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Preparation of polymer material for enriching phosphorylated peptide segments

A technology of polymer materials and phosphorylated peptides, which is applied in the field of separation and purification of phosphorylated peptides, and can solve time-consuming and labor-intensive problems

Active Publication Date: 2009-04-22
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In order to prepare this chromatographic adsorption material with phosphoric acid groups, the surface of the material needs to be modified, which requires multi-step chemical reactions, so it is time-consuming and labor-intensive.

Method used

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  • Preparation of polymer material for enriching phosphorylated peptide segments
  • Preparation of polymer material for enriching phosphorylated peptide segments
  • Preparation of polymer material for enriching phosphorylated peptide segments

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Preparation of polymer monolithic column

[0026] The monolithic column is a special liquid chromatography column, which forms a porous and continuous polymer by pouring the polymer reaction liquid into the column tube through in-situ polymerization. The polymer monolithic column has very low column pressure due to many through holes, and has little mass transfer resistance due to convective mass transfer, which is very suitable for sample pretreatment. In this example, a polymer monolithic capillary column with phosphate groups was prepared and used for the enrichment of phosphopeptides.

[0027] 1) Capillary pretreatment

[0028] First wash the empty capillary column with 0.1M NaOH solution for 1 h, then rinse the capillary tube with deionized water until the pH of the effluent liquid is 7.0, then rinse the capillary column with methanol solution for 10 min, and dry it with nitrogen. A mixture of methanol and methacryloxypropyl-trimethoxysilane was injecte...

Embodiment 2

[0059] Example 2 Preparation of polymer particles

[0060] Most liquid chromatography columns and solid phase extraction columns use granular fillers such as agarose particles, organic polymer particles, and inorganic particles as stationary phases. In this example, polymer particles with phosphoric acid groups were prepared and For enrichment of phosphopeptides.

[0061] 1) Preparation of polymer particles

[0062] Put the polymerization reaction liquid (the proportion is the same as in Example 1) in a centrifuge tube, seal it and immerse it in a water bath at 60°C for 8-24 hours. After the reaction is completed, grind into particles and soak in methanol to remove porogens, residual reaction reagents and some low-polymerization substances produced by the reaction.

[0063] 2) Immobilization of zirconium ions

[0064] The prepared particles were immersed in 20mM zirconium oxychloride solution, shaken for 15 minutes, centrifuged, and the sediment was collected to obtain zirc...

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Abstract

The invention relates to high-selectivity enrichment of phosphorylated peptide, in particular to a method for preparing polymer materials for enriching phosphorylated peptide fragments. The method adopts monomers provided with phosphate groups to perform polymerization, so as to directly form the polymer materials provided with the phosphate groups. The materials are acted by a solution containing zirconium ions to generate phosphate ester zirconium groups, so that the materials can specially enrich and purify the phosphorylated peptide from a complex proteolysis solution. The method adopts the polymerization method to directly synthesize the polymer materials provided with functional groups, does not need to additionally perform chemical modification, and has the characteristics of simplicity and convenience.

Description

technical field [0001] The invention relates to the separation and purification of phosphorylated peptides, in particular to a method for preparing a polymer material for enriching phosphorylated peptides. Background technique [0002] The identification of protein post-translational modification sites is one of the important challenges in current proteomics research, and phosphorylation modification is one of the most important post-translational modifications of proteins. Because the reversible protein phosphorylation modification reaction regulates the activity and function of the protein in the aspects of cell signal transduction and metabolism. Recently, mass spectrometry has been developed as one of the important tools in the characterization of protein phosphorylation. In the mass spectrometry analysis of phosphorylated peptides of phosphorylated protein digests, due to the presence of a large number of non-phosphorylated peptides in the digested products, the ion si...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08F130/02B01J20/285
Inventor 邹汉法董靖叶明亮周厚江
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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