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Metal chelating magnetic microspheres and preparation method thereof

A technology of magnetic microbeads and metal chelation, which is applied in the preparation methods of peptides, alkali metal compounds, chemical instruments and methods, etc., can solve the problems of high cost and low protein extraction efficiency, and achieves improved resistance and protection of magnetic response. The effect of high performance and high extraction efficiency

Active Publication Date: 2019-09-10
WUHAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In view of this, the present invention aims to design and prepare a novel metal chelating magnetic microbead to solve the problems of low protein extraction efficiency and high cost of existing metal chelating magnetic beads

Method used

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  • Metal chelating magnetic microspheres and preparation method thereof
  • Metal chelating magnetic microspheres and preparation method thereof
  • Metal chelating magnetic microspheres and preparation method thereof

Examples

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Embodiment 1

[0033] A metal chelating magnetic microbead, comprising a nanometer ferroferric oxide magnetic core; the surface of the nanometer ferroferric oxide magnetic core is coated with silicon dioxide; the surface of the silicon dioxide is modified with ligands for chelating metal ions; The body is composed of silane coupling agent, epichlorohydrin and iminodiacetic acid.

[0034] Among them, the silane coupling agent is γ-aminopropyltriethoxysilane (KH550).

[0035] The preparation method of above-mentioned metal chelate magnetic microbead specifically comprises the following steps:

[0036] 1) Preparation of nano-ferroferric oxide magnetic core: 1.90g of Fe(NH 4 ) 2 ·(SO 4 ) 2 ·6H 2 O, 2.33g of FeCl 3 ·6H 2 O to the three-neck flask, add 50mL deoxygenated deionized water to fully dissolve, under the protection of nitrogen, heat in a water bath at a heating temperature of 60°C for 30min, stir at a stirring rate of 200-300r / min during the heating process in the water bath and a...

Embodiment 2

[0041] A metal chelating magnetic microbead, comprising a nanometer ferroferric oxide magnetic core; the surface of the nanometer ferroferric oxide magnetic core is coated with silicon dioxide; the surface of the silicon dioxide is modified with ligands for chelating metal ions; The body is composed of silane coupling agent, epichlorohydrin and iminodiacetic acid.

[0042] Among them, the silane coupling agent is γ-aminopropyltriethoxysilane (KH550).

[0043] The preparation method of above-mentioned metal chelate magnetic microbead specifically comprises the following steps:

[0044] 1) Preparation of nano-ferroferric oxide magnetic core: 1.90g of Fe(NH 4 ) 2 ·(SO 4 ) 2 ·6H 2 O, 2.33g of FeCl 3 ·6H 2 O to the three-neck flask, add 50mL deoxygenated deionized water to fully dissolve, under the protection of nitrogen, heat in a water bath at a heating temperature of 60°C for 30min, stir at a stirring rate of 200-300r / min during the heating process in the water bath and a...

Embodiment 3

[0050] The metal chelating magnetic microbead of the embodiment of the present invention 1 is used for extracting protein, and the specific extraction method when it is used for extracting protein is as follows:

[0051] 1) be the Ni of 1mol / L by the metal chelate magnetic microbead of 5mg embodiment 1 and 1ml concentration 2+ Incubate at 220rpm, 37°C for 1h to obtain chelated Ni 2+ Metal chelated magnetic microbead solution;

[0052] 2) Adsorption and chelation of Ni with a magnet 2+ The metal chelated magnetic microbeads in solution were chelated with Ni 2+ The metal chelated magnetic microbeads, suck out the excess liquid, and then wash the chelated Ni with 3ml distilled water 2+ The metal chelated magnetic microbeads are adsorbed and chelated with Ni 2+ The metal chelated magnetic microbeads were used, the excess liquid was sucked out, and the cleaning process was repeated three times to remove excess nickel ions to obtain purified chelated Ni 2+ metal chelate magneti...

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Abstract

The invention provides metal chelating magnetic microspheres and a preparation method thereof. According to the metal chelating magnetic microspheres, superparamagnetic nano ferroferric oxide is takenas a core, the surface of the superparamagnetic nano ferroferric oxide is wrapped with a layer of silicon dioxide, and then a ligand formed by a silane coupling agent, epoxy chloropropane and iminodiacetic acid is adopted to modify the surface of silicon dioxide. The silane coupling agent and the epoxy chloropropane in the ligand can be used as connecting arms to prolong the distance between metal ions and a magnetic core, nonspecific adsorption is reduced, and thus specific adsorption of the magnetic beads to label proteins is improved. Space hindrance of combination of the target protein and the magnetic beads is reduced, so that relatively high protein extraction efficiency is achieved. The iminodiacetic acid in the ligand can chelate the metal ions with strong binding force with histidine tag protein, so that the metal chelating magnetic microspheres disclosed by the invention have relatively high capability for selectively separating proteins.

Description

technical field [0001] The invention relates to the technical field of biological materials, in particular to a metal chelating magnetic microbead and a preparation method thereof. Background technique [0002] Protein is the macromolecule with the most content and the most important function in living organisms. Proteins play a vital role in understanding the internal structure and environment of organisms, industrial production of organisms, and biomedicine. Extraction, separation and purification of target proteins are the basis for protein research. There are many methods for extracting protein by using the size, solubility and charge of protein molecules. For example, methods such as density gradient centrifugation, ultrafiltration, and gel filtration can be used to utilize the size of protein molecules, methods such as isoelectric point precipitation can be used to utilize different solubility, and polyacrylamide gel electrophoresis and ion exchange chromatography ca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/22B01J20/28B01J20/30C07K1/14
CPCB01J20/22B01J20/28009B01J20/28019C07K1/145
Inventor 谢浩范瑞文贾睿靓刘泽文曾凯孙恩杰邢佳彬李呈祥
Owner WUHAN UNIV OF TECH
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