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Method for preparing glycosyl functional bacterial toxin molecularly imprinted column and application thereof

A molecularly imprinted column, bacterial toxin technology, applied in the direction of chemical reaction of materials for analysis, chemiluminescence/bioluminescence, etc., can solve problems such as common cross-reaction, and achieve fast and simple operation, high sensitivity, and avoid subjective factors. effect of influence

Inactive Publication Date: 2011-07-20
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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  • Method for preparing glycosyl functional bacterial toxin molecularly imprinted column and application thereof
  • Method for preparing glycosyl functional bacterial toxin molecularly imprinted column and application thereof
  • Method for preparing glycosyl functional bacterial toxin molecularly imprinted column and application thereof

Examples

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Embodiment 1

[0031] A method for preparing a glycosyl-functionalized diphtheria toxin molecularly imprinted column, comprising the following steps:

[0032] (1) Select the functional monomer A-D-pyranosylmannose that can synthesize glycosyl-functionalized molecularly imprinted polymers with diphtheria toxin;

[0033] (2) template molecule diphtheria toxin, functional monomer, crosslinking agent ethylene glycol dimethacrylate (EGDMA), porogen chloroform, initiator azobisisobutyronitrile, organic solvent dichloromethane in molar ratio 0.1:2.5:0.5:70:0.05:2.0 and mixed uniformly to obtain a glycosyl-functionalized diphtheria toxin molecularly imprinted polymer solution;

[0034] (3) Use a glass column with a length of 5cm and a diameter of 3mm, and use 1mol / L HNO 3 Wash with 1mol / L NaOH, then thoroughly wash several times with double distilled water, and blow dry to ensure that the inner surface of the glass column is bright and free of impurities;

[0035] (4) if figure 1 As indicated, in...

Embodiment 2

[0039] A method for preparing a glycosyl-functionalized tetanus toxin molecularly imprinted column, comprising the following steps:

[0040] (1) Select the functional monomer sialyl ganglioside that can synthesize glycosyl-functionalized molecularly imprinted polymers with tetanus toxin;

[0041] (2) Template molecule tetanus toxin, functional monomer, crosslinking agent trimethylolpropane trimethacrylate, porogen dichloromethane, initiator azobisisobutyronitrile, organic solvent dichloromethane massage Mole ratio 0.5:2.5:3:45:0.1:5, mixed evenly to obtain glycosyl-functionalized tetanus toxin molecularly imprinted polymer solution;

[0042](3) Choose a glass column with a length of 8cm and a diameter of 2mm, and use 1mol / L HNO 3 Wash with 1mol / L NaOH, then thoroughly wash several times with double distilled water, and blow dry to ensure that the inner surface of the glass column is bright and free of impurities;

[0043] (4) if figure 1 As indicated, inject the glycosyl-fu...

Embodiment 3

[0047] A method for preparing a glycosyl-functionalized botulinum toxin molecularly imprinted column, comprising the following steps:

[0048] (1) Select N-acetylglucosamine, a functional monomer capable of synthesizing glycosyl-functionalized molecularly imprinted polymers with botulinum toxin;

[0049] (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 1:2.5:5:65:0.08:15 to obtain a glycosyl-functionalized botulinum toxin molecularly imprinted polymer solution;

[0050] (3) Choose a glass column with a length of 10cm and a diameter of 5mm, and use 1mol / L HNO 3 Wash with 1mol / L NaOH, then thoroughly wash several times with double distilled water, and blow dry to ensure that the inner surface of the glass column is bright and free of impurities;

[0051] (4) The glycosyl-functio...

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Abstract

The invention relates to a method for preparing a glycosyl functional bacterial toxin molecularly imprinted column, which comprises the following steps: selecting a carbohydrate functional monomer which can compound a glycosyl functional molecularly imprinted polymer with bacterial toxin; preparing a glycosyl functional molecularly imprinted polymer solution; and modifying the glycosyl functionalmolecularly imprinted polymer on the inner surface of a glass pipe column by a surface modifying technology. In the method for detecting bacterial toxin, the glycosyl functional molecularly imprintedcolumn prepared by the method is connected to a chemical luminescent analyzer, and a chemical luminescent system solution and the sample solution are respectively pumped into the chemical luminescentanalyzer to detect the bacterial toxin in a sample. The prepared glycosyl functional bacterial toxin molecularly imprinted column has high specificity, sensitivity and detecting sped, can realize high flux screening of a plurality of samples in short time and reduces the detecting cost.

Description

technical field [0001] The invention relates to a preparation method of a glycosyl-functionalized bacterial toxin molecularly imprinted column, and also relates to a method for using the glycosyl-functionalized molecularly imprinted column to detect bacterial toxins in a sample, which belongs to the technical field of bacterial toxin detection. 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 detection of bacterial toxins, rapid...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/76
Inventor 黄加栋于京华汪世华裴梅山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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