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Preparation method and application of targeted enzyme immobilized carrier based on magnetic molecular imprinting technology

A magnetic molecular imprinting and immobilized carrier technology, which is applied in the field of analysis and detection, can solve the problems of inapplicability, poor resistance of proteins to organic solvents and acid and alkali, difficulty in preparing enzyme molecular imprinting polymers, etc., and achieves low cost and fast preparation process. simple effect

Active Publication Date: 2019-09-17
XI AN JIAOTONG UNIV
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Problems solved by technology

[0004] At present, the imprinted polymer technology of small molecules has become mature, however, for water-soluble protein molecules (such as enzyme molecules), commonly used methods for preparation and recognition in organic phases, such as bulk method, in situ method and precipitation method, etc. , due to the poor resistance of the protein to organic solvents and acid and alkali, it is not applicable, which makes the preparation of enzyme molecularly imprinted polymers difficult.
There is no report on the application of molecularly imprinted polymers to immobilized enzymes

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  • Preparation method and application of targeted enzyme immobilized carrier based on magnetic molecular imprinting technology
  • Preparation method and application of targeted enzyme immobilized carrier based on magnetic molecular imprinting technology
  • Preparation method and application of targeted enzyme immobilized carrier based on magnetic molecular imprinting technology

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Embodiment Construction

[0034] The present invention will be further described in detail below in conjunction with the drawings and embodiments.

[0035] (1) Preparation of horseradish peroxidase magnetic molecularly imprinted polymer

[0036] 1) Amino Fe 3 O 4 Nanoparticles (Fe 3 O 4 -NH 2 ) Preparation

[0037] Weigh FeCl 3 ·6H 2 1.0 g of O, 2.0 g of anhydrous sodium acetate and 6.5 g of 1,6-hexanediamine were sequentially dispersed in 30 mL of ethylene glycol. Stir vigorously at room temperature until the mixture becomes a clear solution. Transfer the above-mentioned transparent solution to a polytetrafluoroethylene lined autoclave, and react at 200°C for 6 hours. Collect the product with a magnet, wash the product with ethanol and water, and then dry it at 50°C to obtain amino Fe with an average particle size of about 20nm 3 O 4 Nanoparticles, spare.

[0038] 2) Boric acid modified Fe 3 O 4 Preparation

[0039] Take the above amino Fe 3 O 4 500 mg of nanoparticles were dispersed in 20 mL of anhydrous me...

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Abstract

The invention discloses a preparation method and application of a targeted enzyme immobilized carrier based on a magnetic molecular imprinting technology. The method comprises the following steps of preparing ferric aminotetroxide; preparing boracic acid modified ferroferric oxide; preparing a molecular imprinting polymer layer by a sol-gel method; removing horseradish peroxidase combined on the molecular imprinting polymer; sealing non-specific adsorption residuals on the surface of the polymers to obtain horseradish peroxidase magnetic molecular imprinting polymers. The method provided by the invention has the advantages that the preparation process is simple; the cost is low; the obtained magnetic molecular imprinting polymers can be used for specifically recognizing the target substance of the horseradish peroxidase, and can also be used for fixing the horseradish peroxidase in complicated samples directly through adsorption; the formed immobilized enzyme can be used for detecting substances such as glucose, sarcosine, trioxypurine and cholesterol capable of being decomposed to produce H2O2.

Description

Technical field [0001] The invention belongs to the field of analysis and detection, and in particular relates to the preparation of a targeted immobilized horseradish peroxidase and a carrier based on magnetic molecular imprinting technology. Background technique [0002] Immobilized enzyme technology is a type of technology that restricts or confines the enzyme to a certain area of ​​the solid support under the condition of maintaining the catalytic activity of the enzyme. Compared with non-immobilized free enzymes, immobilized enzymes maintain their high efficiency, specificity and mild enzyme catalytic reaction characteristics, while overcoming the shortcomings of free enzymes, with high storage stability, easy separation and recovery, and more It has the advantages of repeated use, continuous and controllable operation, and simple process. Immobilized enzymes have been rapidly developed and widely used in disciplines such as chemistry, biology, medicine, food science, and e...

Claims

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Application Information

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IPC IPC(8): C08G77/26C08K9/04C08K3/22C08J9/26C12N9/08C12N11/14G01N33/543
CPCC08G77/26C08J9/26C08K3/22C08K9/04C08K2003/2275C08K2201/01C12N9/0065C12N11/14C12Y111/01007G01N33/543
Inventor 傅强陈国宁舒花王璐王燕常春
Owner XI AN JIAOTONG UNIV
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