A kind of metal chelating magnetic microbead and preparation method thereof

A technology of magnetic microbeads and metal chelation, which is applied to the preparation method of peptides, alkali metal compounds, chemical instruments and methods, etc., can solve the problems of low protein extraction efficiency and high cost, and achieve the protection of magnetic responsiveness and improvement of resistance , reduce the effect of non-specific adsorption

Active Publication Date: 2022-08-05
WUHAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • A kind of metal chelating magnetic microbead and preparation method thereof
  • A kind of metal chelating magnetic microbead and preparation method thereof
  • A kind of metal chelating magnetic microbead and preparation method thereof

Examples

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Embodiment 1

[0033] A metal chelating magnetic microbead, comprising a nanometer triferromagnetic core; the surface of the nanometer triferromagnetic core is wrapped with silicon dioxide; the surface of the silicon dioxide is modified with a ligand for chelating metal ions; The body consists of silane coupling agent, epichlorohydrin and iminodiacetic acid.

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

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

[0036] 1) Preparation of nanometer ferric oxide magnetic core: 1.90g Fe(NH 4 ) 2 ·(SO 4 ) 2 ·6H 2 O, 2.33g FeCl 3 ·6H 2 0 to a three-necked flask, add 50 mL of deoxygenated deionized water to fully dissolve, under nitrogen protection, heat in a water bath at a heating temperature of 60 ° C for 30 min, stir at a stirring rate of 200-300 r / min during the heating process in the water bath and adjust the pH with 45...

Embodiment 2

[0041] A metal chelating magnetic microbead, comprising a nanometer triferromagnetic core; the surface of the nanometer triferromagnetic core is wrapped with silicon dioxide; the surface of the silicon dioxide is modified with a ligand for chelating metal ions; The body consists of silane coupling agent, epichlorohydrin and iminodiacetic acid.

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

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

[0044] 1) Preparation of nanometer ferric oxide magnetic core: 1.90g Fe(NH 4 ) 2 ·(SO 4 ) 2 ·6H 2 O, 2.33g FeCl 3 ·6H 2 0 to a three-necked flask, add 50 mL of deoxygenated deionized water to fully dissolve, under nitrogen protection, heat in a water bath at a heating temperature of 60 ° C for 30 min, stir at a stirring rate of 200-300 r / min during the heating process in the water bath and adjust the pH with 45...

Embodiment 3

[0050] The metal chelated magnetic microbeads of Example 1 of the present invention are used for protein extraction, and the specific extraction method when it is used for protein extraction is as follows:

[0051] 1) Mix 5 mg of the metal chelated magnetic microbeads of Example 1 with 1 ml of Ni with a concentration of 1 mol / L. 2+ Incubate for 1 h at 220 rpm and 37 °C to obtain chelated Ni 2+ The metal chelated magnetic microbead solution;

[0052] 2) Use a magnet to adsorb chelated Ni 2+ The metal-chelated magnetic microbead solution chelates Ni 2+ The metal chelated magnetic microbeads, aspirated the excess liquid, and then washed the chelated Ni with 3 ml of distilled water. 2+ metal-chelated magnetic microbeads, and adsorbed with magnets chelated with Ni 2+ The metal chelated magnetic microbeads were obtained, the excess liquid was sucked out, and the cleaning process was repeated three times to remove excess nickel ions, and purified chelated Ni ions were obtained. ...

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Abstract

The invention provides a metal chelated magnetic microbead and a preparation method thereof. The metal chelated magnetic microbead takes a superparamagnetic nanometer ferric oxide as a core, and wraps a layer of silicon dioxide on its surface. The surface of silica is modified by the ligand formed by silane coupling agent, epichlorohydrin and iminodiacetic acid. The silane coupling agent and epichlorohydrin in the ligand can be used as connecting arms to extend the connection between metal ions and magnetic cores. distance, reducing non-specific adsorption, thereby improving the specific adsorption of labeled proteins by magnetic beads, and reducing the steric hindrance of target protein binding to magnetic beads, making it have higher protein extraction efficiency, while iminodiacetic acid in the ligand can chelate The metal ions with strong binding force to histidine-tagged proteins make the metal chelated magnetic microbeads of the present invention have a higher ability to selectively separate proteins.

Description

technical field [0001] The invention relates to the technical field of biological materials, in particular to a metal chelated magnetic microbead and a preparation method thereof. Background technique [0002] Proteins are the most abundant and functional macromolecules in organisms. Proteins play a vital role in understanding the internal structure and environment of organisms, bioindustrial production and biomedicine. Extraction, separation and purification of target proteins are the basis of protein research. There are many methods for extracting proteins by using the different sizes, solubility and charge of protein molecules. For example, density gradient centrifugation, ultrafiltration, gel filtration and other methods can be used to use the size of protein molecules, isoelectric precipitation and other methods can be used to use different solubility, and polyacrylamide gel electrophoresis and ion exchange chromatography can be used to use different electric charges....

Claims

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Application Information

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Patent Type & Authority Patents(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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