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A preparation method of lysozyme molecularly imprinted temperature-sensitive hydrogel

A molecular imprinting and lysozyme technology, applied in chemical instruments and methods, other chemical processes, etc., can solve the problems of difficult elution and low identification efficiency of molecularly imprinted polymers, and achieve the effect of strong adsorption and improved identification efficiency.

Active Publication Date: 2021-06-04
XIJING UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problems of low recognition efficiency and difficult elution of lysozyme molecularly imprinted polymers, the present invention provides a method for preparing lysozyme molecularly imprinted temperature-sensitive hydrogel, which combines molecularly imprinted technology with temperature-sensitive hydrogel Combined, the obtained hydrogels have high imprinting efficiency and good recognition effect on lysozyme

Method used

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Examples

Experimental program
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Effect test

preparation example Construction

[0026] A method for preparing a lysozyme molecularly imprinted temperature-sensitive hydrogel, comprising the following steps:

[0027] (1) Dissolve 1-carboxymethyl-3-vinylimidazole chloride, NIPAm and Lyz in Tris-HCl buffer solution (pH=7, 10mM, 25°C) according to the mass ratio of 1-5:100:100 , to obtain the prepolymerization system. The mass ratio of Tris-HCl buffer solution to NIPAm is 10:1.

[0028] (2) Add the cross-linking agent N,N-methylenebisacrylamide (BIS) to the prepolymerization system obtained in step (1), and the added amount is 1-5% of the mass of NIPAM.

[0029] (3) Adding initiator ammonium persulfate (APS) to step (2), the amount added is 1-2% of the mass of NIPAm.

[0030] (4) After the mixed solution obtained in step (3) was ultrasonically deoxygenated at 25°C for 20 minutes, nitrogen gas was introduced for 30 minutes, and then tetramethylethylenediamine (TEMED) was quickly added and sealed. The amount of TEMED added is 1-2% of the mass of NIPAm. Reac...

Embodiment 1

[0037] The preparation method of lysozyme molecularly imprinted temperature-sensitive hydrogel comprises the following steps:

[0038] Add 5 mg of 1-carboxymethyl-3-vinylimidazole chloride, 500 mg of NIPAm, and 500 mg of Lyz into 5 g of Tris-HCl buffer solution (pH=7, 10 mM, 25 ° C); then put 5 mg of BIS and 5mg of APS, the resulting mixture solution was deoxygenated by ultrasonic for 20min at 25°C, then nitrogen gas was passed through for 30min, 5mg of TEMED was added quickly, and sealed. After reacting at 25°C for 6h, the Lyz temperature-sensitive hydrogel was obtained.

[0039] The preparation of the blank temperature-sensitive hydrogel does not add Lyz, and the other steps are the same as above.

[0040] The Lyz temperature-sensitive hydrogel and the blank temperature-sensitive hydrogel obtained by the reaction were punched into circular pieces of uniform size with a punch at 25°C, and then put into Tris-HCl buffer containing 1M NaCl respectively. Soak in the solution (p...

Embodiment 2

[0043] The preparation method of lysozyme molecularly imprinted temperature-sensitive hydrogel comprises the following steps:

[0044] Add 10 mg of 1-carboxymethyl-3-vinylimidazole chloride, 500 mg of NIPAm, and 500 mg of Lyz into 5 g of Tris-HCl buffer solution (pH = 7, 10 mM, 25 ° C), followed by 15 mg of BIS and 10mg of APS, the resulting mixture solution was deoxygenated by ultrasonic for 20min at 25°C, then nitrogen gas was passed through for 30min, TEMED 10mg was added quickly, and sealed. After reacting at 25°C for 6h, the Lyz temperature-sensitive hydrogel was obtained.

[0045] The preparation of the blank temperature-sensitive hydrogel does not add Lyz, and the other steps are the same as above.

[0046] The Lyz temperature-sensitive hydrogel and the blank temperature-sensitive hydrogel obtained by the reaction were punched into circular pieces of uniform size with a punch at 25°C, and then put into Tris-HCl buffer containing 1M NaCl respectively. Soak in the solut...

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Abstract

The invention discloses a preparation method of lysozyme molecularly imprinted temperature-sensitive hydrogel, using 1-carboxymethyl-3-vinylimidazole chloride ionic liquid as a functional monomer, N-isopropylacrylamide (NIPAm ) as a supporting monomer, N, N-methylenebisacrylamide (BIS) as a crosslinking agent, ammonium persulfate (APS) and tetramethylethylenediamine (TEMED) as a redox initiator, trimethylol A buffer solution (pH=7, 10mM, 25°C) of aminomethane and hydrochloric acid (Tris-HCl) is used as a solvent, and lysozyme (Lyz) is used as a method for preparing a molecularly imprinted temperature-sensitive hydrogel of an imprinted molecule, and the obtained hydrogel The glue has high imprinting efficiency and good recognition effect for Lyz.

Description

technical field [0001] The invention belongs to the technical field of molecular imprinting, and relates to a preparation method of a lysozyme molecular imprinting temperature-sensitive hydrogel. Background technique [0002] Molecular imprinting technology refers to the process of preparing polymers with specific selectivity for a specific target molecule. The realization of imprinting technology is generally through the pre-assembly of functional monomers and target molecules (ie, imprinted molecules), and then polymerized in the presence of cross-linking agents to obtain polymers that can interact with imprinted molecules. After removal, the remaining polymer has both adsorption and specific holes for the imprinted molecules, which can realize the process of unique recognition of the imprinted molecules. Among them, protein molecular imprinting technology has attracted the attention of many researchers at home and abroad because it can be widely used in chromatographic a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08F220/54C08F226/06C08F222/38C08J9/26B01J20/26B01J20/30
CPCB01J20/268C08F220/54C08J9/26C08J2201/0424C08F226/06C08F222/385
Inventor 高博张煜星宋文琦杨宏伟康小希徐向阳
Owner XIJING UNIV
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