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Alpha-bungarus multicinctus toxin detection probe and method for non-diagnostic purpose detection of alpha-bungarus multicinctus toxin

A technology of bungalotoxin and detection probe, which is applied in the field of biosensing and can solve the problems of easy aggregation and limited application of nanozymes

Pending Publication Date: 2022-04-15
YUNNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, existing nanozymes are prone to agglomeration in water, which limits their application in biosensors.

Method used

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  • Alpha-bungarus multicinctus toxin detection probe and method for non-diagnostic purpose detection of alpha-bungarus multicinctus toxin
  • Alpha-bungarus multicinctus toxin detection probe and method for non-diagnostic purpose detection of alpha-bungarus multicinctus toxin
  • Alpha-bungarus multicinctus toxin detection probe and method for non-diagnostic purpose detection of alpha-bungarus multicinctus toxin

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preparation example Construction

[0041] The present invention has no special requirements on the source of the hydrogen-bonded organic framework material, and uses a hydrogen-bonded organic framework material whose organic ligand is 1,3,6,8-tetrakis(4-carboxyphenyl)pyrene, which is commercially available conventionally in the field. Or make your own. When preparing the hydrogen-bonded organic framework material by itself, the preparation method preferably includes:

[0042] In an alkaline environment, mix 4-methoxycarbonylphenylboronic acid, 1,3,6,8-tetrabromopyrene and a catalyst for a coupling reaction to obtain 1,3,6,8-tetrakis(4-(methyl Oxycarbonyl)phenyl)pyrene;

[0043] In an alkaline environment, the 1,3,6,8-tetrakis(4-(methoxycarbonyl)phenyl)pyrene is hydrolyzed and acidified to obtain 1,3,6,8-tetrakis(4-carboxybenzene) Pyrene;

[0044] The 1,3,6,8-tetrakis(4-carboxyphenyl)pyrene undergoes a self-assembly reaction to obtain a hydrogen-bonded organic framework material (HOF).

[0045] The present inv...

Embodiment 1

[0110] (1) Preparation of α-bungarotoxin detection probe

[0111] (1) First, 5g of 4-methoxycarbonylphenylboronic acid, 2.85g of 1,3,6,8-tetrabromopyrene, 0.1g of tetrakis(triphenylphosphine)palladium and 6g of potassium carbonate were dissolved in 100mL of dioxane, in N 2 Stir at 85°C for 72h under protection. Then the reaction product was poured into a solution of ice water and concentrated hydrochloric acid (v / v=3:1), the organic phase was collected by extraction with chloroform, finally dried with magnesium sulfate and dried in vacuo to remove the organic solvent, and finally obtained the product 1,3 ,6,8-Tetrakis(4-(methoxycarbonyl)phenyl)pyrene;

[0112] (2) First, dissolve 1,3,6,8-tetrakis(4-(methoxycarbonyl)phenyl)pyrene in a mixed solution of 100mL tetrahydrofuran / dioxane / water (v / v=5:2:2) 1 g KOH was then added, and the mixture was refluxed and stirred at 85° C. for 12 h, and the organic solvent was removed by vacuum drying. Then add 100mL H 2 O was stirred at r...

Embodiment 2

[0130] (1) Preparation of α-bungarotoxin detection probe

[0131] (1) First, 5g of 4-methoxycarbonylphenylboronic acid, 2.85g of 1,3,6,8-tetrabromopyrene, 0.1g of tetrakis(triphenylphosphine)palladium and 6g of potassium carbonate were dissolved in 100mL of dioxane, in N 2 Stir at 85°C for 72h under protection. Then the reaction product was poured into a solution of ice water and concentrated hydrochloric acid (v / v=3:1), the organic phase was collected by extraction with chloroform, finally dried with magnesium sulfate and dried in vacuo to remove the organic solvent, and finally obtained the product 1,3 ,6,8-Tetrakis(4-(methoxycarbonyl)phenyl)pyrene;

[0132] (2) First, dissolve 1,3,6,8-tetrakis(4-(methoxycarbonyl)phenyl)pyrene in a mixed solution of 100mL tetrahydrofuran / dioxane / water (v / v=5:2:2) 1 g KOH was then added, and the mixture was refluxed and stirred at 85° C. for 12 h, and the organic solvent was removed by vacuum drying. Then add 100mL H 2 O was stirred at r...

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Abstract

The invention provides an alpha-bungarus multicinctus toxin detection probe and a method for detecting alpha-bungarus multicinctus toxin with a non-diagnostic purpose, and belongs to the technical field of biosensing. A hydrogen bond organic framework material composed of 1, 3, 6, 8-tetra (4-carboxybenzene) pyrene is used as an enzyme material for catalyzing color development of a color developing agent, the surface of the hydrogen bond organic framework material has abundant carboxyl groups, the hydrogen bond organic framework material can be chemically combined with an aptamer of alpha-bungarus multicinctus toxin modified with amino through a CO-NH bond, and the stability of the probe is improved. The invention provides a method for detecting alpha-bungarus multicinctus toxin with a non-diagnostic purpose, an alpha-bungarus multicinctus toxin detection probe is used for specifically recognizing alpha-bungarus multicinctus toxin and catalyzing a chromogenic substrate to develop color, along with the increase of the concentration of the alpha-bungarus multicinctus toxin, the absorbance detected by ultraviolet analysis is enhanced, and the concentration of the alpha-bungarus multicinctus toxin is determined to be higher than the concentration of the alpha-bungarus multicinctus toxin. And the alpha-bungarus multicinctus toxin has a good linear relationship in the concentration range of the alpha-bungarus multicinctus toxin of 0.0001 to 316 ng mL <-1 >.

Description

technical field [0001] The invention relates to the technical field of biosensing, in particular to an alpha-bungarotoxin detection probe and a method for non-diagnostic detection of alpha-bungarotoxin. Background technique [0002] Coral snake is the fourth most venomous snake on land and the most poisonous snake in my country. 1 mg of venom in an adult coral snake can kill more than a dozen people. The toxins contained in the venom of the silver ring snake are mainly composed of proteins and polypeptides, mainly neurotoxins, including α-bungarotoxin (α-BGT), β-bungarotoxin (β-BGT), κ-silver Bungarotoxin (κ-BGT), γ-bungarotoxin (γ-BGT) and enzymes such as phospholipase A, etc. When bitten by a silver ring snake, you will not feel pain but drowsiness. When you are slightly poisoned, you will experience local paralysis. If the toxin acts on the neuromuscular junction, it will block the nerve conduction route, causing the striated muscles to fail to contract normally, leading...

Claims

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Application Information

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IPC IPC(8): G01N21/31G01N21/78
CPCY02A50/30
Inventor 李灿鹏曾靖赵卉李文辉
Owner YUNNAN UNIV
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