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Aporate amphion switching biology isolate medium as well as its preparing method and usage

A zwitterion and biological separation technology, applied in the direction of zwitterion exchange, inorganic cation exchanger, inorganic anion exchanger, etc., can solve the problem of difficult separation, achieve good separation reproducibility, fast speed, improve separation and analysis effect of speed

Inactive Publication Date: 2008-04-09
NINGXIA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, most of genetic engineering products are acidic recombinant proteins, and it is difficult to achieve good separation by simply using cation or anion exchange separation media.

Method used

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  • Aporate amphion switching biology isolate medium as well as its preparing method and usage
  • Aporate amphion switching biology isolate medium as well as its preparing method and usage
  • Aporate amphion switching biology isolate medium as well as its preparing method and usage

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1: A kind of non-porous amphoteric ion exchange biological separation medium: its structural formula is:

[0028]

[0029] where the structure of P is:

[0030]

[0031] The preparation method of the non-porous amphoteric ion-exchange bio-separation medium: add 40 mL of 1.0 mol / L dilute HCl to 4.0 g of non-porous monodisperse P (GMA / EDMA) resin, ultrasonically disperse for 10 min, and then react at a constant temperature of 30-40 ° C for 2 h. Suction filtration, repeated washing with distilled water until neutral, add 30mL aqueous solution of dimethylamine to the product, react at constant temperature 60°C for 20h, wash the product repeatedly with a large amount of water and acetone, and vacuum dry to obtain ammoniated microspheres. Add 3g of 1,3-propane sultone to the microspheres in 120mL of acetonitrile, ultrasonically disperse and stir at 80°C for 20 hours at reflux, wash the product with a large amount of acetone and ethanol, and dry it in vacuum. ...

Embodiment 2

[0034] Embodiment 2: A kind of non-porous amphoteric ion exchange biological separation medium: its structural formula is:

[0035]

[0036] where the structure of P is:

[0037]

[0038] The preparation method of the non-porous amphoteric ion-exchange bio-separation medium: add 50 mL of 1.0 mol / L dilute HCl to 5.0 g of non-porous monodisperse P (GMA / EDMA) resin, ultrasonically disperse for 12 min, and react at a constant temperature of 30-40 °C for 2.5 h , filtered with suction, washed repeatedly with distilled water until neutral, then added 40mL of dimethylamine aqueous solution to the product, reacted at a constant temperature of 65°C for 22h, washed the product repeatedly with a large amount of water and acetone, and dried in vacuum to obtain ammoniated microspheres. Add 4g of 1,3-propane sultone into 135mL of acetonitrile, ultrasonically disperse and stir at 85°C for 22 hours at reflux, wash the product with a large amount of acetone and ethanol, and dry it in vacu...

Embodiment 3

[0041] Embodiment three: a kind of non-porous amphoteric ion exchange biological separation medium: its structural formula is:

[0042]

[0043] where the structure of P is:

[0044]

[0045] The preparation method of the non-porous amphoteric ion-exchange bio-separation medium: add 60 mL of 1.0 mol / L dilute HCl to 6.0 g of non-porous monodisperse P (GMA / EDMA) resin, ultrasonically disperse for 15 min, react at a constant temperature of 40 °C for 3 h, and filter with suction , washed repeatedly with distilled water until neutral, then add 50mL of dimethylamine aqueous solution to the product, react at a constant temperature of 70°C for 24 hours, wash the product repeatedly with a large amount of water and acetone, and dry it in vacuum to obtain ammoniated microspheres. The ammoniated microspheres In 150mL of acetonitrile, add 5g of 1,3-propane sultone, reflux and stir at 90°C for 24 hours after ultrasonic dispersion, wash the product with a large amount of acetone and et...

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Abstract

The invention relates to a non-porous amphoteric ion exchange biological separation medium. The structural formula is shown in figure, and the non-porous amphoteric ion exchange biological separation medium has a preparation method that 40 to 60 ML 1.0 mol / L of diluted HCl is added into 4.0 to 6.0 g non-porous monodisperse P (GMA / EDMA) resin, which is reacted for 2 to 3 hours in the constant temperature at 30 to 40 DEG C after being supersonicly dispersed for 10 to 15 minutes, filtered, and repeatedly cleaned with distilled water to be neutral; the water solution of 30 to 50 ML dimethylamine is further added into the resultant, which is reacted for 20 to 24 hours in the constant temperature at 60 to 70 DEG C; the resultant is repeatedly cleaned with a great amount of water and acetone, and performed the vacuum drying, thereby obtaining ammoniation microspheres; the microspheres after ammoniation are laid in acetonitrile of 120 to 150 ML, 3 to 5 g 1,3-Propanesultone is added, the return flow agitation is performed for 20 to 24 hours at 80 to 90 DEG C after the ultrasonic dispersion, and the resultant is cleaned with a great amount of acetone and ethyl alcohol, and performed the vacuum drying, thereby obtaining the non-porous amphoteric ion exchange biological separation medium. The invention takes 3.0 micron non-porous monodisperse porous poly-glycidylmethacrylate-co-ethylene dimethacrylate resin as a matrix, the adoption of a new chemical modified method prepares novel protein which can simultaneously separate the acidity and the alkality, the protein has the advantages of easy regeneration, strong acid and strong base resistance, high mechanical strength, rapid separation velocity, and high protein quality and activity recovery ratio, and the protein can be widely used for rapid separation and purification of gene engineering products and the protein.

Description

technical field [0001] The invention relates to a non-porous amphoteric ion exchange biological separation medium and its preparation method and application. Background technique [0002] Non-porous high-polymer type high-performance liquid chromatography (HPLC) stationary phase has developed rapidly since its appearance in the late 1980s. Compared with porous packing, its most important feature is that it eliminates the peaks caused by stagnant mobile phase mass transfer. Widening improves the column efficiency and separation effect. At the same time, because the separated biomacromolecules do not enter the interior of the particles, they only undergo mass transfer and exchange on the surface, so they can be used for rapid separation and analysis of biomacromolecules such as proteins and peptides. [0003] The amphoteric ion exchange material is a kind of material in which two kinds of exchange groups, yin and yang, exist at the same time in the same resin particle. The tw...

Claims

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Application Information

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IPC IPC(8): B01J43/00B01D15/36
Inventor 龚波林
Owner NINGXIA UNIVERSITY
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