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

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 ...

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 spheres of metal ion-immobilized affinity materials. And material properties are unstable, operation is unsafe, operation process is cumbersome, and other auxiliary instruments such as liquid phase pumps or centrifuges are often needed when the prepared materials are used, and the use is inconvenient and quick, etc.

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