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Preparation of magnetic silicon dioxide microsphere with metallic ion chelated surface and use thereof

A technology of silica and metal ions, which is applied in the field of materials for separating hexahistidine proteins, can solve problems such as metal ion omission, weak interaction force, and weak protein interaction force

Inactive Publication Date: 2008-12-17
SHAANXI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In common complexing agents, IDA is three-coordinated, and the force between it and metal ions is relatively weak, which can easily cause the problem of missing metal ions. N, N, N'-tris(carboxymethyl)ethylenediamine TED It is a five-coordinate, and has a strong interaction with metal ions. Since only one bond interacts with protein, the interaction with protein is too weak.

Method used

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  • Preparation of magnetic silicon dioxide microsphere with metallic ion chelated surface and use thereof
  • Preparation of magnetic silicon dioxide microsphere with metallic ion chelated surface and use thereof
  • Preparation of magnetic silicon dioxide microsphere with metallic ion chelated surface and use thereof

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Embodiment 1

[0036] This embodiment provides the preparation method of magnetic silica microspheres with surface chelated metal ions, and its process steps are as follows:

[0037] 1. Preparation of magnetic silica microspheres

[0038] Take 7.2892g of cetyltrimethylammonium bromide and 100.0g of toluene into a four-necked flask, the mass ratio of cetyltrimethylammonium bromide to toluene is 1:15, and use a mixer to Stir well; 0.329g FeCl 2 4H 2 O with 0.987g FeCl 3 ·6H 2 O was dissolved in 9.4397g of secondary water to prepare an aqueous solution with a concentration of 140g / L, FeCl 2 4H 2 O and FeCl 3 ·6H 2 O mass ratio is 1:3, at 2~3mL / min N 2 Add this solution dropwise to the above mixture at 4-5mL / h under air protection, and continue stirring at 800 rpm for 4-6 hours; then add 2mL of ammonia water with a mass concentration of 25%, cetyltrimethylammonium bromide The mass ratio of ammonia water with a mass concentration of 25% is 1:0.17, and the stirring is continued at 800 rpm...

Embodiment 2

[0047] In the preparation of magnetic silica microspheres process step 1, get 7.2892g cetyltrimethylammonium bromide and 87.47g toluene and join in the four-necked flask, cetyltrimethylammonium bromide and toluene The mass ratio is 1:12, stir evenly with a stirrer at 1000 rpm; mix 0.305g FeCl 2 4H 2 O with 0.916g FeCl 3 ·6H 2 O was dissolved in 9.4397g of secondary water to prepare an aqueous solution with a concentration of 130g / L, FeCl 2 4H 2 O and FeCl 3 ·6H 2 O mass ratio is 1:3, at 2~3mL / min N 2 Add this solution dropwise to the above mixture at 4-5mL / h under air protection, and continue stirring at 800 rpm for 4-6 hours; The mass ratio of ammonium to ammonia water with a mass concentration of 25% is 1:0.1, and the stirring is continued at 800 rpm for 2 hours. In the mixture, ferric oxide nanoparticles are formed; slowly add ethyl orthosilicate at 2 to 3 mL / h Ester 6.78mL, hexadecyltrimethylammonium bromide and tetraethyl orthosilicate in a mass ratio of 1:1, stir...

Embodiment 3

[0052] In the preparation of magnetic silica microspheres process step 1, get 7.2892g cetyltrimethylammonium bromide and 145.784g toluene and join in the four-necked flask, cetyltrimethylammonium bromide and toluene The mass ratio is 1:20, stir evenly with a stirrer at 1000 rpm; mix 0.354g FeCl 2 4H 2 O with 1.060g FeCl 3 ·6H 2 O was dissolved in 9.4397g of secondary water to prepare an aqueous solution with a concentration of 150g / L, FeCl 2 4H 2 O and FeCl 3 ·6H 2 O mass ratio is 1:3, at 2~3mL / min N 2 Add this solution dropwise to the above mixture at 4~5mL / h under air protection, and continue stirring at 800 rpm for 4~6 hours; The mass ratio of ammonium to 25% ammonia water is 1:0.2, and the mixture is continuously stirred at 800 rpm for 2 hours. In the mixture, iron ferric oxide nanoparticles are formed; slowly add ethyl orthosilicate at 2 to 3 mL / h Esters 11.76mL, hexadecyltrimethylammonium bromide and tetraethyl orthosilicate at a mass ratio of 1:1.5, stirred and ...

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Abstract

The invention relates to a method for preparing magnetic silicon dioxide microspheres with the surfaces chelated with metal ions; the method of the inventioncomprises the following steps of preparation of the magnetic silicon dioxide microspheres, epoxidation of the magnetic silicon dioxide microspheres, carboxylation of the magnetic silicon dioxide microspheres and chelation technique of metal ions. Compared with the existing preparing method, the method of the invention has the advantages of reasonable design, simple operation and large amount of chelation of metal, etc. The magnetic silicon dioxide microspheres with the surfaces chelated with the metal ions, which are prepared by adopting the method, contain cobalt ions, have stronger bonding force with the modified microspheres so as to lead the cobalt ions to be not easy to leak out, and have higher selectivity and proper bonding strength with histidine on the protein, thus leading the magnetic silicon dioxide microspheres with the surfaces chelated with the metal ions to have high selectivity to hexameric histidine tag protein, and being easy to realize the separation and the purification of the target protein; the magnetic silicon dioxide microspheres with the surfaces chelated with the metal ions, which are prepared by adopting the method, can be used for separating the hexameric histidine tag protein.

Description

technical field [0001] The invention belongs to the technical field of materials, and in particular relates to a material used for isolating hexahistidine protein from Escherichia coli. Background technique [0002] Biomagnetic separation technology is a high-efficiency separation technology developed on the basis of traditional magnetic separation technology, which is applied to organisms such as cells, bacteria, nucleic acids and proteins. It uses the characteristics of magnetic or magnetically labeled organisms to perform qualitative movements under the action of an external magnetic field to extract, enrich, separate and purify target organisms. Early work mainly focused on the development of preparation techniques for magnetic markers, including the adsorption of magnetic nanoparticles to polymer microspheres such as agarose, polyacrylamide, polyglutaraldehyde, and polyvinylaldehyde by immersion in ferrofluid. The microspheres are characterized by a uniform particle si...

Claims

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Application Information

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IPC IPC(8): C01B33/18C07K1/13
Inventor 冯国栋胡道道崔亚丽杨柳杨磊
Owner SHAANXI NORMAL UNIV
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