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A kind of hydrophilic chitosan monolithic material and its preparation and application

A monolithic material, chitosan technology, applied in the field of glycoproteomics, can solve the problem of difficulty in analyzing glycoproteins, and achieve the effects of low cost, good specific adsorption effect, and strong application potential.

Active Publication Date: 2021-10-15
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the current proteomics technology has made it easy to identify proteins on a large scale, it is still difficult to directly analyze glycoproteins from complex samples

Method used

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  • A kind of hydrophilic chitosan monolithic material and its preparation and application
  • A kind of hydrophilic chitosan monolithic material and its preparation and application
  • A kind of hydrophilic chitosan monolithic material and its preparation and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Dissolve 100mg chitosan in 10mL 1% acetic acid solution, then add 5mg polyethylene glycol diglycidyl ether (Mn=500, CAS: 72207-80-8 ), the mixed solution was ultrasonicated for 1 hour, and then the mixed solution was poured into a container made of aluminum foil, slowly contacted with liquid nitrogen until completely frozen, and the resulting solid was placed in a vacuum freeze dryer at -50°C for 24 hours in vacuum, Then the obtained monolithic material was placed in an oven and heated overnight at 80°C to obtain insoluble chitosan in the eluent and eluent (acetonitrile aqueous solution containing 1% TFA) used in the enrichment of glycopeptides by hydrophilic chromatography Hydrophilic macroporous monolithic material, the material formed by SEM scanning electron microscope observation is a honeycomb material with a pore diameter of about 10um.

[0025] The prepared monolith was analyzed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MAL...

Embodiment 2

[0029] A total of 270 glycosylation sites and 400 N-glycopeptides from 146 glycoproteins were identified from 2uL human serum hydrolyzate by using the prepared monolithic material to be analyzed by a combined quadrupole Orbitrap mass spectrometer (QExactive). . The effect exceeds the 194 N-glycopeptides from 104 glycoproteins identified in the 2uL human serum enzymatic solution reported in the literature (document 6.Heet.al "High Strength and Hydrophilic Chitosan Microspheres for the Selective Enrichment of N-Glycopeptides" ANALYTICAL CHEMISTRY, 2017, 89(18):9712-9721.).

[0030] Figure 7 Venn diagram of glycosylation sites identified from 2uL IgG human serum hydrolyzate of chitosan hydrophilic macroporous monolith.

[0031] Figure 8 Venn diagram of glycoproteins identified from 2uL IgG human serum hydrolyzate of chitosan hydrophilic macroporous monolith.

[0032] Figure 9 Venn diagram of N-glycopeptide identified from 2uL IgG human serum hydrolyzate of chitosan hydrop...

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Abstract

The invention relates to preparation of chitosan hydrophilic macroporous monolithic material by freeze-drying method and application thereof. The specific preparation process uses chitosan as a monomer, polyethylene glycol diglycidyl ether as a crosslinking agent, and acetic acid solution as a solvent to prepare the chitosan hydrophilic monolithic material by freeze-drying. The overall material of the present invention is low in cost, green and environmentally friendly, has good specific adsorption effect on glycopeptides in complex samples, and has strong application potential in glycoproteomics research.

Description

technical field [0001] The invention belongs to the field of glycoproteomics, and specifically relates to the preparation and application of a chitosan hydrophilic monolithic material prepared by a freeze-drying method. The hydrophilic monolithic material can be used for the hydrophilicity of standard proteins and glycopeptides in complex biological samples. Chromatographic enrichment. Background technique [0002] Protein glycosylation is one of the most important post-translational modifications. Protein glycosylation refers to the process in which proteins are linked to sugar chains under the action of enzymes, and plays a key regulatory role in many important biological processes (ref. 1. Helenius et.al "Intracellular functions of N-linked glycans". Science, 2001, 291:2364-2369.). Glycosylation of proteins greatly increases the diversity and complexity of proteins, and endows glycoproteins with more biological functions and more important biological significance. Abnor...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/24B01J20/30
CPCB01J20/24
Inventor 欧俊杰张路伟马淑娟叶明亮于之渊姜利孙传盛
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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