Composite continuous bed cryogel and preparation thereof, and application in separating IgG and albumin

Abstract

The invention discloses a composite continuous bed cryogel and a preparation method thereof, and application in quickly separating human serum IgG and albumin. The pore size of the composite continuous bed cryogel is 0.1-300 mu m, and the porosity is 80-95%; and the composite continuous bed cryogel has a hydrophobic benzyl-anion exchange tertiary amino functional group disclosed as Formula (I). The composite continuous bed cryogel polymer chain simultaneously contains amino ion-exchange group and benzyl group with certain hydrophobic functions, so the cryogel can form multi-point adsorption with different competitivenesses with the IgG macromolecule and albumin; and thus, after the composite continuous bed cryogel is sequentially decomposed after being eluted by saliferous eluates with different concentrations. The composite continuous bed cryogel has the advantages of high separation purity and favorable separating properties. The chromatography method has the advantages of fewer steps and quick separation process, is simple to operate, and can implement quick separation, thereby having wide application prospects in the aspect of serum protein separation; and the eluates are buffer solution can be easily prepared. In Formula (I), n is a positive integer.

Description

(1) Technical field [0001] The invention relates to a composite crystal gel continuous bed medium and a preparation method thereof, and the application of the composite crystal gel continuous bed medium to rapidly separate immunoglobulin IgG and serum albumin from human serum. (2) Background technology [0002] Human serum antibodies and albumin have important applications in many fields such as biotechnology, clinical medicine, and diagnosis. Serum is currently the main source of IgG and albumin. Existing separation methods such as salting out, ethanol precipitation, metal ion affinity chromatography, dye affinity chromatography, Protein A affinity chromatography, amino acid ligand affinity chromatography and other methods have been carried out in the separation of IgG and albumin. Some of the technologies have been applied. However, these methods often encounter problems such as easy leakage of the affinity ligand, binding of the ligand to the target protein and entering ...

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Problems solved by technology

Literature (Bereli N., et al., Materials Science and Engineering 2010, 30: 323–329) used triazine dyes Cibacron Blue F3GA and Cu 2+ Affinity crystal gel chromatography separates IgG and albumin, but these affinity media have low selectivity for these two target proteins, and the mixture of these two proteins is obtained, and it is difficult to separate the two into independent products. The adsorption capacity is low

Literature (Alkan et al., Biochemical Engineering Journal 2009, 45:201–208) used Protein A affinity crystal gel medium chromatography to separate IgG, but because Protein A is very expensive, and there is a problem of ligand leakage and contamination of the target protein, Difficult to achieve large-scale separation

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