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Epoxy activated affinity chromatography medium

A chromatography medium and epoxy-activated technology, applied in the field of epoxy-activated affinity chromatography medium, can solve problems such as reduction in the number of ligand coupling, matrix damage, and reduced efficiency of affinity chromatography, achieving a large number, The density of epoxy groups is high, which is conducive to the effect of large-scale popularization and application

Active Publication Date: 2018-04-03
SUZHOU BOJIN BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Usually, agarose gel is used as the matrix, and epichlorohydrin or allyl glycidyl ether is used to activate the agarose gel, and then it can be directly used for coupling of various ligands, but the highly alkaline activation conditions are easy to cause matrix At the same time, the epoxy group will cross-link with the hydroxyl group on the matrix in a strong alkaline environment, reducing the number of coupling ligands and reducing the efficiency of affinity chromatography

Method used

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  • Epoxy activated affinity chromatography medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The preparation process is as follows:

[0021] (1) Soak 20 g of cellulose microspheres with a particle size of 30-100 μm in 100 mL of sulfuric acid solution with a concentration of 50 wt %, stir magnetically at 55-60° C. for 30 minutes, and filter to obtain activated cellulose microspheres;

[0022] (2) Mix 25g of 1,2-epoxydodecane, 10g of polylactic acid-glycolic acid copolymer, 20g of polyethylene glycol, and 0.2g of diethylenetriamine, and add 50mL of absolute ethanol. Stir at ℃ for 30 minutes and sonicate for 15 minutes to form a homogeneous liquid complex;

[0023] (3) Place the activated cellulose microspheres obtained in step (1) in the liquid compound of step (2), stir at 55-60°C for 3-4 hours, lower the temperature and stir at 45-50°C for 2- 3 hours, then lower the temperature to 20-25°C and stir for 1-2 hours, and finally stir at 5-10°C for 1-2 hours to end the reaction, filter the microspheres, rinse with deionized water and dry to obtain epoxy-activated af...

Embodiment 2

[0025] The preparation process is as follows:

[0026] In the step (2) of Example 1, 1,2-epoxydodecane was replaced with 35 g of 1,2-epoxyoctadecane, the amounts of the two substances were the same, and the rest of the preparation process was the same as in Example 1.

Embodiment 3

[0028] The preparation process is as follows:

[0029] In step (2) of Example 1, 1,2-epoxydodecane was replaced with 12g of 1,2-epoxyhexadecane and 23g of 1,2-epoxypentadecane, and the rest of the preparation process was the same as in Example 1.

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Abstract

The invention relates to an epoxy activated affinity chromatography medium. Cellulose microspheres are taken as a matrix, a compound comprising 1,2-epoxy alkane, polylactic-co-glycolic acid and polyethylene glycol is crosslinked on the surfaces of the cellulose microspheres, and 1,2-epoxy alkane is linked with ligand, wherein 1,2-epoxy alkane comprises one or more of 1,2-epoxy dodecane, 1,2-epoxytridecane, 1,2-epoxy tetradecane, 1,2-epoxy pentadecane, 1,2-epoxy hexadecane, 1,2-epoxy heptadecane and 1,2-epoxy octodecane. The affinity chromatography medium has high epoxy density on the surface,contains numerous coupling ligands and has high affinity chromatography efficiency; meanwhile, the preparation process is simple and favorable for large-scale popularization and application.

Description

technical field [0001] The invention relates to a chromatographic medium, in particular to an epoxy-activated affinity chromatographic medium. Background technique [0002] Affinity chromatography technology is based on the principle of affinity adsorption between biomolecules and other ligand molecules (such as antigens and antibodies, enzymes and substrates, hormones and receptors, complementary chains in nucleic acids, polysaccharides and protein complexes, etc.) established and developed. Affinity chromatography achieves the purpose of purifying biological macromolecules such as target proteins through specific adsorption and dissociation between ligands on the medium and target molecules. Due to this specific force, affinity chromatography has high selectivity It is extremely effective for separating biologically active substances with very small content and unstable content, and has become an important method for separating and purifying biologically active substances...

Claims

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Application Information

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IPC IPC(8): B01J20/26B01J20/28B01J20/30B01D15/38C07K1/22
CPCB01D15/3804B01J20/24B01J20/267B01J20/28011C07K1/22
Inventor 瞿欢欢朱至放
Owner SUZHOU BOJIN BIOLOGICAL TECH
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