Preparation method and application of nano-synergistic carbohydrate functionalized molecularly imprinted column for detecting bacterial toxins

A molecularly imprinted column and bacterial toxin technology, applied in the direction of chemical reaction of materials for analysis, chemiluminescence/bioluminescence, etc., can solve the problems of common cross-reactions, achieve high sensitivity and detection range, high sensitivity, and operation quick and easy effects

Inactive Publication Date: 2010-04-07
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the main difficulties that limit the detection and research of bacterial toxins are: there are many structural similarities, and cross-reactivity is common; toxin detection is more easily affected by other interfering substances that are difficult to remove in the extract, etc.

Method used

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  • Preparation method and application of nano-synergistic carbohydrate functionalized molecularly imprinted column for detecting bacterial toxins
  • Preparation method and application of nano-synergistic carbohydrate functionalized molecularly imprinted column for detecting bacterial toxins
  • Preparation method and application of nano-synergistic carbohydrate functionalized molecularly imprinted column for detecting bacterial toxins

Examples

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Embodiment 1

[0036] A method for preparing a diphtheria toxin glycosyl functionalized molecularly imprinted column, comprising the following steps:

[0037] (1) Select functional monomers capable of synthesizing glycosyl-functionalized molecularly imprinted polymers with diphtheria toxin;

[0038] (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;

[0039] (3) Take 20 μl of the prepared MWCNTs solution and sonicate for 30 minutes to obtain a uniformly dispersed MWCNTs solution;

[0040] (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 at a molar ratio of 0.1:2.5:0.5:70:0.05:2.0 to obtain a diphtheria toxin glycosyl functi...

Embodiment 2

[0048] A method for preparing a tetanus toxin glycosyl functionalized molecularly imprinted column, comprising the following steps:

[0049] (1) Select the functional monomer sialyl ganglioside that can synthesize molecularly imprinted polymers with tetanus toxin;

[0050] (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;

[0051] (3) Take 20 μl of the prepared nano-gold solution, and sonicate for 20 minutes to obtain a uniformly dispersed nano-gold solution;

[0052] (4) Template molecule tetanus toxin, functional monomer sialyl ganglioside, crosslinking agent trimethylolpropane trimethacrylate, porogen dichloromethane, initiator azobisisobutyronitrile, organic The solvent dichloromethane is mixed uniformly at a molar ratio of 0.5:2.5:3:45:0.1:5 to obtain a tetanus toxin molecularly imprint...

Embodiment 3

[0060] A method for preparing a glycosyl functionalized molecular imprinted column of botulinum toxin, comprising the following steps:

[0061] (1) Select N-acetylglucosamine, a functional monomer capable of synthesizing molecularly imprinted polymers with botulinum toxin;

[0062] (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;

[0063] (3) Take 20 μl of the prepared nano-platinum solution and ultrasonicate for 40 minutes to obtain a uniformly dispersed nano-platinum solution;

[0064] (4) Template molecule bot...

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Abstract

The invention relates to a preparation method of a nano-synergistic carbohydrate functionalized molecularly imprinted column for detecting bacterial toxins, comprising the following steps: selecting a carbohydrate functional monomer which can be used for synthesizing a carbohydrate functionalized molecularly imprinted polymer with the bacterial toxins; preparing solution of the carbohydrate functionalized molecularly imprinted polymer; preparing nano solution; and modifying nano materials and the carbohydrate functionalized molecularly imprinted polymer on the inner surface of a glass tube column. A method for detecting trace bacterial toxins of the invention is characterized by connecting the carbohydrate functionalized molecularly imprinted column prepared by the above method with a chemiluminescence analyzer, respectively pumping chemiluminescence system solution and sample solution into the chemiluminescence analyzer and detecting the bacterial toxins in the samples. The preparation method of the invention has controllability and improves the sensitivity and accuracy of the column. The prepared molecularly imprinted column features strong specificity, high sensitivity and rapid detection speed, can realize high throughout screening of vast samples in short time and lower the detection cost.

Description

technical field [0001] The invention relates to a method for preparing a nano-synergized bacterial toxin glycosyl functionalized molecular imprinted column, and the invention also relates to a method for detecting trace bacterial toxins in a sample by using the glycosyl functionalized molecularly imprinted column, which belongs to trace Bacterial toxin detection technology field. Background technique [0002] 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. With the development of detection methods and analytical techniques, it has been found that bacterial toxins are widely present in almost all food and feedstuffs. Therefore, the development of a method with high accuracy, strong specificity, and rapid...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76
Inventor 黄加栋裴梅山汪世华于京华葛慎光邢宪荣贺晓蕊林青张秀明朱晗孙纳新宋晓妍袁靓
Owner UNIV OF JINAN
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