Dendrimer-modified MOF material, and preparation method and application thereof

A MOF, dendritic technology, applied in separation methods, chemical instruments and methods, other chemical processes, etc., can solve the problems of single, tedious experimental steps, and low enrichment efficiency.

Inactive Publication Date: 2016-03-09
FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

And each of these methods also has its own advantages and disadvantages. The boric acid method can enrich N-sugar and O-sugar at the same time, but it has strict pH requirements, and the enrichment efficiency is low; the hydrazine chemical method requires very tedious experiments. steps, and the same lectin can only enrich the same type of sugar chains

Method used

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  • Dendrimer-modified MOF material, and preparation method and application thereof
  • Dendrimer-modified MOF material, and preparation method and application thereof
  • Dendrimer-modified MOF material, and preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0019] Example 1: NH 2 -Synthesis of Mil-101(Cr)PAMAM material

[0020] (1) NH 2 -Synthesis of Mil-101(Cr): 500mg chromium nitrate nonahydrate [Cr(NO 3 ) 3 9H 2 O] and 230 mg of 2-aminoterephthalic acid were dissolved in 7 mL of deionized water; after stirring at room temperature for 2 hours, the mixture was transferred to a stainless steel autoclave, reacted at 130 °C for 24 hours, and cooled to room temperature; the cooled solution was centrifuged to obtain a green The precipitate was washed five times with ethanol, and dried in a vacuum oven at 50°C to obtain NH 2 -Mil-101(Cr).

[0021] (2) NH 2 -Mil-101 ​​(Cr) surface grafted dendrimer polyamide-amine (PAMAM): take dry NH 2 -Mil-101(Cr) 2mg in 1.5mLEP tube, add 5% glutaraldehyde aqueous solution; after activation at 37℃ for 2h, centrifuge and wash three times, add 200μL deionized water and 5μLPAMAM; after reacting at 37℃ for 4h, continue to centrifuge and wash Three times, add 150 μL of 10 mg / mL sodium cyanoborohyd...

Embodiment 2

[0022] Embodiment 2: NH2-Mil-101 ​​(Cr) PAMAM material is used for the enrichment of glycopeptide

[0023] Take 500μg NH2-Mil-101(Cr)PAMAM and wash it three times with 95% acetonitrile+1%TFA solution. Add 95% acetonitrile + 1% TFA solution and an appropriate amount of enzymatically hydrolyzed glycopeptide mixture to make the total volume 100 μL. Enrichment was carried out at 37°C for 50 min. The supernatant was separated and removed, and then washed three times with 95% acetonitrile+1% TFA solution. Add 10 μL of eluent (30% ACN, 0.1% TFA) and elute at 37°C for 20 min to elute the glycopeptide. The eluate is analyzed and detected by MALDI-TOFMS.

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Abstract

The invention specifically relates to a dendrimer-modified MOF material and a preparation method thereof, and application of the dendrimer-modified MOF material in enrichment of glycopeptides. The preparation method comprises the following steps: synthesizing a good-stability aminated MOF material of Mil-101(Cr) by using a hydrothermal method; and then grafting hydrophilic dendrimer polyamide-amine (PAMAM) on the surface of MOF by using a glutaraldehyde crosslinking method so as to obtain the dendrimer-modified MOF material. The dendrimer PAMAM for modification has a long chain and a great number of amino groups, so hydrophilicity of the dendrimer-modified MOF material is greatly improved, and the dendrimer-modified MOF material can improve enrichment efficiency when applied to enrichment of glycopeptides. In enrichment of a standard glycoprotein enzymolysis peptide fragment horseradish peroxidase (HPR), 13 glycopeptides is successfully enriched, and the dendrimer-modified MOF material has selectivity of 1: 100 and a detection limit as low as 1 fmol / [mu]L. The dendrimer-modified MOF material is novel, convenient, practical and highly efficient and has good repeatability, high stability and wide application prospects.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a dendrimer-modified MOF material, a preparation method thereof and an application in glycopeptide enrichment. technical background [0002] Protein glycosylation is a very important post-translational modification, such as protein folding, intracellular sorting, recognition and transport and other functions. In recent studies, biological mass spectrometry has been successfully applied to the analysis of glycoproteins as an effective method. However, since glycoproteins are basically low-abundance expressed proteins and the microscopic heterogeneity of sugar chains, at the same time, the chemical nature of sugar chains is polyhydroxy aldehydes, polyhydroxy ketones and their derivatives, lacking functional groups that can produce ultraviolet absorption And because of its strong hydrophilicity, the response signal in mass spectrometry is low. Therefore, it is very importa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/22B01J20/30B01D15/10
Inventor 高明霞张祥民王亚楠王嘉希
Owner FUDAN UNIV
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