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Synthesis method and application of GO@CS@ZIF-8 foam material

A technology of foam material and synthesis method, which is applied in the synthesis and application field of GO@CS@ZIF-8 foam material, can solve the problems of poor glycopeptide enrichment effect, limited application, inability to realize phosphorylated peptide enrichment, etc., and achieve The effect of high selectivity, accurate determination, and good enrichment and purification ability

Active Publication Date: 2021-11-30
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] As a member of the MOF family, ZIF-8 not only has the advantages of MOF, but also has the advantages of simple synthesis, easy modification and controllable assembly. application, and pure ZIF-8 particles have poor enrichment effect on glycopeptides due to the limitation of their own properties, and their application in actual samples is limited, while pure hydrophilic materials cannot achieve the enrichment of phosphorylated peptides

Method used

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  • Synthesis method and application of GO@CS@ZIF-8 foam material
  • Synthesis method and application of GO@CS@ZIF-8 foam material
  • Synthesis method and application of GO@CS@ZIF-8 foam material

Examples

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preparation example Construction

[0042] A kind of synthetic method of GO@CS@ZIF-8 foam material, comprises the steps:

[0043] Step 1: Dissolve graphene oxide in 1% acetic acid solution and ultrasonically disperse for half an hour. Graphene oxide contains hydroxyl, carboxyl and epoxy functional groups, which can be combined with chitosan through hydrogen bond and amide reaction, and then passed through the ice template to form a 3D structure.

[0044] Then add chitosan, 25% glutaraldehyde aqueous solution successively under stirring condition, stir 12 hours, it is carried out freeze-drying after completion of reaction; The present invention adds chitosan because there is a large amount of amino groups on its surface, can strengthen the base material Hydrophilic, thereby improving the enrichment effect on glycopeptides. Too much addition of chitosan will make the bond between graphene oxide and chitosan too strong, which is not conducive to the dispersion in the later stage, and too little addition will lead ...

Embodiment 1

[0057] A synthesis method of GO@CS@ZIF-8 foam material, including: Step 1: Dissolve 10mg graphene oxide in 1% acetic acid solution, ultrasonically disperse for half an hour, add 40mg chitosan, 24μL 25% glutaraldehyde aqueous solution was stirred for 12 hours, and freeze-dried after the reaction was completed.

[0058] Step 2: adding the product obtained in step 1 to a methanol solution of zinc nitrate hexahydrate, stirring for 10 minutes, adding a methanol solution of 2-methylimidazole, and stirring the resulting mixed solution at room temperature for 4 hours;

[0059] Step 3: The product obtained in step 2 was fully washed with pure methanol and then dried under vacuum at 60°C for 12 hours to synthesize GO@CS@ZIF-8 foam material.

[0060] The prepared GO@CS@ZIF-8 foam material was detected with a high-resolution scanning electron microscope. The model of the high-resolution scanning electron microscope was JEM2100. The GO@CS@ZIF-8 foam material was evenly spread on the conduc...

Embodiment 2

[0064] The GO@CS@ZIF-8 foam material prepared in Example 1 was used as a fixed extraction agent for the separation of glycosylated peptides from the peptide mixture enzymatically hydrolyzed by standard glycoprotein horseradish peroxidase (HRP). Set purification.

[0065] Step 1: Dissolve 1 mg HRP in 1 mL 50 mM NH 4 HCO 3 Enzymolysis in the solution for 16 hours at a temperature of 37°C to obtain the HRP enzymolysis solution containing glycopeptides;

[0066] Step 2: Dissolve 500 μg GO@CS@ZIF-8 foam material with 200 μL buffer ACN / H 2 O / TFA (90 / 9 / 1, v / v / v) prepared as a dispersion;

[0067] Step 3: Take 6 μL 663 fmol of the HRP hydrolyzate prepared in Step 1 and add it to the dispersion prepared in Step 2, mix and enrich at room temperature for 30 minutes;

[0068] Step 4: Centrifuge the enriched dispersion from step 3, remove the supernatant, and replace with 200 μL of buffer ACN / H from step 2 2 O / TFA (90 / 9 / 1, v / v / v) washed three times;

[0069] Step 5: Then with 10 μL e...

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Abstract

The invention relates to a synthesis method and application of a GO@CS@ZIF-8 foam material. The preparation method comprises the following steps of: 1, dissolving graphene oxide in a solvent, carrying out ultrasonic dispersion, adding chitosan and a glutaraldehyde aqueous solution while performing stirring, and carrying out freeze drying after reaction; and 2, adding a product in the step 1 into a methanol solution of zinc nitrate hexahydrate, adding a methanol solution of 2-methylimidazole while stirring, and carrying out a stirring reaction on the obtained mixed solution at room temperature to obtain the GO@CS@ZIF-8 foam material. The invention relates to a glycopeptide and phosphorylated peptide enrichment and mass spectrometric detection method. The method comprises the following steps of: 1, dissolving target protein in an NH4HCO3 solution for enzymolysis for 16 hours; 2, preparing a dispersion liquid from the GO@CS@ZIF-8 foam material by using a buffer solution; 3, adding target peptide into the dispersion liquid, performing mixing, oscillating and enriching; 4, centrifuging the dispersion liquid, removing supernate, and performing washing with the buffer solution; 5, eluting the target peptide with eluent; and 6, carrying out mass spectrometry on the eluent subjected to elution.

Description

technical field [0001] The invention belongs to the preparation of a novel porous foam base material and its application in simultaneous glycopeptide and phosphorylated peptide, and specifically relates to a synthesis and application method of a GO@CS@ZIF-8 foam material. Background technique [0002] Glycosylation and phosphorylation of proteins are two of the most important post-translational modifications of proteins, which are involved in many life activities, such as intercellular reactions, signal transduction and metabolism. Therefore, abnormal post-translational modifications will lead to changes in protein structure and function, and will change the role of proteins and other molecules, thereby causing changes in biological processes. For example, currently incurable cancers are related to abnormal protein glycosylation and phosphorylation modifications , and more and more studies have shown that cancer is related to abnormal protein post-translational modification....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68B01J20/28B01J20/282B01J20/22B01J20/20B01J20/24
CPCG01N33/6848B01J20/28045B01J20/282B01J20/226B01J20/20B01J20/24
Inventor 刘容高文清杨佳倩李俊晖俞建成唐科奇
Owner NINGBO UNIV
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