A cell membrane chromatography column based on immobilized snap-tag fusion protein and its preparation method

Abstract

The invention discloses a cell membrane chromatographic column based on immobilized SNAP-tag fusion protein and a preparation method thereof, and belongs to the technical field of preparation of cellmembrane chromatographic columns. The method is to firstly bond benzyl guanine to silica gel, use high specific recognition and stable covalent binding of SNAP-tag and benzyl guanine which is a substrate of the SNAP-tag to enable a cell membrane to immobilize a benzyl guanine-modified silica gel stationary phase surface to obtain a cell membrane chromatographic stationary, and finally wet-pack themixed stationary phase to form the cell membrane chromatographic column based on the immobilized SNAP-tag fusion protein. The prepared cell membrane chromatographic column based on the immobilized SNAP-tag fusion protein by the invention prolongs the column life of the cell membrane chromatographic column, improves the stability of the cell membrane chromatographic column, solves the problem of poor specificity of a conventional cell membrane chromatographic column, and lays a foundation for the commercialization of the cell membrane chromatographic column.

Description

technical field [0001] The invention belongs to the technical field of cell membrane chromatographic column preparation, and relates to a cell membrane chromatographic column based on immobilized SNAP-tag fusion protein and a preparation method thereof. Background technique [0002] Cell membrane chromatography is an effective new technology for directly screening and identifying target components from complex systems. However, as an emerging technology, it inevitably still has certain defects, which are mainly reflected in two aspects: first, the life of the cell membrane chromatography column needs to be further improved. As a kind of bio-affinity chromatography, the cell membrane has all biological The general characteristics of affinity chromatography, that is, it is difficult for biological materials to maintain activity, and because the cell membrane and silica gel are mainly combined through hydrophobic interactions, the column efficiency decreases rapidly and the lif...

AI Technical Summary

Problems solved by technology

However, as an emerging technology, it inevitably still has certain defects, which are mainly reflected in two aspects: first, the life of the cell membrane chromatography column needs to be further improved. As a kind of bio-affinity chromatography, the cell membrane has all biological The general characteristics of affinity chromatography, that is, it is difficult for biological materials to maintain activity, and because the cell membrane and silica gel are mainly combined through hydrophobic interactions, the column efficiency decreases rapidly and the life is often short. When used, the biological material on it, that is, the cell membrane, will gradually fall off and inactivate, resulting in its effective service life of less than 72 hours, which greatly hinders the commercial application of cell membrane chromatography columns.

The second is that the specificity of cell membrane chromatography is poor. As a kind of bio-affinity chromatography, the activity screening function of cell membrane chromatography is realized by the bioactive material carried on it, that is, the cell membrane.

[0004] However, there are few reports on cell membrane chromatography columns based on immobilized SNAP-tag fusion proteins and their preparation methods.

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