Synthetic method and application of metal organic skeleton-modified magnetic grapheme composite material

A metal-organic framework and magnetic graphene technology, applied in the field of nanomaterials, can solve the problems of weak ionization ability and low abundance of glycosylated proteins/peptides, and achieve good hydrophilic pore structure, high repetition rate, high reliability and high reliability. high noise effect

Inactive Publication Date: 2016-11-23
FUDAN UNIV
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  • Summary
  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

In recent years, biological mass spectrometry has been widely used in the study of protein post-translational modification, but due to the weak ionization ability and low abundance of glycosylated proteins / peptides, and the mass spectrometry signals of glycosylated peptides are often Masked by ylated peptides, salts, and impurity molecules, it will be a huge challenge to directly use mass spectrometry for analysis and detection

Method used

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  • Synthetic method and application of metal organic skeleton-modified magnetic grapheme composite material
  • Synthetic method and application of metal organic skeleton-modified magnetic grapheme composite material
  • Synthetic method and application of metal organic skeleton-modified magnetic grapheme composite material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: A synthetic method of a metal-organic framework-modified magnetic graphene composite.

[0032] (1) Disperse 400mg of graphene into 60mL of concentrated nitric acid, and reflux in a water bath at 60°C for 8 hours;

[0033] (2) centrifuging and separating the product obtained in step (1), washing it with deionized water until the solution is neutral, drying it in vacuum at 50°C, placing it for storage, and obtaining acidified graphene;

[0034] (3) 405mg FeCl 3 ·6H 2 O was dissolved in 40mL ethylene glycol solution, and stirred to obtain a yellow transparent solution;

[0035] (4) Disperse 150 mg of the product obtained in step (2) in the solution obtained in step (3), and sonicate for 1.0 hour;

[0036] (5) Under ultrasonic stirring, add 150 mg of trisodium citrate, 1.8 g of sodium acetate and 1.0 g of polyethylene glycol (20000) into the mixed solution obtained in step (4), and fully stir for 1.0 hour;

[0037](6) Transfer the mixed solution obtained in s...

Embodiment 2

[0045] Example 2: The above metal-organic framework-modified magnetic graphene composite material was applied to the separation, enrichment and MALDI-TOF-MS detection of glycosylated peptides in the standard HRP enzymatic hydrolysis solution.

[0046] (1) Preparation of standard HRP protein enzymatic solution: Accurately weigh 1mg of standard protein HRP and dissolve in 25mM ammonium bicarbonate buffer, boil for 8 minutes, dilute to 1mg / mL with 25mM ammonium bicarbonate buffer, and then Add an appropriate amount of trypsin at a ratio of 1:40, and digest overnight at 37°C for 16 hours;

[0047] (2) Wash 5 mg of the metal-organic framework-modified magnetic graphene composite with 85% acetonitrile / 1.0% trifluoroacetic acid buffer three times, and then disperse in 500 μL of 85% acetonitrile / 1.0% trifluoroacetic acid buffer , ultrasonically disperse until uniform, and prepare a 10mg / mL solution;

[0048] (3) Enrichment of glycosylated peptides: take 50 μL of the solution obtained...

Embodiment 3

[0051] Example 3: The magnetic graphene composite material modified by a metal organic framework obtained in Example 1 is applied to the enrichment and MALDI-TOF-MS of HRP enzymolysis solution and HRP protein and bovine serum albumin (BSA) protein mixed solution detection.

[0052] (1) Mix the HRP enzymatic solution with the HRP and BSA protein solution according to the protein mass ratio of 1:800:800, take 2 μL of the standard mixed solution and add it to 150 μL of 85% acetonitrile / 1.0% trifluoroacetic acid buffer solution, and then add 50 μL of The dispersion of metal-organic framework-modified magnetic graphene composites (concentration: 10 mg / mL) was incubated and swirled at 37°C for 30 minutes; the material was separated by centrifugation and washed three times with 200 μL of 85% acetonitrile / 1.0% trifluoroacetic acid buffer Afterwards, elute with 10 μL 30% acetonitrile / 0.1% trifluoroacetic acid buffer for 15 minutes, and magnetically separate;

[0053] (2) Spotting the ...

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Abstract

The invention belongs to the technical field of nanometer materials and particularly relates to a metal organic skeleton-modified magnetic grapheme composite material as well as a synthetic method and application thereof. The method comprises the following steps: synthesizing a magnetic grapheme composite material by utilizing a one-pot method, dispersing magnetic grapheme into an aqueous solution of zinc nitra tehexahydrate and 2-methylimidazole and performing ultrasonic reaction at the room temperature to prepare the metal organic skeleton-modified magnetic grapheme composite material taking zinc ions as the center. Hydrophilic active sites on a coating are densely distributed on the surface of the magnetic graphene material and efficiently react with a hydrophilic part on a glycosylated peptide molecule, so that a target molecule is firmly bonded to the surface of the material. An experiment shows that the material is strong in hydrophilicity, large in specific surface area and stable in biocompatibility; when being utilized for immobilizing glycosylated peptides, the composite material has the advantages of being high in reaction efficiency, small in nonspecific adsorption and the like and realizes efficient separation and enrichment of low-abundance glycosylated peptide mixed samples. The metal organic skeleton-modified magnetic grapheme composite material provided by the invention is low in cost, practical and efficient, high in stability, good in repeatability and huge in application prospect.

Description

technical field [0001] The invention belongs to the technical field of nanometer materials, and in particular relates to a metal-organic framework-modified magnetic graphene composite material and a synthesis method and application thereof. Background technique [0002] Proteins are the direct embodiment of life phenomena, and the study of protein functions will help to understand the physiological and pathological mechanisms of life activities. Protein glycosylation is the most important and common post-translational modification of proteins in biological cells, and it plays an irreplaceable regulatory role in metabolism, signal transduction, cell proliferation and division, gene expression and other processes in organisms. In recent years, biological mass spectrometry has been widely used in the study of protein post-translational modification, but due to the weak ionization ability and low abundance of glycosylated proteins / peptides, and the mass spectrometry signals of g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/22B01J20/28B01J20/30
CPCB01J20/226B01J20/20B01J20/28009B01J2220/46
Inventor 高明霞王嘉希张祥民李杰
Owner FUDAN UNIV
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