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Monolithic columns combining deep eutectic solvent monomers and hybrid monomers

A deep eutectic solvent and monolithic column technology, applied in the field of separation and analysis, can solve the problems affecting the life and durability of the monolithic column, and achieve the effects of easy control of reaction conditions, good mechanical strength, and high enrichment effect

Active Publication Date: 2021-11-19
TIANJIN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no literature report on the use of DES monomer for the preparation of monolithic columns. During the experiment, we found that the monolithic columns prepared with DES monomers are relatively soft, which affects the life and durability of monolithic columns.

Method used

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  • Monolithic columns combining deep eutectic solvent monomers and hybrid monomers
  • Monolithic columns combining deep eutectic solvent monomers and hybrid monomers
  • Monolithic columns combining deep eutectic solvent monomers and hybrid monomers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Preparation of functional monomers,

[0025] Preparation 1.DES monomers: the choline chloride (60 mmol, 8.4 g) and methacrylic acid (120 mmol, 10.2mL) in a 1: 2 molar ratio threaded vial was added, followed by 90 deg.] C oil bath for 1 H , a uniform transparent solution can be obtained. Placed in a desiccator;

[0026] 2. Preparation of hybrid monomers: Take 1498 μL (6.4 mmol) of 3-aminopropyl triethoxysilane (the APTES) and 688 μL (8.1 mmol) of methacrylic acid (MAA) into the reaction flask mixture ultrasound 10 min dissolution. And then placed at 60 ℃ pot, removed 24 h after the reaction, to give a clear yellow liquid is the hybrid monomer APTES-MAA;

Embodiment ２

[0028] Preparation of monolithic column

[0029] In 670 μL of methanol and 70 mg PEG 2000 was added two yuan porogen functional monomer APTES-MAA (64 μL) and DES monomer ChCl-MAA (144 μL), a crosslinking agent ethylene glycol dimethacrylate ( 62 μL) and the initiator azobisisobutyronitrile (2.6 mg), after the formation of the ultrasonic mixing was poured into the prepolymerization already processed capillary, capillary ends sealed with rubber plugs, placed in the pot 60 deg.] C water bath, after 2 h the reaction capillary removed, rinsed, and other substances soluble matter monolith unreacted with acetonitrile.

Embodiment 3

[0031] To determine the effect of the type of functional monomer extraction efficiency of protein synthesis a single monomer (APTES-MAA monolith, DES monolith) and a conventional monomer (MAA monolithic column) is used as a control monolith. Specific steps are as follows:

[0032] DES / APTES-MAA prepared monolithic column prepared in Example 2 is the monolithic columns embodiment;

[0033] Control group was prepared monolithic column

[0034] Preparation APTES-MAA Monolith: DES except that no monomer was added, the same methods, and experimental conditions in Example 2 Synthesis embodiment; DES monolith prepared: except that no monomer was added APTES-MAA, the same as in Example 2, synthetic methods, and experimental conditions; MAA monolith prepared: except that no monomer was added and DES APTES-MAA monomer, the monomer MAA was added 210 L, synthesized by the same method of Example 2 and the experimental conditions. After completion of the polymerization, rinsed with acetonitri...

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Abstract

The present invention relates to a monolithic column combining deep eutectic solvent monomers and hybrid monomers. Using DES monomer (choline chloride-methacrylic acid) and hybrid monomer (3-aminopropyltriethoxysilane-methacrylic acid) as binary functional monomers, ethylene glycol dimethacrylate As the cross-linking agent, methanol and PEG 20000 were used as binary porogens, and a new type of monolithic column was prepared by in-situ thermal polymerization technology. The invention has the advantages of low cost and simple synthesis process, and the novel monolithic column produced has good permeability, strong mechanical strength and biocompatibility, and can successfully separate bovine serum albumin and cytochrome C. Proteins can be better enriched. The proposed method provides a new idea for the separation and purification of proteins.

Description

Technical field [0001] The present invention belongs to the field of separation and analysis technology, particularly relates to a novel combination of monomers and solvent eutectic monomers hybrid monolith. Background technique [0002] Protein is a kind of biological macromolecules of biological activity, and various life activities are closely related. With the development of proteomics and the Human Genome Project, people want to understand cell biology at the molecular level. Role of protein biochemistry and biological functions attempt to reveal gene product. While maintaining biological activity and chemical integrity of the protein, the desired protein isolated from complex matrices, for the development of life sciences, molecular biology is of great significance. Complete protein separation technologies include polyacrylamide gel electrophoresis, capillary electrophoresis and HPLC. However, the use of particle packing column chromatography for protein separation is chall...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/26B01J20/28B01J20/30B01D15/22G01N23/2251G01N27/447G01N33/68C07K14/765C07K1/14
CPCB01D15/22B01J20/261B01J20/28014B01J20/28042B01J20/3092B01J2220/4812C07K1/14C07K14/765G01N23/2251G01N27/44717G01N33/6803G01N2550/00
Inventor 刘照胜柴美红黄艳萍
Owner TIANJIN MEDICAL UNIV
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