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Synthetic method and application of metal-organic framework composite nanomaterial

A nano-composite material and metal-organic framework technology, which is applied in the direction of alkali metal compounds, analytical materials, and material inspection products, can solve the problems of low abundance of glycopeptides and phosphorylated peptides, low ionization efficiency, and difficulties in mass spectrometry detection. Good body pore structure, improved mass spectrometry signal, high sensitivity and selectivity effects

Inactive Publication Date: 2017-03-22
FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the abundance of glycopeptides and phosphorylated peptides is often very low, and their mass spectral response will be overwhelmed by the high abundance of non-phosphorylated peptides / glycopeptides and proteins, as well as salt and surfactant in the sample. Behavior interferes, making its ionization efficiency very low, and mass spectrometry detection is relatively difficult

Method used

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  • Synthetic method and application of metal-organic framework composite nanomaterial
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  • Synthetic method and application of metal-organic framework composite nanomaterial

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Synthesis of a metal-organic framework (MOF) nanocomposite material with zirconium as the central metal ion, coated with polydopamine and amino groups on the surface of magnetic balls.

[0043] (1) Using ethylene glycol as a solvent to synthesize ferroferric oxide magnetic balls, 1.35g FeCl 3 •6H 2 O was dissolved in 75mL of ethylene glycol, stirred magnetically (sealed with gloves) until clear, then added 3.6g of crushed sodium acetate and stirred until dissolved and continued stirring for 0.5h (sealed with gloves). After ultrasonication for 5 minutes, transfer to the reaction kettle, 200°C, 16h. The reaction kettle was taken out and cooled overnight. Pour out the magnetic ball and wash it with water 5 times (sonication for 5 minutes each time). Fully wash the magnetic balls with deionized water and ethanol until the washing solution is clear and pure, and dry in vacuum at 50°C;

[0044] (2) Prepare Tris (Tris) buffer solution (solvent is deionized water...

Embodiment 2

[0057] Example 2: The surface of the magnetic ball obtained in Example 1 is coated with polydopamine and amino-modified metal-organic framework (MOF) nanocomposites with zirconium as the central metal ion as a solid-phase microextraction adsorption separation medium for low-concentration HRP The enrichment and MALDI-TOF MS detection of enzymatic hydrolyzate and β-Casein enzymolyzate.

[0058] (1) Preparation of standard protein enzymatic hydrolysis solution: Accurately weigh 2 mg of HRP standard protein, use 25 mM ammonium bicarbonate solution to prepare a standard protein solution with a concentration of 2 mg / mL, the pH is about 8.3, and boil for ten minutes. According to the ratio of trypsin to standard protein with a mass ratio of 1:50, add trypsin and incubate at 37°C for 16 hours to obtain 2 mg / mL HRP trypsin hydrolyzate; accurately weigh 4 mg IgG standard For protein, use 25 mM ammonium bicarbonate solution to make a standard protein solution with a concentration of 4 mg...

Embodiment 3

[0067] Example 3: The surface of the magnetic balls obtained in Example 1 is coated with polydopamine and amino-modified metal-organic framework (MOF) nanocomposites with zirconium as the central metal ion as a solid-phase microextraction adsorption separation medium for HRP enzymolysis Enzyme solution or β-Casein enzymolysis solution and bovine serum albumin (BSA) enzymolysis solution enrichment and MALDI-TOF MS detection.

[0068] (1) Preparation of standard protein hydrolyzate: Accurately weigh 2 mg of standard protein HRP, 2.5 mg of standard protein β-Casein and 5 mg of standard protein BSA, and use 25 mM ammonium bicarbonate solution to make a concentration of 2 mg / mL, 2.5 mg / mL and 5 mg / mL standard protein solutions, pH about 8.3, boiled for 10 minutes. According to the ratio of trypsin to standard protein at a mass ratio of 1:50, add trypsin and incubate at 37°C for 16 hours to obtain 2 mg / mL HRP trypsin hydrolyzate and 2.5 mg / mL β- Casein enzymatic hydrolysis solution...

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Abstract

The invention provides a synthetic method and application of a metal-organic framework (MOF) composite nanomaterial. The method comprises the following steps: dispersing ferriferrous oxide magnetic spheres which are synthesized through a traditional hydrothermal technology in a weakly alkaline solution of dopamine hydrochloride to carry out self-polymerization of dopamine on the surfaces of the magnetic spheres; and sequentially dispersing polydopamine coated magnetic spheres in a dimethylformamide solution of zirconium chloride and a dimethylformamide solution of 2-amino-terephthalic acid to obtain the MOF composite nanomaterial with the magnetic sphere surfaces coated with polydopamine and modified with an amino group and with zirconium as a center metal ion. The material has the advantages of large specific surface area, good hydrophilicity and suitable pore structure, can be applied to further researches of the proteomics, and can specifically enrich Which can specifically enrich phosphorylated peptide segments and glycopeptides; the synthetic method is simple and quick; and the synthesized material has good hydrophilicity and biocompatibility, and can be used for selectively enriching endogenous phosphorylation peptide segments and glycopeptide in complex biological samples.

Description

technical field [0001] The invention belongs to the field of advanced nanomaterials and nanotechnology, and specifically relates to a synthesis method and application of a metal-organic framework (MOF) nanocomposite material, in particular to a method for phosphorylated peptide enrichment and glycopeptide enrichment with MALDI- Synthesis method and bidirectional application of metal-organic framework (MOF) nanocomposites with polydopamine and amino-modified zirconium-centered metal ions on the surface of magnetic balls detected by TOF MS and LC-MS / MS. Background technique [0002] Protein glycosylation and phosphorylation are two important and common post-translational modifications in the life process. They are closely related to many important complex biological processes such as intercellular signal transmission, cell division, proliferation, differentiation and interaction. Some studies have shown that abnormal expression levels of glycopeptides or phosphorylated peptide...

Claims

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Application Information

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IPC IPC(8): B01J20/26B01J20/28B01J20/30B01D15/08C07K1/22C07K1/16G01N33/68
CPCB01J20/26B01D15/08B01J20/0229B01J20/223B01J20/28009C07K1/16C07K1/22G01N33/6851
Inventor 邓春晖谢伊沁
Owner FUDAN UNIV
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