Affinity chromatography purification method of placenta sample chondroitin sulfate A or derivative thereof

Abstract

The invention relates to an affinity chromatography purification method of placenta sample chondroitin sulfate A or a derivative thereof, and particularly discloses a purification method of the placenta sample chondroitin sulfate A or the derivative thereof. The placenta sample chondroitin sulfate A or the derivative thereof are chromatographically purified by the affinity chromatography method, and an affinity chromatography column is a conjugate of a recombinant plasmodium infected red cell surface antigen protein and an affinity chromatography substrate. According to the chromatography purification method, crude products of the placenta sample chondroitin sulfate A or the derivative thereof are loaded onto the affinity chromatography column and washed by washing liquid until no impurities flow out, and then the crude products are eluted by eluent, and pure products of the placenta sample chondroitin sulfate A or the derivative thereof are collected. The purification method can solvethe problem of difficulty in polysaccharide purification, other polysaccharides with similar molecular sizes and structures cannot be adsorbed by rVAR2 and can be easily washed and removed to obtainhigh-purity pl-CSA, and the washing liquid pl-CSA is firmly combined by an rVAR2 ligand and easily released to obtain high recovery rate.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to an affinity chromatography purification method for placenta-like chondroitin sulfate A or derivatives thereof. Background technique [0002] The principle of affinity chromatography is to use the specificity and reversibility of the affinity between biomolecules. The coupled affinity ligand is based on the stationary phase adsorption medium. The impurities are washed out, and finally by changing the conditions of the mobile phase, such as pH value or ion concentration and type, the affinity adsorption between the dissociated ligand and the target is released to release the target into the mobile phase, thereby obtaining a high-purity target. Since the emergence of affinity chromatography technology, this technology has developed very rapidly and has achieved remarkable results in the field of biotechnology. It has been widely used in proteins, peptides, enzyme preparations, antigen ant...

AI Technical Summary

Problems solved by technology

However, there are still few methods for the establishment and application of polysaccharide affinity purification. The key difficulty lies in the discovery of ligands that can specifically bind to polysaccharides.

[0003] The polysaccharide extraction scheme usually adopts methods such as alcohol extraction, water extraction, or organic solvent extraction to separate the total polysaccharide or polysaccharides of a certain type, such as using Sevag solution (V 氯仿 :V 正丁醇 =4:1) added to the polysaccharide-containing sample, vigorously shaken for 5 minutes and then centrifuged at 4000rpm for 10 minutes, using the interaction of protein with chloroform and n-butanol to form a gel insoluble matter, while the polysaccharide is dissolved in the water phase, and the operation is repeated many times to achieve Remove impurities such as protein, but the polysaccharide obtained by this method is a crude product, containing a variety of polysaccharide components, the quality of subsequent research, development and application is uncontrollable, the type of polysaccharide is uncontrollable, and it is difficult to distinguish the properties of different polysaccharides , type of action and mode of action, such as the residue of other polysaccharides in heparin seriously hinders its clinical application

In order to purify a certain monomer from crude polysaccharides, a more mature method is to use molecular sieve chromatography columns, such as DEAE-cellulose-52 chromatography columns, Sephadex G-100 chromatography columns and AB resins, etc., this method The purification of polysaccharides has been widely used, such as isatis root polysaccharide purification, heparin purification, spirulina polysaccharide purification, etc. This method has a wide range of advantages before there is no better method, but its disadvantage is that the purity is still low and the recovery rate is low , in order to improve the purity, the recovery rate of more than 50% is often sacrificed, and there is nothing to do for substances with similar molecular size and structure

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