Preparation method of metal-chelating magnetic beads capable of removing metal ions from macromolecules

A metal chelation and metal ion technology, applied in the directions of alkali metal compounds, chemical instruments and methods, alkali metal oxides/hydroxides, etc., can solve problems such as inability to remove metal ions, and achieve easy amplification and simple preparation process. , easy to use effect

Inactive Publication Date: 2015-12-02
HENAN UNIV OF SCI & TECH
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Problems solved by technology

Chinese patent application "a metal chelated nano-magnetic bead, its preparation method and its application" (application number CN200610002030.1) discloses a nano-magnetic bead connected with IDA. After the magnetic bead is chelated with metal ions, the metal ions (such as six-coordinated metals Ni, Cu, Co, and Ca, etc.) there are three empty orbitals that can bind to proteins with metal binding sites, so nano-magnetic beads made of IDA as a chelating group have a strong ability to adsorb proteins , can be used well for the separation and identification of metal-binding proteins in solution, but cannot effectively remove metal ions from macromolecules

Method used

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preparation example Construction

[0015] A method for preparing metal chelate magnetic beads capable of removing metal ions from macromolecules, comprising the following steps:

[0016] Step 1. Activation of hydroxyl magnetic beads: adding lye to the hydroxy magnetic beads for activation to obtain a mixture I containing activated hydroxy magnetic beads and lye for later use; the amount of lye added is such that the pH of the mixture I Value reaches 12-14;

[0017] Step 2, epoxidation: according to the volume ratio of the mixture Ⅰ, dimethyl sulfoxide, epichlorohydrin and sodium hydroxide solution is 1:2:1:4, add dimethyl to the mixture Ⅰ obtained in the above step 1 Mix sulfoxide, epichlorohydrin and sodium hydroxide solution, mix and stir slowly for 12-14 hours at a temperature of 60°C, make the activated hydroxyl magnetic beads connect to epoxy groups, remove the solution, and wash the obtained magnetic beads to medium property, that is, to obtain epoxy activated magnetic beads, for subsequent use;

[0018...

Embodiment 1

[0024] Add 6mol / L sodium hydroxide solution to 1ml of 10mg / ml hydroxyl magnetic beads and fully activate to obtain a mixture I containing activated hydroxyl magnetic beads and lye; the amount of 6mol / L sodium hydroxide solution is to make the pH of mixture The value reaches 12; add 2ml dimethyl sulfoxide, 1ml epichlorohydrin and 4ml 1mol / L sodium hydroxide solution to the mixture plus I (1ml epichlorohydrin and 4ml sodium hydroxide are slowly stirred and divided into 5 parts within 14 hours. Adding each time with an interval of 2.8 hours), stirring slowly at 60°C overnight and then removing the solution, the obtained magnetic beads were first washed with water, then washed with dilute acetic acid, and finally washed with water to obtain epoxy-activated magnetic beads. Then add 4ml concentration of 1mol / L sodium carbonate solution, 0.1g of EDDS-Na to the epoxy activated magnetic beads 3 and 1 mg NaBH 4 , stirred slowly at 50°C for 14 hours, removed the solution, and obtained h...

Embodiment 2

[0026] After adding 5mol / L sodium hydroxide solution to 1ml10mg / ml hydroxyl magnetic beads and fully activating, the mixture I containing activated hydroxyl magnetic beads and lye is obtained; the amount of 5mol / L sodium hydroxide solution is to make the pH of mixture I The value reaches 14; Add 2ml dimethyl sulfoxide, 1ml epichlorohydrin and 4ml 1mol / L sodium hydroxide solution to the mixture plus I (1ml epichlorohydrin and 4ml sodium hydroxide solution are slowly stirred and divided within 13 hours. Add 5 times, each interval 2.6 hours), slowly stir overnight at 60°C, then wash with water, dilute acetic acid, and water to obtain epoxy-activated magnetic beads. Then add 4ml concentration of 5mol / L sodium carbonate solution, 0.5gEDDS-Na 3 , 3mgNaBH 4 , stirred slowly at 40 °C for 15 hours. Wash the magnetic beads connected with EDDS with water first, then wash with dilute acetic acid, and finally wash with water until neutral to obtain metal chelate magnetic beads that can r...

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Abstract

A preparation method of metal-chelating magnetic beads capable of removing metal ions from macromolecules comprises the steps that alkali liquor is added into the hydroxyl magnetic beads to perform activation so as to obtain a mixture I containing the activated hydroxyl magnetic beads and the alkali liquor; dimethyl sulfoxide, epoxy chloropropane and a sodium hydroxide solution are added into the mixture I, slow stirring is performed at the temperature of 60 DEG C for 12-14 hours after mixing to obtain epoxy activated magnetic beads; a sodium carbonate solution, N,N'-1,2-ethanediylbis-1-aspartic acid and sodium borohydride are added into the epoxy activated magnetic beads, slow stirring is performed at the temperature of 40-60 DEG C for 10-15 hours after mixing to obtain the magnetic beads capable of removing the metal ions from the macromolecules. The preparation method is simple in preparation process and facilitates enlargement, heavy metal ions in solutions and biological macromolecules can be quickly removed after the magnetic beads are added into the solutions, and analysis on structures and functions of the biological macromolecules is facilitated.

Description

technical field [0001] The invention relates to a metal chelating magnetic bead, in particular to a method for preparing a metal chelating magnetic bead capable of removing metal ions from macromolecules. Background technique [0002] In solution, metal ions are often combined with biomacromolecules such as proteins, nucleic acids, sugars, etc., affecting our analysis of the structure and function of these biomacromolecules. Existing methods for demetallization are by adding high concentrations of low molecular weight chelating agents, such as EDTA and EGTA, or by metal chelating agarose gel. The former removes the metal complexes of the chelating agent and at the same time the metal ions in the buffer will recombine with the macromolecules, affecting the demetallization effect. Currently, agarose gels are widely used to demetallize biomacromolecules. The particle size of agarose is uniform, and there are reactive active groups with appropriate density on the surface, whic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/22B01J20/28B01J20/30
Inventor 张红晓宋玉峰王发园侯典云
Owner HENAN UNIV OF SCI & TECH
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