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Nanometer synergistic glycosyl group functionalized bacterial toxin molecular engram film 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 effects of good molecular recognition performance, strong anti-interference ability and strong specificity

Inactive Publication Date: 2010-03-17
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • Nanometer synergistic glycosyl group functionalized bacterial toxin molecular engram film substrate as well as preparation method and application thereof
  • Nanometer synergistic glycosyl group functionalized bacterial toxin molecular engram film substrate as well as preparation method and application thereof
  • Nanometer synergistic glycosyl group functionalized bacterial toxin molecular engram film substrate as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1 (diphtheria toxin)

[0041] A method for preparing a molecularly imprinted membrane substrate for detecting diphtheria toxin, comprising the following steps:

[0042] (1) Select the functional monomer A-D-pyranosylmannose that can synthesize MIPs with diphtheria toxin;

[0043] (2) Preparation of carbon nanotube solution: 2 mg of multi-walled carbon nanotubes (MWCNTs) were added to 1 ml of dimethyl sulfoxide solution under ultrasonic stirring to obtain a black suspension that was MWCNTs solution;

[0044] (3) Take 20 μl of the prepared MWCNTs solution and sonicate for 30 minutes to obtain a uniformly dispersed MWCNTs solution;

[0045] (4) Template molecule diphtheria toxin, functional monomer A-D-pyranosylmannose, cross-linking agent ethylene glycol dimethacrylate (EGDMA), porogen chloroform, initiator azobisisobutyronitrile, organic The solvent dichloromethane is mixed uniformly in a molar ratio of 0.1:2.5:0.5:70:0.05:2.0 to obtain a diphtheria toxin M...

Embodiment 2

[0052] Embodiment 2 (tetanus toxin)

[0053] A method for preparing a molecularly imprinted membrane substrate for detecting tetanus toxin, comprising the following steps:

[0054] (1) select the functional monomer sialyl ganglioside that can synthesize MIPs with tetanus toxin;

[0055] (2) Preparation of nano-gold solution: take 50mL 0.01% HAuCl 4 Heat to boiling, quickly add 1mL 1% sodium citrate solution for reduction, and the prepared bright rose red nano-gold solution is stored in a 4°C refrigerator for later use;

[0056] (3) Take 20 μl of the prepared nano-gold solution, and sonicate for 20 minutes to obtain a uniformly dispersed nano-gold solution;

[0057] (4) Template molecule tetanus toxin, functional monomer sialyl ganglioside, crosslinking agent trimethylolpropane trimethacrylate, porogen dichloromethane, initiator azobisisobutyronitrile, organic The solvent dichloromethane is mixed in a molar ratio of 0.5:2.5:4:50:0.1:2.5 to obtain a tetanus toxin MIPs solutio...

Embodiment 3

[0064] Embodiment 3 (botulinum toxin)

[0065] A method for preparing a molecularly imprinted membrane substrate for detecting botulinum toxin, comprising the following steps:

[0066] (1) Select the functional monomer N-acetylglucosamine that can synthesize MIPs with botulinum toxin;

[0067] (2) Preparation of nano-platinum solution: get 4ml of 5% H 2 PtCl 6 ·6H 2 The O solution was added to 340ml of double-distilled water, and then heated while stirring at 80°C. After adding 60ml of 1% sodium citrate solution, the resulting solution, namely the nano-platinum solution, was kept at 80±0.5°C for about 4 hours. The process is recorded by adsorption spectroscopy, when PtCl 6 2- When the adsorption band disappears, it indicates that the reaction is over, that is, the nano-platinum solution is obtained;

[0068] (3) Take 20 μl of the prepared nano-platinum solution and ultrasonicate for 40 minutes to obtain a uniformly dispersed nano-platinum solution;

[0069] (4) Template...

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Abstract

The invention relates to a nanometer synergistic glycosyl group functionalized bacterial toxin molecular engram film substrate as well as a preparation method and an application thereof. The nanometersynergistic bacterial toxin molecular engram film substrate is to form a plurality of layers of mutually-alternating nanometer materials and glycosyl group functionalized bacterial toxin molecular engram polymers on a gold quartz crystal substrate by taking the glycosyl group functionalized bacterial toxin molecular engram polymers as coupling agents. The preparation method is realized as follows: selecting sugar molecule functional monomers which can recognize the specificity of bacterial toxin and synthesize corresponding molecular engram polymers; preparing molecular engram polymer solution; and modifying the nanometer materials and the molecular engram polymers on the surface of the gold quartz crystal substrate. The molecular engram substrate is connected to a piezoelectric quartz crystal microbalance to detect the bacterial toxin in an environment sample extraction solution. The invention has simple operation and high specificity and sensitivity for detecting the bacterial toxin.

Description

technical field [0001] The invention relates to a nano-synergized sugar-based functionalized bacterial toxin molecularly 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 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N5/02
Inventor 黄加栋汪世华裴梅山于京华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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