Preparation method of amphiphilic affinity polymer microspheres with core-shell structure

A core-shell structure and polymer technology, applied in the field of amphiphilic affinity microsphere technology and protein protection research, can solve the problems of difficult dispersion, difficult denaturation mechanism, weakened ability of protection and refolding, etc., and achieve the effect of overcoming the difficulty of removal.

Active Publication Date: 2021-12-03
TIANJIN UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This method also encountered some problems at the same time. On the one hand, according to previous literature reports, protein refolding and irreversible denaturation mainly rely on hydrophobic interactions. If the prepared material is too hydrophobic, it will be difficult to disperse in aqueous solution. If the prepared material is too Hydrophilicity leads to a greatly reduced ability to protect and refold
On the other hand, the hydrophobic groups and hydrophilic groups are randomly distributed in the polymer particles, which brings difficulties to the future research on the denaturation mechanism of proteins.

Method used

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  • Preparation method of amphiphilic affinity polymer microspheres with core-shell structure
  • Preparation method of amphiphilic affinity polymer microspheres with core-shell structure

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Embodiment 1

[0037] Embodiment 1 Preparation of an amphiphilic polymer with a core-shell structure

[0038] (1) Preparation of hydrophobic particles poly(divinylbenzene-co-4-vinylphenylboronic acid) microspheres, 10mmol of 4-vinylbenzeneboronic acid, 10mmol of divinylbenzene, dissolved in 160mL of acetonitrile, added 10mg of initiator ammonium persulfate, sealed after passing nitrogen gas. Oil bath at 70°C, polymerize for 30 minutes, after the solution becomes turbid, connect to distillation equipment, increase the temperature of oil bath to 110°C, continue the reaction until half of the volume of acetonitrile is evaporated, and the time is controlled within 2 hours. After the polymerization reaction, use a G5 sand core funnel to carry out suction filtration, then wash with acetone, tetrahydrofuran, and methanol in sequence, and vacuum-dry it for later use.

[0039] (2) Preparation of Amphoteric Core-Shell Polymer Microspheres by Pickering Emulsion Polymerization

[0040] Add 120 mg of t...

Embodiment 2

[0042] Thermostability test of horseradish peroxidase maintained by amphiphilic polymer microspheres with core-shell structure

[0043] Take 5mg of amphoteric core-shell polymer microspheres and add to 2mL to a concentration of 0.5mg mL -1 horseradish peroxidase in aqueous solution. This aqueous horseradish peroxidase solution was heated at 70° C. for 1 hour. After the supernatant was removed by centrifugation, 100 μL of DAB horseradish peroxidase chromogenic reagent was added. The material was observed under an ordinary optical microscope, and it was found that the interior of the microspheres changed from colorless to red, which indicated that the amphiphilic polymer microspheres with the core-shell structure could immobilize the horseradish peroxidase in the interior under high temperature conditions. Hydrophilic core while maintaining the activity of horseradish peroxidase.

[0044] The control group was the above solution without adding amphoteric core-shell polymer mi...

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Abstract

The present invention relates to the preparation of a novel core-shell structure amphiphilic affinity polymer microspheres, comprising the following steps (a) preparing hydrophobic particle poly(divinylbenzene-co-4-vinylphenylboronic acid) by distillation and precipitation method ) microspheres; (b) preparing amphiphilic polymer microspheres with a core-shell structure by means of Pickering emulsion dispersion polymerization. The beneficial effect of the present invention is: (1) the amphoteric nucleophilic polymer of a kind of core-shell structure prepared by the present invention, surface hydrophobic, interior hydrophilic, can be used for under high temperature condition to horseradish peroxidase have better effect. The protection of horseradish peroxidase has better activity and good enzymatic activity. (2) No surfactant is used in the preparation method, which overcomes the problem of difficult removal of traditional surfactants, and will provide an effective means for future research on protein denaturation mechanism.

Description

technical field [0001] The invention belongs to the technology of amphiphilic affinity microspheres with core-shell structure prepared by the Pickering emulsion polymerization method and the research field of protein protection, especially the activity of horseradish peroxidase under high temperature conditions. Background technique [0002] The protein is unstable in vitro and is easily affected by the environment. Under extreme pH conditions, heating, denaturants and organic solvents, the protein structure will be changed, and the hydrophobic groups will be exposed, resulting in the aggregation of protein monomers through hydrophobic interactions. Eventually lead to irreversible denaturation of the protein. Researchers have done a lot of research work on protein protection in vitro, and developed a variety of protein protection reagents, such as surfactants, polymer microspheres, polymeric nanoparticles, polymer micelles, and molecular chaperones. These works have greatly...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08F257/00C08F230/06C08F212/36C08F275/00C12N11/082C12N11/04
CPCC08F257/00C08F275/00C12N9/0065C12N11/04C12N11/08C12Y111/01007C08F220/56
Inventor 王俊平赵涛王硕
Owner TIANJIN UNIV OF SCI & TECH
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