Mutant protein A (Protein A) affinity chromatography medium

Abstract

The invention discloses a mutant protein A (Protein A) affinity chromatography medium, and the mutant protein A (Protein A) affinity chromatography medium as a chromatographic medium is used for separation and purification of antibodies and Fc fusion proteins. The mutant protein A (Protein A) affinity chromatography medium is prepared by modifying with a hydrophilic dendritic macromolecule, activating, coupling with a protein A (Protein A) ligand and capping based on genetically-engineered-modified Staphylococcus A protein, agarose or dextran gel microspheres. The modified protein A chromatographic medium has stable structure, moderate hardness, and a particle size distribution within a certain range. Therefore, modified protein A chromatographic medium has has good repeatability, high mechanical strength, greatly-improved flow rate, low cost, and large size when used in column loading, and is suitable for large-scale wide industrialization use.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a mutein A (Protein A) affinity chromatography medium. Background technique [0002] In recent years, antibody drugs have developed rapidly. In 2018, 6 of the top 10 drugs in global sales were antibody drugs, and the top 5 were all antibody drugs. The purification of antibody drugs usually adopts the classic three-step or two-step chromatography method, while the protein A (Protein A) affinity medium uses the unique and specific adsorption ability of multiple regions on the protein A (Protein A) ligand for the Fc segment of the antibody , can be used as an antibody capture step, a one-step process can remove most of the impurities in the cell fermentation broth, and the purity can reach more than 99%. With the development of antibody drugs, the affinity chromatography media used for antibody capture has also achieved rapid development. [0003] However, protein A (Protein ...

AI Technical Summary

Problems solved by technology

For example, GE's MabSelect SuRe, whose matrix is ​​highly cross-linked agarose microspheres, has the advantages of excellent hydrophilicity and good biocompatibility, which is very suitable for the separation and purification of antibody biomacromolecules; but the disadvantages are: first, The structure is soft, the particle size distribution is wide, and there are problems such as poor packing repeatability and low mechanical strength in the actual use process; second, Mabselect Sure is generally cleaned with 0.1M NaOH in biopharmaceuticals, and its alkali resistance performance still has a gap ; Thirdly, the dynamic loading capacity of MabSelect Sure is >30g / L, and there is still a little gap in the demand for large-scale antibody production

However, a fatal flaw in the structure of glass beads is that it cannot withstand the gold standard of high-concentration NaOH cleaning, which limits its application in large-scale biological samples.

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