Chromatography medium adopting tryptamine as functional ligand

A chromatographic medium and matrix technology, applied in the field of protein chromatographic separation technology, can solve the problems of weak salt-tolerant binding ability of ligands, affecting antibody activity, etc. Effect

Inactive Publication Date: 2018-05-08
SUZHOU BOJIN BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In practical applications, some peptide ligands interact strongly with antibodies and need to be eluted under strong acidic or alkaline co

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Take 10 g of polyglycidyl methacrylate chromatography matrix with cellulose molecules bonded on the surface, add 10 g of 25% by volume dimethyl sulfoxide, 5 g of bromoacetyl bromide and 6 g of sodium hydroxide, and shake at 200 rpm at 30 ° C. Activate in the bed for 15 hours, filter with suction, wash with deionized water to obtain an activated chromatography matrix; mix the activated chromatography matrix with 3g of N-bromosuccinimide for bromoalcoholization, and react in a shaker at 200rpm at 30°C For 2 hours, filter with suction and wash with deionized water to obtain a bromoalcoholated chromatographic matrix; mix the bromoalcoholated chromatographic matrix with 10 g of tryptamine and 0.5M sodium carbonate buffer, and the pH of the sodium carbonate buffer is 10 , reacted in a shaker at 200 rpm at 25°C for 10 hours to obtain arginine-coupled medium; the tryptamine-coupled medium was sequentially washed with N-methylpyrrolidone, absolute ethanol and deionized water, and...

Embodiment 2

[0023] Take 10 g of polyglycidyl methacrylate chromatography matrix with cellulose molecules bonded on the surface, add 5 g of 25% by volume dimethyl sulfoxide, 8 g of bromoacetyl bromide and 6 g of sodium hydroxide, and shake at 200 rpm at 30 ° C. Activate in the bed for 25 hours, filter with suction, wash with deionized water to obtain the activated chromatography matrix; mix the activated chromatography matrix with 3g N-bromosuccinimide for bromoalcoholization, and react in a shaker at 200rpm at 30°C For 2 hours, filter with suction and wash with deionized water to obtain a bromoalcoholized chromatographic matrix; mix the bromoalcoholated chromatographic matrix with 10 g of tryptamine and 0.8M sodium carbonate buffer, and the pH of the sodium carbonate buffer is 12 , and reacted in a 200rpm shaker at 25°C for 17 hours to obtain arginine-coupled medium; the tryptamine-coupled medium was sequentially washed with N-methylpyrrolidone, absolute ethanol and deionized water, and su...

Embodiment 3

[0025] Take 10 g of polyglycidyl methacrylate chromatography matrix with cellulose molecules bonded on the surface, add 6 g of 25% by volume dimethyl sulfoxide, 8 g of bromoacetyl bromide and 7 g of sodium hydroxide, and shake at 200 rpm at 30 ° C. Activate in the bed for 26 hours, filter with suction, wash with deionized water to obtain the activated chromatography matrix; mix the activated chromatography matrix with 6g N-bromosuccinimide for bromoalcoholization, and react in a shaker at 200rpm at 30°C 2 hours, suction filtration, wash with deionized water, obtain the chromatographic matrix of bromoalcoholization; The chromatographic matrix of bromoalcoholation is mixed with 10g tryptamine and 0.5-1M sodium carbonate buffer solution, the pH of sodium carbonate buffer solution is 12. React in a 200rpm shaker at 25°C for 20 hours to obtain the arginine-coupled medium; wash the tryptamine-coupled medium with N-methylpyrrolidone, absolute ethanol and deionized water in sequence, a...

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Abstract

The invention relates to a chromatography medium adopting tryptamine as a functional ligand. The chromatography medium comprises a chromatography matrix and a ligand, wherein the chromatography matrixis hydrophilic porous microspheres, and the ligand is trytamine coupled after being activated by bromoacetyl bromide. The chromatography medium is high in antibody adsorption capacity, the saturatedadsorption capacity can reach 140mg/ml of wet medium, and the processing capacity is high; and the medium is stable in characteristics, and the ligand is activated and coupled by adopting the allyl bromide, so that the ligand of the medium is stable, and convenience in cleaning and regeneration can be realized.

Description

technical field [0001] The invention relates to a chromatographic medium with tryptamine as a functional ligand, which belongs to the protein chromatographic separation technology in the field of biochemical industry. Background technique [0002] Affinity chromatography is a chromatographic technique that uses the specific affinity between biomolecules to separate them. It has long been recognized that biological macromolecules such as proteins and enzymes can specifically and reversibly bind to some corresponding molecules, and can be used for the separation and purification of biological molecules. Biomolecular pairs with specific affinity mainly include: antigen and antibody, DNA and complementary DNA or RNA, enzyme and its substrate or competitive inhibitor, hormone (or drug) and their receptor, vitamin and its specificity Binding proteins, glycoproteins and their corresponding plant lectins, etc. The carrier used in affinity chromatography is called a matrix, and the...

Claims

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Application Information

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IPC IPC(8): B01J20/26B01J20/28B01J20/30B01D15/38
CPCB01D15/3885B01J20/261B01J20/28021B01J20/28054
Inventor 瞿欢欢朱至放
Owner SUZHOU BOJIN BIOLOGICAL TECH
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