Biological probe and biological immunosensor for detecting bungarus multicinctus venom and method for detecting bungarus multicinctus venom based on proportional signal

An immune sensor and Bungare venom technology, which is applied in biological testing, biological material analysis, instruments, etc., can solve the problems of long detection time and high equipment cost, and achieve the effects of short detection time, high sensitivity and wide repeatability

Active Publication Date: 2022-02-15
YUNNAN UNIV
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  • Application Information

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Problems solved by technology

Chromatographic analysis has the advantage of fast analysis speed, but its detection takes a long time and the equipment cost is relatively high

Method used

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  • Biological probe and biological immunosensor for detecting bungarus multicinctus venom and method for detecting bungarus multicinctus venom based on proportional signal
  • Biological probe and biological immunosensor for detecting bungarus multicinctus venom and method for detecting bungarus multicinctus venom based on proportional signal
  • Biological probe and biological immunosensor for detecting bungarus multicinctus venom and method for detecting bungarus multicinctus venom based on proportional signal

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preparation example Construction

[0042] In the present invention, the preparation method of the detection bioprobe based on the ratio-type double signal of the bungalow snake venom comprises the following steps:

[0043] (1) Mix 1,3,5-tri(4-aminophenyl)benzene, 2,5-dimethoxybenzene-1,4-dicarbaldehyde, acetic acid with an organic solvent, and carry out solvothermal reaction to obtain a total Valence organic framework materials;

[0044] (2) the covalent organic framework material and alcohol solvent, HAuCl 4and reducing agent are mixed for reduction reaction to obtain AuNPs / COF nanocomposite material;

[0045] (3) Mixing the AuNPs / COF nanocomposite material with a sulfhydryl-modified bungalo β toxin aptamer to obtain a bioprobe for detecting bungale venom based on a ratiometric dual signal.

[0046] The present invention mixes 1,3,5-tris(4-aminophenyl)benzene, 2,5-dimethoxybenzene-1,4-dicarbaldehyde, acetic acid and an organic solvent for solvothermal reaction to obtain covalent Organic Frameworks (COFs). ...

Embodiment 1

[0105] A preparation method of a proportional chemical bioimmunosensor for detecting bununga venom, the specific steps are as follows:

[0106] (1) Preparation of AuNPs@COF: 0.030 mmol of 1,3,5-tris(4-aminophenyl)benzene and 0.030 mmol of 2,5-dimethoxybenzene-1,4-dicarbaldehyde were added together 4.5 mL of 1,4-dioxane-butanol-methanol (V 1 / V 2 / V 3 ,4:4:1) co-solvent system. Then, the mixture was sonicated for 20 min to obtain a homogeneous mixed solution. After adding 0.050 mL of 12M acetic acid aqueous solution dropwise, let stand at room temperature for 2 hours, then add 0.45 mL of 12M acetic acid aqueous solution, and heat at 70°C for 24 hours. Finally, the product was cooled to room temperature, and the synthesized COF was washed three times with THF and acetone, and dried in vacuum for 24 h. The obtained COF (0.060g) was added to 30mL of methanol, hydrochloric acid was added to adjust the pH to 4, and then 200 μL of 1% HAuCl was slowly added dropwise 4 4H 2 O in...

Embodiment 2

[0118] A preparation method of a proportional chemical bioimmunosensor for detecting bununga venom, the specific steps are as follows:

[0119] (1) Preparation of AuNPs@COF: 0.030 mmol of 1,3,5-tris(4-aminophenyl)benzene and 0.040 mmol of 2,5-dimethoxybenzene-1,4-dicarbaldehyde were added together 4.5 mL of 1,4-dioxane-butanol-methanol (V1 / V2 / V3, 4:4:1) co-solvent system. Then, the mixture was sonicated for 20 min to obtain a homogeneous mixed solution. After adding 0.050 mL of 12M acetic acid aqueous solution dropwise, let stand at room temperature for 2 hours, then add 0.45 mL of 12M acetic acid aqueous solution, and heat at 70°C for 24 hours. Finally, the product was cooled to room temperature, and the synthesized COF was washed three times with THF and acetone, and dried in vacuum for 24 h. Add the obtained COF (0.060g) into 30mL of methanol, add hydrochloric acid to adjust the pH to 4, then slowly add 200μL of 1% HAuCl4·4H2O into the mixed solution, stir at 0°C for 5h, ...

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Abstract

The invention provides a biological probe and a biological immunosensor for detecting bungarus multicinctus venom and a method for detecting coral snake venom based on proportional signals, and belongs to the technical field of biological immunosensing. AuNPs/COF serves as a signal amplification substance, the probe has a large number of catalytic active sites, and has a catalytic reduction effect and an enrichment effect on a guest signal molecule p-nitrophenol when the probe is used for detection by an ELISA (enzyme-linked immunosorbent assay), and a proportional chemical biological immunosensor constructed by the probe can realize accurate measurement of bungarus multicinctus venom, has the advantages of good stability, good repeatability, wide detection range, short detection time and low cost. According to the biological immunosensor, S-GQD serves as a reducing agent, the stability of the sensor can be effectively improved, meanwhile, the reducing capacity of the sensor can be catalyzed and improved by the gold nanoparticles, the S-GQD and the AuNPs/COF have the synergistic positive feedback amplification effect on signals, and therefore the sensitivity of the sensor is improved.

Description

technical field [0001] The invention relates to the technical field of bioimmune sensing, in particular to a detection bioprobe and a bioimmune sensor for detecting bungali venom and a method for detecting bungalo venom based on proportional signals. Background technique [0002] Corona snake is the fourth most poisonous terrestrial snake in the world, and it is also the most poisonous terrestrial snake in China. It is widely distributed in Anhui, Zhejiang, Jiangsu, Jiangxi, Fujian, Taiwan, Hubei, Hunan, Guangdong, Guangxi, and Hainan in China. , Guizhou, Yunnan and other provinces. Bungarotoxin is mainly composed of proteins and polypeptides, mainly neurotoxins, and has the characteristics of hidden symptoms, high toxicity, rapid onset, and low lethal dose. Therefore, timely diagnosis through specific and sensitive biomarkers and analytical tools can shorten the time required to find targeted antivenom, thereby effectively improving the survival rate. [0003] At present,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/58G01N33/553G01N33/543G01N33/532
CPCG01N33/68G01N33/588G01N33/54346G01N33/553G01N33/532G01N2333/4613
Inventor 李灿鹏王犁赵卉李文辉
Owner YUNNAN UNIV
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