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Peptide identification method by using mesoporous silica composite combined with mass spectrum

A technology of mesoporous silica and composite materials, applied in chemical instruments and methods, alkali metal compounds, alkali metal oxides/hydroxides, etc., to achieve the effects of large specific surface area, strong affinity, and high selectivity

Inactive Publication Date: 2016-03-02
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional affinity-based enrichment strategies are only suitable for a certain type of protein / peptide with specific functional groups, such as metal oxide affinity chromatography for the enrichment of phosphorylated peptides, and boronic acid affinity chromatography for sugars Peptide enrichment, etc.

Method used

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  • Peptide identification method by using mesoporous silica composite combined with mass spectrum
  • Peptide identification method by using mesoporous silica composite combined with mass spectrum
  • Peptide identification method by using mesoporous silica composite combined with mass spectrum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Synthesis of a perfluoroalkyl-modified mesoporous silica composite material coated on the surface of a magnetic microsphere ferric oxide

[0033] (1) 1.35g ferric chloride hexahydrate (FeCl 3 ·6H 2 O) Dissolve in 75mL of ethylene glycol, stir magnetically until clarified, add 3.6g of sodium acetate, fully stir until dissolved, continue stirring for 0.5h, transfer to a hydrothermal synthesis reaction kettle, ultrasonicate for 5 minutes, then put in an oven Heat at 200°C for 16 hours, take out the reaction kettle, and cool for 10 hours; pour out the magnetic balls obtained from the reaction from the reaction kettle, wash them thoroughly with deionized water and absolute ethanol, and dry them under vacuum at 50°C;

[0034] (2) Disperse 75 mg of magnetic balls obtained in step (1) and 750 mg of cetyltrimethylammonium bromide in 75 mL of deionized water, and ultrasonicate for 30 minutes; -3 M sodium hydroxide solution was mixed, and a uniform and stable dispersi...

Embodiment 2

[0040] Example 2: The surface of the magnetic microsphere ferric oxide obtained in Example 1 is coated with a perfluoroalkyl-modified mesoporous silica composite material as a solid-phase microextraction adsorbent for the low concentration of perfluoroalkyl derivatization Enrichment and MALDI-TOFMS detection of β-casein hydrolyzate.

[0041] (1) Preparation of standard protein enzymatic hydrolysis solution: Accurately weigh 5mg of standard protein β-casein and dissolve in 25mM ammonium bicarbonate buffer, boil for 10 minutes, dilute to 1mg / mL with 25mM ammonium bicarbonate buffer, and then mix with protein Add an appropriate amount of trypsin at a volume ratio of 1:40, and enzymatically hydrolyze at 37°C for 16 hours.

[0042] (2) Derivation of standard proteolysis solution: Add 10 μL of dimethyl sulfoxide / ethanol solution (volume ratio 3:1) and 9.2 μL of saturated barium hydroxide solution to 10 μL of β-casein solution obtained in step (1) , 2 μL of sodium hydroxide solution...

Embodiment 3

[0048] Example 3: Synthesis of a perfluoroalkyl-modified mesoporous silica composite material coated on the surface of a magnetic microsphere ferric oxide

[0049] (1) 1.35g ferric chloride hexahydrate (FeCl 3 ·6H 2 O) Dissolve in 70mL of ethylene glycol, stir magnetically until clarified, add 5.0g of sodium acetate, stir fully until dissolved, continue stirring for 0.5h, transfer to a hydrothermal synthesis reaction kettle, ultrasonicate for 5 minutes, then put in an oven Heat at 190°C for 18 hours, take out the reaction kettle, and cool for 12 hours; pour out the magnetic balls obtained from the reaction from the reaction kettle, wash them thoroughly with deionized water and absolute ethanol, and dry them under vacuum at 60°C;

[0050] (2) Disperse 50 mg of magnetic balls obtained in step (1) and 500 mg of cetyltrimethylammonium bromide in 50 mL of deionized water, and ultrasonicate for 30 minutes; -3 M sodium hydroxide solution was mixed, and a uniform and stable dispersi...

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Abstract

The invention provides a peptide identification method by using a mesoporous silica composite combined with mass spectrum. A composite with perfluoroalkyl group modified mesoporous silica covering on the surface of Fe3O4 magnetic microsphere is used in matrix-assisted laser desorption ionization-time of flight mass spectrometric analysis of perfluoroalkyl group derived peptide. The material shows extremely strong affinity with the fluorous phase derived phosphorylated peptides, while maintaining good dispersion in aqueous solution. The Fe3O4 magnetic core in the material simplifies the process of enrichment and separation. A solid phase microextraction method based on the composite with perfluoroalkyl group modified mesoporous silica covering on the surface of Fe3O4 magnetic microsphere achieves selective enrichment of non-phosphorylated peptides, and can be successfully applied to the enrichment of endogenous phosphopeptides in human serum.

Description

technical field [0001] The invention belongs to the field of advanced nanomaterials and biotechnology, and relates to a method for identifying peptide segments by combining mesoporous silica composite material with mass spectrometry, and in particular to a magnetic microsphere ferric oxide surface coated with polyperfluoroalkyl modification Application of Mesoporous Silica Composite Materials for Enrichment and MALDI-TOFMS Analysis of Perfluoroalkyl-Derivatized Phosphorylated Peptides. Background technique [0002] Perfluoroalkyl groups can produce delocalization effects in fluorine-containing environments. Fluorine-fluorine solid-phase microextraction takes advantage of this feature. Through perfluoroalkyl-modified media, substances containing perfluoroalkyl groups are separated from fluorine-free substances. matter separated. Fluorine-fluorine solid-phase microextraction technology is usually used in targeted synthesis, heterogeneous catalysis and separation and purificat...

Claims

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Application Information

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IPC IPC(8): B01J20/26B01J20/28B01J20/281B01J20/30
Inventor 邓春晖赵曼
Owner FUDAN UNIV
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