Method for preparing iron ion immobilization affinity chromatography monolith column

Abstract

The invention relates to a method for preparing an iron ion immobilization affinity chromatography monolith column, which comprises: preparing porous monolith column substrate by polymerization in capillaries with N-hydroxysuccinimide serving as a functional monomer, ethanedimethylethyl acrylate as a crosslinking agent, mixture of cyclohexanol, dodecanol and N,N-dimethylformamide as a porogen andazodiisobutyronitrile as an initiator; bonding carboxyl-containing functional groups on the surface of the monolith column substrate through surface modification for subsequent iron ion immobilization; and immobilizing iron ions by using iron ion containing solution to accomplish the preparation of the iron ion immobilization affinity chromatography monolith column. The invention has the advantages that: the preparation method is simple and convenient; and the micro structure of the obtained substrate material is uniform. Compared with other metal ion immobilization affinity chromatography monolith column, the iron ion immobilization affinity chromatography monolith column prepared by the method has high preparation repeatability, and has stable performance and high specific enriching capacity for phosphorylated peptide fragments.

Description

Technical field [0001] The invention relates to an immobilized affinity chromatographic monolithic column for the selective enrichment and separation of online or offline phosphorylated peptides, in particular to the preparation of an iron ion immobilized affinity chromatographic monolithic column. Background technique [0002] Protein phosphorylation is one of the most common post-translational modifications, and it plays an important role in signal transduction, metabolic regulation, and cell function regulation during life. However, phosphorylated proteins / peptides are usually low in content and are often interfered by a large number of non-phosphorylated proteins / peptides in mass spectrometry analysis. Therefore, the development of selective enrichment techniques for phosphorylated peptides is essential for in-depth research on phosphorylated proteome. [0003] Immobilized metal affinity chromatography based on the specific interaction between immobilized metal ions and phosph...

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a method for preparing an iron ion-immobilized affinity chromatography monolithic column, in order to solve the problem of long chemical modification time, cumbersome operation process and the problem of immobilized metal ion affini

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