Heparin affinity column and preparation method and use thereof

Abstract

The invention discloses a heparin affinity column as well as the preparation method and the application thereof. The heparin affinity column is prepared through the following method: agarose gel 6FF is taken as a solid phase carrier which is firstly activated and then is aminated and coupled with amidocyanogen after an epoxy group is coupled, the aminated agarose gel 6FF and the heparin form intermediate aldimine in methanol, and the intermediate product of aldimine is further subjected to reductive amination for forming stable chemical bonds, thereby acquiring the heparin-agarose gel 6FF. The affinity column preparing preparation with high efficiency is simple and short. Through adopting the combination of the rear aldehyde of the heparin and the amidocyanogen of aminated sepharose, the activity of the heparin is not influenced, thereby ensuring the affinity column to have higher affinity; the heparin coupling is stable, and the heparin can be repeatedly used; the reagent adopted by the method has low cost and no environmental pollution, thereby laying a foundation for sweepingly isolating and purifying materials with specific binding capacity to the heparin in common laboratories, and promoting the research on domestic relative fields.

Description

technical field [0001] The invention relates to a heparin affinity column and its preparation method and application. technical background [0002] Heparin is a kind of glycosaminoglycan, a mixture of polysaccharide chains composed of repeating disaccharide units connected by uronic acid and glucosamine with 1→4 bonds; it can be used as an effective ligand for affinity chromatography and ion exchange chromatography. It has affinity for many biomolecules, including coagulation factors and other plasma proteins such as lipoproteins, protein synthesis factors, nucleases, and steroid hormone receptors. 2-O-sulfate-α-L iduronic acid and 6-O-sulfate-N-sulfate-α-D glucosamine are the main monosaccharides, which are composed of repeating units of trisulfate disaccharides The so-called "regular region" of heparin is the main part of the heparin structure, and additional monosaccharide residues appear at very low frequency in the "irregular region". In the finished heparin, only one...

AI Technical Summary

Problems solved by technology

As early as 1983 in foreign countries, some people used three methods to link heparin to agarose for comparison. Among the three methods, the amount of heparin consumed by the reductive amination method was the least, but the affinity was the highest; however, the method at that time also had certain defects. That is, the reaction product of the reducing agent sodium cyanoborohydride is highly toxic, pollutes the environment, and the reaction cycle is as long as 16 days.

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