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A kind of preparation method of fast-flow temperature-sensitive super-porous biological separation medium

A bio-separation, super-porous technology, applied in chemical instruments and methods, other chemical processes, etc., can solve the problems of poor chemical stability, low processing capacity, low mechanical strength, etc., and achieve simple preparation methods, good biocompatibility, The effect of high mechanical strength

Active Publication Date: 2016-01-13
CHINA UNIV OF PETROLEUM (EAST CHINA)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it will take time for temperature-sensitive separation media to be used as an effective industrial separation method. It is used in analysis mode, and there are few reports on the separation of biological macromolecules such as proteins; 2) The matrix of temperature-sensitive media in the literature is mainly silica gel and polysaccharide soft gel
The former has poor chemical stability, is not resistant to acid and alkali, and is difficult to clean in industrial applications; the latter has low mechanical strength and can only be used under low pressure, with slow separation speed and difficult industrial scale-up

Method used

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  • A kind of preparation method of fast-flow temperature-sensitive super-porous biological separation medium
  • A kind of preparation method of fast-flow temperature-sensitive super-porous biological separation medium
  • A kind of preparation method of fast-flow temperature-sensitive super-porous biological separation medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Take 2g of super-porous polystyrene microspheres (PS) and add 50mlCS 2 Soak, followed by adding anhydrous AlCl 3 3.7g and 3.02g of chloroacetyl chloride were stirred and reacted at 40°C for 3h, and the product was washed with absolute ethanol, 3% HCl and deionized water successively, and dried in vacuum at 25°C for 3h to prepare PS-Cl.

[0039] Weigh 0.25g of PS-Cl and place it in a pear-shaped Schlenk bottle 1, fill it with argon gas for deoxygenation three times, and set aside; then put the catalyst CuCl (20.54mg) ligand PMDETA (143.54mg) and functional monomer MEO2MA (2.164g) Mix well with OEGMA (0.475g) and solvent methanol / water (4:1, 15ml) in the pear-shaped Schlenk bottle 2, freeze with liquid nitrogen and evacuate the oxygen three times; Introduce into the pear-shaped Schlenk bottle 1; finally, transfer the pear-shaped Schlenk bottle 1 to a water bath shaker for 18 hours of water bath shaking reaction (rotation speed: 140r / min), and the water temperature is 30°...

Embodiment 2

[0041] Take 2g of ultra-large-porous polystyrene microspheres (PS) and soak them in 30ml of dichloromethane, then add anhydrous AlCl 3 4.2g and 3.89g of 2-chloroisobutyryl chloride were reacted with stirring at 50°C for 5h. The product was washed with absolute ethanol, 3% HCl and deionized water successively, and dried in vacuum at 40°C for 2h to prepare PS-Cl.

[0042] Weigh 0.25g PS-Cl and place it in a pear-shaped Schlenk1 bottle, fill it with argon to deoxygenate three times, and set aside; then put the catalyst CuCl (20.46mg), the ligand Me 6 TREN (190.5mg)﹑functional monomer MEO 2 MA (3.576g) and OEGMA (0.475g) ﹑ solvent isopropanol / water (4:1, 17ml) were placed in a pear-shaped Schlenk bottle 2 and mixed evenly, and liquid nitrogen was frozen and evacuated for three times to remove oxygen; The liquid in the pear-shaped Schlenk bottle 2 was introduced into the pear-shaped Schlenk bottle 1; finally, the pear-shaped Schlenk bottle 1 was transferred to a water bath shaker ...

Embodiment 3

[0044] Take 2.5g of ultra-large-porous polystyrene microspheres (PS) and soak them in 35ml of dichloroethane, then add anhydrous AlCl 3 4.4g and 4.52g of bromoacetyl bromide were stirred and reacted at 50°C for 5h. The product was washed with absolute ethanol, 3% HCl, and deionized water in turn, and dried in vacuum at 35°C for 3h to prepare PS-Br.

[0045] Weigh 0.26gPS-Br and place it in the pear-shaped Schlenk bottle 1, fill it with argon to remove oxygen three times, and set it aside for use; then put the catalyst CuCl (18.3mg) / CuCl 2 (3.0mg), ligand Bipy (130.73mg), functional monomer N-isopropylacrylamide (NIPAM) / butyl methacrylate (BMA) (90:10, 2.46g), solvent methanol / water (3 : 1, 22ml) in the pear-shaped Schlenk bottle 2 and mixed evenly, liquid nitrogen freezing and vacuum deoxygenation three times; then use double needles to introduce the liquid in the pear-shaped Schlenk bottle 2 into the pear-shaped Schlenk bottle 1; Bottle 1 was transferred to a water bath shak...

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Abstract

The invention relates to the field of functional polymer material and bio-separation and in particular relates to a preparation method of a thermosensitive super-macroporous bio-separation medium with high flow velocity. The preparation method comprises the steps of firstly introducing an atom transfer radical polymerization (ATRP) initiator to the surface of a super-macroporous polystyrene (PS) microsphere through Friedel-Crafts reaction, and then grafting a thermosensitive polymer brush to the surface of the super-macroporous PS microsphere by utilizing heterogeneous ATRP, thus obtaining the thermosensitive super-macroporous bio-separation medium. The thermosensitive super-macroporous bio-separation medium with both the hydrophobic effect and the ion exchange effect can be prepared by selecting and mixing a hydrophobic monomer, a pH sensitive monomer and a thermosensitive monomer. The thermosensitive polymer brush grafted under the optimized reaction conditions not only has good thermo-responsivity but also can maintain the super-macroporous structure of the PS microsphere. The preliminary separation experiment results show that the thermosensitive super-macroporous bio-separation medium can separate two protein mixtures at 1806cm / h only by changing the temperature, which indicates that the prepared thermosensitive super-macroporous bio-separation medium with high flow velocity has potential in the field of large-scale protein separation and purification.

Description

technical field [0001] The invention belongs to the field of functional polymer materials and bio-separation, in particular to a fast-flow temperature-sensitive super-macroporous bio-separation medium and a preparation method thereof. Background technique [0002] Biomolecules such as peptides and proteins are currently separated by one or a combination of reversed-phase chromatography, ion-exchange chromatography, and hydrophobic interaction chromatography. Among them, reversed-phase chromatography is one of the most widely used chromatographic techniques in the separation, purification and research of peptides. However, the use of organic solvents may lead to the loss of peptide biological activity and further application. Although there is no addition of organic solvents in hydrophobic chromatography, the hydrophobic interaction between proteins and peptides and the stationary phase is promoted by the "salting out" effect caused by the high salt concentration in the mobil...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08F257/02C08F12/08C08F8/24C08F8/18B01J20/285B01J20/30
Inventor 曲剑波陈艳丽宦关生张晓云黄方
Owner CHINA UNIV OF PETROLEUM (EAST CHINA)
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