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Hydrophilic chitosan integral material and preparation and application thereof

A technology of monolithic material and chitosan, applied in the field of glycoproteomics, can solve problems such as difficulty in analyzing glycoproteins, and achieve the effects of low cost, good specific adsorption effect, and green cost

Active Publication Date: 2020-04-07
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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  • Hydrophilic chitosan integral material and preparation and application thereof
  • Hydrophilic chitosan integral material and preparation and application thereof
  • Hydrophilic chitosan integral material and preparation and application thereof

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 analyzing the prepared monolithic material by a combined quadrupole Orbitrap mass spectrometer (QExactive) . The effect exceeds that reported in the literature that 194 N-glycopeptides from 104 glycoproteins were identified from 2uL human serum enzymatic solution (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 hydrophil...

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Abstract

The invention relates to a hydrophilic macroporous chitosan integral material prepared by using a freeze drying method and application thereof. According to the specific preparation process of the material, chitosan is used as a monomer, polyethylene glycol diglycidyl ether is used as a cross-linking agent, an acetic acid solution is used as a solvent, and the hydrophilic chitosan integral material is prepared through the freeze-drying method. The integral material disclosed by the invention is low in cost, green and environment-friendly, has a good specific adsorption effect on glycopeptide in a complex sample, and shows good 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 Applications(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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