Hydrophilic metal ion immobilization affinity magnetic bead and preparation and application thereof

A metal ion and hydrophilic technology, which is applied in the direction of microsphere preparation, peptide preparation methods, chemical instruments and methods, etc., can solve the problem of poor hydrophilicity of metal ion-immobilized affinity materials, unstable performance of affinity materials, preparation Problems such as poor process reproducibility, to achieve the effect of less steps, good hydrophilicity and good reproducibility

Active Publication Date: 2012-10-31
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a hydrophilic metal ion-immobilized affinity magnetic ball and its preparation and application, in order to solve the problem of poor hydrophilicity, poor selectivity, poor reproducibility of the preparation process, and the obtained affinity magnetic sphe

Method used

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  • Hydrophilic metal ion immobilization affinity magnetic bead and preparation and application thereof
  • Hydrophilic metal ion immobilization affinity magnetic bead and preparation and application thereof
  • Hydrophilic metal ion immobilization affinity magnetic bead and preparation and application thereof

Examples

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Effect test

Embodiment 1

[0022] Example 1: Preparation of a hydrophilic titanium ion-immobilized affinity nanomagnetic sphere

[0023] 0.1 g of amino magnetic spheres (particle size of 200 nm) and 5 ml of glutaraldehyde solution with a mass concentration of 20% were reacted at room temperature for 2 h, and the pH of the solution was 5 to activate the surface of the amino magnetic spheres. After the activation reaction on the surface of the magnetic ball is completed, wash the magnetic ball with a solution with a pH value of 5 for 3 times. Add the activated amino magnetic spheres to 3 ml of adenosine triphosphate disodium solution with a mass concentration of 200 mg / mL, the pH of the solution is 5, react at room temperature for 2 h, and carry out the chemical modification reaction on the surface of the magnetic spheres to make the magnetic spheres Hydrophilic functional molecules containing multiple phosphate groups and hydroxyl groups are bonded to the surface for subsequent fixation of titanium ions....

Embodiment 2

[0025] Example 2: Preparation of a hydrophilic gallium ion-immobilized affinity nanomagnetic sphere

[0026] The metal ion solution is a gallium nitrate solution, and the others are the same as in Example 1 to obtain hydrophilic gallium ion-immobilized affinity nanomagnetic balls.

Embodiment 3

[0027] Example 3: Preparation of a hydrophilic iron ion-immobilized affinity nanomagnetic sphere

[0028] The metal ion solution is a ferric chloride solution, and the others are the same as in Example 1 to obtain hydrophilic iron ion-immobilized affinity nano-magnetic balls.

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Abstract

The invention relates to a preparation method of a hydrophilic metal ion immobilization affinity magnetic bead, which comprises the following steps: the surface of an amino magnetic bead is subjected to amino activation through glutaral; the activated magnetic bead is connected with an adenosine disodium triphosphate molecule through a surface modification reaction; the surface of the activated magnetic bead is bonded with a hydrophilic functional molecule containing a plurality of phosphate and hydroxyl radicals, so as to be beneficial for subsequent metal ion immobilization; at last, the immobilization of a metal ion is realized in a solution containing the metal ion; and the preparation of the hydrophilic metal ion immobilization affinity magnetic bead is accomplished. The hydrophilic metal ion immobilization affinity magnetic bead has the advantages that the preparation method is simple; compared with other metal ion immobilization affinity chromatographic columns or magnetic beads, the hydrophilic metal ion immobilization affinity magnetic bead prepared by the method is high in hydrophily, good in selectivity, good in preparation repeatability and stable in performance, and has a good specific concentration ability to phosphorylated peptides; the operation is simple; the complicated centrifugation operation can be avoided; and the hydrophilic metal ion immobilization affinity magnetic bead has wide application prospects in proteomics, especially in phosphorylated proteomics.

Description

technical field [0001] The invention relates to a hydrophilic magnetic sphere for selective enrichment and separation of phosphorylated peptides on-line or off-line, in particular to a hydrophilic metal ion-immobilized affinity magnetic sphere and its preparation and application. Background technique [0002] Protein phosphorylation is one of the most common post-translational modifications, which plays an important role in signal transduction, metabolic regulation, and cell function regulation in life processes. However, phosphorylated proteins / peptides are usually present in low amounts and are often interfered by large amounts of non-phosphorylated proteins / peptides in mass spectrometry analysis. Therefore, the development of selective enrichment technology for phosphorylated peptides is very important for in-depth research on phosphorylated proteomes. [0003] Immobilized metal affinity chromatography based on the specific interaction between immobilized metal ions and ...

Claims

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Application Information

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IPC IPC(8): H01F1/00B01J13/02C07K1/22B82Y25/00B82Y40/00
Inventor 张丽华张丽媛杨开广梁振张玉奎
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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