Then, the electrodes were ultrasonically cleaned in ethanol and distilled water for 5 minutes each. Next, dry in a nitrogen atmosphere before use.
[0037] 3) Treated electrode with 0.5M H 2 SO 4 solution for electrochemical cleaning. Then, th...
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Abstract
The invention relates to a cytokine TNF-alpha detection method based on an aptamer and gold magnetic nanoparticles. The method comprises the following steps: using a magnetic glassy carbon electrode as a working electrode; modifying Fe3O4 nanoparticles with gold to prepare the gold magnetic nanoparticles; modifying the surfaces of the gold magnetic nanoparticles with a DNA probe 1; carrying out a hybridization reaction on a DNA probe 2 and the DNA probe 1 on the surfaces of the gold magnetic nanoparticles; adsorbing the gold magnetic nanoparticles to the surface of the glassy carbon electrode through magnetic field; and inserting the magnetic glassy carbon electrode to a liquid to be detected, acquiring a corresponding electrochemical response value, and calculating the concentration of the cytokine TNF-alpha in the liquid to be detected according to the corresponding relation between the electrochemical response value and the concentration of the cytokine TNF-alpha. The method has the characteristics of high sensitivity, fastness, low cost, realization of the application of TNF-alpha bio-sensing, and realization of fixation of the aptamer and cycle use of the electrode due to the introduction of a gold magnetic composite nanomaterial.
Description
technical field [0001] The invention relates to a detection method of cytokine TNF-α, in particular to a detection method of cytokine TNF-α based on aptamer and gold magnetic nanoparticles. Background technique [0002] Tumor necrosis factor (tumor necrosis factor, TNF) is a protein that appears in serum and can cause hemorrhagic necrosis of various tumors. TNF is mainly produced by activated macrophages, NK cells and T lymphocytes. In 1985, Shalaby named the TNF secreted by macrophages as TNF-α. TNF-α is a small molecular protein that can exert various biological effects after binding to specific receptors, including immune activation, inflammatory response, activation of cell growth-related gene expression, killing or inhibiting tumor cells, etc. Studies have shown that TNF-α can significantly increase liver cell damage and lead to kidney damage, reduce the prognosis of patients with liver cirrhosis and spontaneous peritonitis, and significantly increase mortality. TNF-...
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IPC IPC(8): G01N27/327
CPCG01N27/3277G01N27/3278
Inventor 缪鹏唐玉国陈锡峰杨大威郭振振
Owner SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI