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Method for preparing double-channel hydrophilic bicontinuous polymer monolithic column

A polymer and hydrophilic technology, applied in the field of dual-channel hydrophilic bicontinuous polymer monolithic column and its preparation, can solve the problems of poor mechanical strength, low separation efficiency, poor biocompatibility, etc., to ensure uniformity Effect

Active Publication Date: 2019-04-19
CHINA UNIV OF PETROLEUM (EAST CHINA)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Second is the issue of biocompatibility of monolithic columns
At present, there are three main types of polymer monolithic columns used for separating biomacromolecules: polystyrene, polyacrylamide and poly(meth)acrylate, but they all have certain defects: polystyrene monolithic columns have good Mechanical strength and chemical stability, but the lack of functional groups on the surface can only be used as a reversed-phase mode. The use of an organic solvent on the surface of a strong hydrophobic medium will lead to protein denaturation and inactivation. If you want to apply it, you can only use hydrophilic modification (Macromolecules 2001, 34, 4361–4369; ACS Appl. Mater. Interfaces, 2014, 6, 12752-12760)
[0006] In summary, the application of polymer monolithic columns in the separation of biological macromolecules still has problems such as low separation efficiency, poor mechanical strength, and poor biocompatibility. It is still a challenge to prepare an ideal polymer monolithic column to overcome the above problems.

Method used

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  • Method for preparing double-channel hydrophilic bicontinuous polymer monolithic column
  • Method for preparing double-channel hydrophilic bicontinuous polymer monolithic column
  • Method for preparing double-channel hydrophilic bicontinuous polymer monolithic column

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] 1) Synthesis of poly 3-O-methacryloyl-diacetone-D-glucose (PMDAGlu) by ATRP reaction

[0069] Add a stirring bar to the Schlenk bottle at room temperature, then add cuprous bromide (5.9mg), N,N,N,N',N'-pentamethyldiethylenetriamine (PMDETA) (7.1mg ), MDAGlu (0.98g) and 3ml chlorobenzene, through three cycles of liquid nitrogen freezing-pumping-gassing-thawing to remove oxygen. Finally, the initiator 2-bromoisobutyrate (30.4 mg) was added, reacted at 50° C. for 4 h, dissolved the reaction product with tetrahydrofuran, diluted the reaction product, and removed the catalyst through an aluminum oxide column. The obtained colorless solution was precipitated twice with n-hexane, suction filtered at room temperature, and dried in vacuo to obtain 0.6 g of PMDAGlu, whose molecular weight Mn=4700 was detected by gel permeation chromatography (GPC).

[0070] 2) Synthesis of poly 3-O-methacryl-diacetone-D-glucose-polystyrene block copolymer (PMDAGlu-PS) by ATRP reaction

[0071] ...

Embodiment 2

[0075] 1) Synthesis of 3-O-acryloyl-diacetone-D-glucose (PDAGlu) by RAFT polymerization

[0076] Add a stirring bar to a 5mL chicken heart bottle at room temperature, then add sugar-containing monomer DAGlu (0.7g), chain transfer agent CPADB (30mg), initiator AIBN (2mg) and solvent dioxane (1mL) in sequence, dissolve and mix After uniformity, use argon purging to remove oxygen in the system. Then react at 75°C for 15 hours, dilute the reaction solution with tetrahydrofuran, precipitate with n-hexane two to three times, and use centrifugation to obtain a solid product, dry it in vacuum at room temperature to obtain 0.37g PDAGlu, and determine the molecular weight M by GPC. n =3450.

[0077] 2) Synthesis of PDAGlu-PS by RAFT polymerization

[0078] Add and stir once in the 10mL heart bottle at room temperature, then add one block polymer PDAGlu (0.3g), initiator AIBN (2.8mg), styrene (3.2g) and solvent dioxane (3mL) successively, use Argon was purged to remove oxygen, and fin...

Embodiment 3

[0084] 1) Synthesis of 3-O-acryloyl-diacetone-D-galactose (PADAGlu) by ATRP reaction

[0085] Add a stirring bar to the Schlenk bottle at room temperature, then add cuprous bromide (5.48mg), N,N,N,N',N'-pentamethyldiethylenetriamine (PMDETA) (7.65mg ), 3-O-methacryloyl-diacetone-D-galactose (MDAGal) (1.30g) and 3.5ml toluene, through three cycles of liquid nitrogen freezing-pumping-gassing-thawing to remove oxygen. Finally, the initiator 1-bromoethylbenzene (18.3 mg) was added, reacted at 60° C. for 4 h, dissolved the reaction product with chloroform, diluted the reaction product, and removed the catalyst through an aluminum oxide column. The resulting colorless solution was precipitated twice with methanol, suction filtered at room temperature, and dried in vacuo to obtain 0.69 g of PMDAGal, whose molecular weight Mn=6800 was detected by gel permeation chromatography (GPC).

[0086] 2) Use ATRP reaction to synthesize 3-O-acryloyl-diacetone-D-glucose-polystyrene block copolym...

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Abstract

The invention relates to the field of polymer monolithic column preparation, in particular to a double-channel hydrophilic bicontinuous polymer monolithic column and a preparation method thereof. A self-made amphiphilic diblock sugar-containing polymer is used as a surfactant to control a continuous phase emulsion, and the block polymer is also involved in the preparation of an initiator or a chain transfer agent for the polymerization of the monolithic column. With the combination of a small molecular pore-forming agent and through one-step polymerization of an active emulsion, the double-channel hydrophilic bicontinuous polymer monolithic column is obtained. According to the prepared double-channel hydrophilic bicontinuous polymer monolithic column, the skeleton of the monolithic columnis polystyrene, which has high mechanical strength and good chemical stability, and the outer surface of the skeleton is a sugar-containing polymer and has hydrophilicity and is easy to derive. The monolithic column pores penetrate, have two pores of super macropores and mesopores, and have wide application space in the fields of cell culture, enzyme immobilization and rapid separation and purification of proteins.

Description

technical field [0001] The invention relates to the field of preparation of polymer monolithic columns, in particular to a dual-channel hydrophilic bicontinuous polymer monolithic column and a preparation method thereof. Background technique [0002] With the rapid development of modern biotechnology, the research and utilization of biomacromolecules such as proteins, virus vaccines, and plasmids has become more and more extensive. However, most of these biomacromolecules have complex spatial structures and are easy to inactivate. Efficient separation has become a research hotspot. In order to reduce the activity loss of biomacromolecules during the separation process, the separation time should be shortened as much as possible under the premise of ensuring the separation purity, so as to reduce the degree of biomacromolecules being cut by the fluid. At the same time, the pore size of the separation medium is generally required to be larger than that of solute molecules. 10...

Claims

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Application Information

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IPC IPC(8): C08F287/00C08F212/08C08F212/36C08F293/00C08J9/26B01J20/281
CPCB01J20/281C08F287/00C08F293/00C08J9/26C08F212/08C08F212/36
Inventor 曲剑波刘源李士海蔺洋洋彭文舒张晓云黄方
Owner CHINA UNIV OF PETROLEUM (EAST CHINA)
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