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

A technology of magnetic molecular imprinting and immobilization carrier, 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 preparation of enzyme molecularly imprinted polymers, etc., and achieves simple preparation process and low cost. low cost effect

Active Publication Date: 2021-01-19
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 targeting enzyme immobilized carrier based on magnetic molecular imprinting technology
  • Preparation method and application of targeting enzyme immobilized carrier based on magnetic molecular imprinting technology
  • Preparation method and application of targeting enzyme immobilized carrier based on magnetic molecular imprinting technology

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Embodiment Construction

[0034] The present invention will be described in further detail below in conjunction with the accompanying 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 O 1.0g, 2.0g of anhydrous sodium acetate and 6.5g of 1,6-hexanediamine were dispersed in 30mL of ethylene glycol in sequence. Stir vigorously at room temperature until the mixture becomes a clear solution. The above transparent solution was transferred to a polytetrafluoroethylene-lined autoclave and reacted at 200 °C for 6 h. Collect the product with a magnet, wash the resulting product with ethanol and water, and then dry it at 50 °C to obtain amino Fe with an average particle size of about 20 nm 3 o 4 Nanoparticles, spare.

[0038] 2) Boronic acid modified Fe 3 o 4 preparation of

[0039] Take the above amino Fe 3 o 4 500 mg of nanoparticle...

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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 targeted immobilized horseradish peroxidase and its carrier based on magnetic molecular imprinting technology. Background technique [0002] Immobilized enzyme technology is a type of technology that restricts or binds enzymes to a certain area of ​​a solid phase carrier under the condition of maintaining the catalytic activity of the enzyme. Compared with unimmobilized free enzymes, immobilized enzymes overcome the shortcomings of free enzymes while maintaining their high-efficiency, specificity and mild enzymatic reaction characteristics. It has the advantages of repeated use, continuous and controllable operation, and simple process. Immobilized enzymes have been rapidly developed and widely used in chemistry, biology, medicine, food science, and environmental science. The preparation methods of immobilized enzymes can be divided into two ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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