Method for preparing lysozyme molecular imprinting nano particles with magnetic responsiveness and extremely high adsorption capacity

A magnetic nanoparticle and molecular imprinting technology, applied in biochemical equipment and methods, chemical instruments and methods, enzymes, etc., can solve the problems of few adsorption sites, large biological protein molecules, and low selectivity of western blotting, etc. Simple, high physical and chemical stability, avoiding cumbersome effects

Inactive Publication Date: 2011-08-10
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The biggest problem is that due to the large size of biological protein molecules, there are fewer adsorption sites constructed, and the selectivity of western blotting is not high.

Method used

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  • Method for preparing lysozyme molecular imprinting nano particles with magnetic responsiveness and extremely high adsorption capacity
  • Method for preparing lysozyme molecular imprinting nano particles with magnetic responsiveness and extremely high adsorption capacity
  • Method for preparing lysozyme molecular imprinting nano particles with magnetic responsiveness and extremely high adsorption capacity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] 1g magnetic material nano-Fe 3 o 4 Mix well with 100mL of 0.7mol / L sodium citrate, react at 40°C for 12 hours, then wash with pure water several times, and then disperse into ethanol solution. Next, add 0.5g of dispersed magnetic fluid, 0.5g of ammonia water and 0.6g of tetraethoxysilane into a mixed solution of 50mL of ethanol and water (5 / 1, V / V), and react for 12 hours at 40°C , coated with SiO on its surface 2 film. Subsequently, disperse 0.05mL of γ-mercaptopropyltrimethoxysilane and 0.05mL of methacryloxypropyltrimethoxysilane into 10mL of pure water, and after reacting for 5 hours, slowly add 200mg of coated SiO 2 thin film Fe 3 o 4 10 mL of pure water, reacted at 60°C for 12 hours, then washed with pure water several times, and dried in vacuum to form a brown powder. Finally, 20 mg of the template molecule lysozyme and acrylamide were dissolved in 10 mL of 0.2 mol / L phosphate buffered saline (pH 5.0) at a molar ratio of 1000:1, sonicated for 10 min, and th...

Embodiment 2

[0051] 15g magnetic material nano-Fe 3 o 4 Mix well with 300mL 0.8mol / L sodium citrate, react at 40°C for 12 hours, then wash with pure water several times, and then disperse into ethanol solution. Next, 5g of dispersed magnetic fluid, 5g of ammonia water and 6g of tetraethoxysilane were added to a mixed solution of 100mL of ethanol and water (2 / 1, V / V), and reacted for 12 hours at 40°C. Surface coated SiO 2film. Subsequently, disperse 0.14mL of γ-mercaptopropyltrimethoxysilane and 0.14mL of methacryloxypropyltrimethoxysilane into 20mL of pure water, and after 5 hours of reaction, slowly drop into the solution containing 600mg of coated SiO 2 thin film Fe 3 o 4 20 mL of pure water in 60°C for 12 hours, then washed several times with pure water and dried in vacuum to form a brown powder. Finally, 40 mg of the template molecule lysozyme and acrylamide were dissolved in 10 mL of 0.4 mol / L phosphate buffered saline (pH 6.0) at a molar ratio of 2000:1, sonicated for 7 minutes...

Embodiment 3

[0053] 50g magnetic material nano-Fe 3 o 4 Mix well with 500mL of 1mol / L oleic acid, react at 40°C for 24 hours, then wash with pure water several times, and then disperse into ethanol solution. Next, 20g of dispersed magnetic fluid, 40g of ammonia water and 12g of tetraethoxysilane were added to a mixed solution of 200mL of ethanol and water (5 / 1, V / V), and reacted for 24 hours at 60°C. Surface coated SiO 2 film. Subsequently, disperse 0.5mL of γ-mercaptopropyltrimethoxysilane and 0.5mL of methacryloxypropyltrimethoxysilane into 50mL of pure water, and after reacting for 10 hours, slowly drop into 2 thin film Fe 3 o 4 50 mL of pure water in 100°C, reacted for 24 hours, then washed with pure water several times, and dried in vacuum to form a brown powder. Finally, 120 mg of the template molecule lysozyme and acrylamide were dissolved in 20 mL of 0.5 mol / L phosphate buffered saline solution (pH 7.0) at a molar ratio of 3000:1, sonicated for 10 min, and then 300 mg of the ...

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Abstract

The invention provides a method for preparing lysozyme molecular imprinting nano particles with magnetic responsiveness and extremely high adsorption capacity. In the method, lysozymes are used as template molecules. The method comprises the following steps of: carrying out template polymerization under the conditions that suitable functional monomers, a cross-linking agent and an initiator and magnetic nano particles the surfaces which are coated with SiO2 and are modified by sulfydryl and acryloyloxy exist; and washing out the template molecules to obtain the lysozyme molecular imprinting nano particles with magnetic responsiveness and high adsorption capacity. The invention also provides a method for preparing the nano particles. The lysozyme molecular imprinting nano particles prepared by adopting the method disclosed by the invention have the characteristics of high adsorption capacity, simpleness for preparation, low cost, strong specific absorption, high physical, chemical stability, capability of being recycled repeatedly and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology and new materials, and relates to the preparation of lysozyme molecularly imprinted nanoparticles, in particular to a preparation method of lysozyme molecularly imprinted nanoparticles with magnetic responsiveness and extremely high adsorption capacity. The specific adsorption and superparamagnetism can realize rapid separation under the action of external magnetism, avoiding the traditional complicated pretreatment procedures. Background technique [0002] Lysozyme, also known as muramidase or N-acetylmuramide glycanohydrlase, is an alkaline enzyme that can hydrolyze mucopolysaccharides in pathogenic bacteria. Mainly by destroying the β-1,4 glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine in the cell wall, the insoluble mucopolysaccharides in the cell wall are decomposed into soluble glycopeptides, resulting in the escape of the contents of the cell wall Bacteria dissolve. Lysoz...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/32B01J20/26C12N9/36B01J20/286
Inventor 梅素容荆涛周宜开戴晴郝巧玲
Owner HUAZHONG UNIV OF SCI & TECH
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