Nickel magnetic microsphere and preparation method and application thereof

Abstract

The invention relates to a nickel magnetic microsphere and a preparation method and an application thereof. A magnetic silicon dioxide microsphere is prepared by adopting a sol-gel method, wherein themagnetic silicon dioxide microsphere takes magnetic substrate particles as a core, takes tetraethoxysilane as a silicon source and forms a shell through a reaction; the microsphere, used as a template, is modified by epoxy silane coupling agent KH-560 and iminodiacetic acid to form a ligand which is used as a complexing agent for chelating metal nickel ions to prepare the magnetic silicon dioxidemicrosphere with chelated nickel ions on the surface. By adopting the iminodiacetic acid modified silicon dioxide microsphere, multiple coordination sites can be provided for metal nickel ions, so that relatively high acting force between the modified microspheres and the metal nickel ion can be ensured, and the unsaturated coordination sites of metal ions can be acted with protein, thereby realizing that the magnetic silicon dioxide microsphere with chelated nickel ions on the surface can separate protein from a solution.

Description

technical field [0001] The invention relates to a magnetic microsphere, in particular to a nickelized magnetic microsphere, a preparation method and an application thereof. Background technique [0002] Currently, the commercially available porous agarose microspheres with a certain degree of cross-linking are mainly used for the purification of His-tagged proteins, which have a high surface area, allow proteins to contact each other, and can accommodate a large amount of liquid. However, when purifying a large amount of protein, this porous structure will also bring some problems, such as: 1) Antibodies are trapped in the pores and are difficult to wash off, so a lot of washing is required to reduce the background; 2) Washing of immunoprecipitation It is carried out in microcentrifuge tubes, and the exchange between liquids is done through a pipette gun, which is easy to lose samples. 3) The contact between the antibody on the microsphere and the target protein is slow, re...

AI Technical Summary

Problems solved by technology

However, when purifying a large amount of protein, this porous structure will also bring some problems, such as: 1) Antibodies are trapped in the pores and are difficult to wash off, so a lot of washing is required to reduce the background; 2) Washing of immunoprecipitation It is carried out in a microcentrifuge tube, and the exchange between liquids is done by a pipette gun, which is easy to lose samples

3) The contact between the antibody on the microsphere and the target protein is slow, requiring a long incubation time

4) Long incubation time and extensive washing lead to mechanical and biological (protease hydrolysis) loss of the target protein

However, it mainly loads a certain amount of magnetic substances on the basis of traditional agarose microspheres, which is cumbersome to prepare, and the structure and performance of the microspheres are not uniform.

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