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Glycosyl functional bacterial toxin molecularly imprinted membrane substrate as well as preparation method and application thereof

A molecularly imprinted membrane and bacterial toxin technology, applied in the field of analytical chemistry, can solve problems such as difficulty in direct application, and achieve the effect of high-throughput screening, high sensitivity, and improved sensitivity and accuracy

Inactive Publication Date: 2010-03-10
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this preparation method is difficult to directly apply to the gold substrate surface of most sensors.

Method used

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  • Glycosyl functional bacterial toxin molecularly imprinted membrane substrate as well as preparation method and application thereof
  • Glycosyl functional bacterial toxin molecularly imprinted membrane substrate as well as preparation method and application thereof
  • Glycosyl functional bacterial toxin molecularly imprinted membrane substrate as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1 (diphtheria toxin)

[0035] A method for preparing a glycosyl-functionalized diphtheria toxin molecularly imprinted membrane substrate, comprising the following steps:

[0036] (1) Select the functional monomer A-D-pyranosylmannose that can synthesize molecularly imprinted polymers with diphtheria toxin;

[0037] (2) template molecule diphtheria toxin, functional monomer A-D-pyranosylmannose, cross-linking agent ethylene glycol dimethacrylate (EGDMA), porogen chloroform, initiator azobisisobutyronitrile, The organic solvent dichloromethane is mixed uniformly at a molar ratio of 0.1:2.5:0.5:70:0.05:2.0 to obtain a glycosyl-functionalized diphtheria toxin molecularly imprinted polymer solution;

[0038] (3) Clean the gold quartz crystal substrate of the piezoelectric quartz crystal microbalance with deionized water, then soak it with pure methanol for 1 hour, then rinse it with deionized water for 3 times, then clean it with ultrasonic waves for 3 minutes, and...

Embodiment 2

[0042] Embodiment 2 (tetanus toxin)

[0043] A method for preparing a glycosyl-functionalized tetanus toxin molecularly imprinted membrane substrate, comprising the following steps:

[0044] (1) Select the functional monomer sialyl ganglioside that can synthesize molecularly imprinted polymers with tetanus toxin;

[0045] (2) Template molecule tetanus toxin, functional monomer sialyl ganglioside, crosslinking agent trimethylolpropane trimethacrylate, porogen dichloromethane, initiator azobisisobutyronitrile, The organic solvent dichloromethane is mixed uniformly at a molar ratio of 0.5:2.5:4:50:0.1:2.5 to obtain a glycosyl-functionalized tetanus toxin molecularly imprinted polymer solution;

[0046] (3) Clean the gold quartz crystal substrate of the piezoelectric quartz crystal microbalance with deionized water, then soak it with pure methanol for 1 hour, then rinse it with deionized water for 3 times, then clean it with ultrasonic waves for 3 minutes, and dry it to ensure T...

Embodiment 3

[0050] Embodiment 3 (botulinum toxin)

[0051] A method for preparing a glycosyl-functionalized botulinum toxin molecularly imprinted membrane substrate, comprising the following steps:

[0052] (1) Select N-acetylglucosamine, a functional monomer capable of synthesizing molecularly imprinted polymers with botulinum toxin;

[0053] (2) template molecule botulinum toxin, functional monomer N-acetylglucosamine, cross-linking agent N, N-methylenebisacrylamide, porogen methanol, initiator azobisisobutyronitrile, organic solvent Dichloromethane is mixed uniformly at a molar ratio of 2:2.5:5:75:0.1:15 to obtain a glycosyl-functionalized botulinum toxin molecularly imprinted polymer solution;

[0054] (3) Clean the gold quartz crystal substrate of the piezoelectric quartz crystal microbalance with deionized water, then soak it with pure methanol for 1 hour, then rinse it with deionized water for 3 times, then clean it with ultrasonic waves for 3 minutes, and dry it to ensure The su...

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Abstract

The invention relates to a glycosyl functional bacterial toxin molecularly imprinted membrane substrate as well as a preparation method and an application thereof. The molecularly imprinted membrane substrate forms a reaction layer on the surface of a gold quartz crystal substrate by taking a glycosyl functional bacterial toxin molecularly imprinted polymer as a recognition element. The preparation method comprises the following steps: selecting a carbohydrate molecule functional monomer which can carry out specificity recognition with bacterial toxin and compound the corresponding molecularly imprinted polymer; preparing a glycosyl functional molecularly imprinted polymer solution; and modifying the glycosyl functional molecularly imprinted polymer on the surface of the gold quartz crystal substrate by a substrate surface modifying technology to form a reaction layer. The glycosyl functional molecularly imprinted membrane substrate prepared by the method is connected to a piezoelectric quartz crystal microbalance to detect bacterial toxin in an environmental sample extracting solution. The molecularly imprinted membrane substrate has favorable molecular recognition performance and greatly improves the sensitivity and the selectivity for detecting the bacterial toxin.

Description

technical field [0001] The invention relates to a glycosyl functionalized bacterial toxin molecule imprinted membrane substrate and a preparation method and application thereof, belonging to the technical field of analytical chemistry. Background technique [0002] Bacterial toxins synthesized by many bacterial pathogens are primary agents of disease. This ability to produce bacterial toxins is an underlying mechanism of disease in many bacterial pathogens. There are three main types of bacterial toxins: exotoxins, endotoxins, and nonprotein toxins. Exotoxin is a typical water-soluble protein. It is known to be the most toxic bacterial toxin that can infect humans and is secreted by bacteria during the exponential growth phase. It is highly toxic at very low concentrations. Among them, the pathogenic mechanism of cholera toxin is to find the G protein as a cell recognition object, and then bind an ADP molecule to it. This switches the G protein into a continuously active ...

Claims

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Application Information

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IPC IPC(8): G01N27/327
Inventor 黄加栋汪世华于京华裴梅山葛慎光张秀明邢宪荣贺晓蕊林青朱晗孙纳新宋晓妍袁靓
Owner UNIV OF JINAN
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