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Zirconium phosphoester nano magnetic beads, as well as preparation and application thereof

A technology of nano-magnetic beads and phosphate esters, applied in peptide preparation methods, chemical instruments and methods, organic chemistry, etc., can solve problems such as unseen reports, achieve high-selectivity separation and enrichment characteristics, and high-efficiency separation and enrichment , the effect of reducing sample loss

Inactive Publication Date: 2009-05-20
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the method of immobilizing metal zirconium ions on the surface of functionalized nano-magnetic beads and applying them to the separation and enrichment of phosphorylated peptides.

Method used

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  • Zirconium phosphoester nano magnetic beads, as well as preparation and application thereof
  • Zirconium phosphoester nano magnetic beads, as well as preparation and application thereof
  • Zirconium phosphoester nano magnetic beads, as well as preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Enrichment of Phosphorylated Peptides Using Zirconium Phosphate Nanomagnetic Beads

[0031] Preparation of zirconium phosphate nano-magnetic beads:

[0032] 1. Ferric oxide core magnetic beads are dispersed in 30ml of absolute ethanol solution under the action of ultrasound, and 1.0ml of ethyl tetrasilicate (TEOS) is added in a 40°C water bath with stirring. After 2 hours of reaction, use a magnet Absorb the magnetic beads, pour off the supernatant, wash the magnetic beads with absolute ethanol, then add 80ml of absolute ethanol, reflux in a 60°C water bath for 12 hours, attract the magnetic beads with a magnet, and pour off the supernatant.

[0033] 2. After the nano-magnetic beads covered with silica are placed in a vacuum desiccator for 12 hours, add 1.5ml of 3-aminopropyltriethoxysilane (APTES) to 35ml of anhydrous toluene solution, under N2 protection and 110°C Stir and reflux for 10 hours, then use a magnet to hold the magnetic beads, pour off the super...

Embodiment 2

[0044] Preparation of zirconium phosphate nano-magnetic beads:

[0045] 1.0.15g ferric iron tetroxide core magnetic beads are dispersed in 60ml of absolute ethanol solution under the action of ultrasound, add 1.5ml of ethyl tetrasilicate (TEOS) under stirring in a water bath at 45°C, react for 3 hours, and use The magnetic beads are attracted by a magnet, and the supernatant is discarded. After the magnetic beads are washed with absolute ethanol, 90ml of absolute ethanol is added, and refluxed in a water bath at 45°C for 13 hours. The magnetic beads are attracted by a magnet, and the supernatant is discarded. .

[0046] 2. After the nano-magnetic beads covered with silica were placed in a vacuum desiccator for 8 hours, 1.5ml of 3-aminopropyltriethoxysilane (APTES) was added to 20ml of anhydrous toluene solution. 2 Protect and reflux at 90°C for 4 hours, then hold the magnetic beads with a magnet, pour off the supernatant to obtain aminated nano-magnetic beads, wash them with ...

Embodiment 3

[0050] Preparation of zirconium phosphate nano-magnetic beads:

[0051] 1.0.25g ferric oxide core magnetic beads are dispersed in 80ml of absolute ethanol solution under the action of ultrasound, under 50°C water bath and stirring, add 2.0ml ethyl tetrasilicate (TEOS), react for 4 hours, use The magnetic beads are attracted by a magnet, and the supernatant is discarded. After the magnetic beads are washed with absolute ethanol, 100ml of absolute ethanol is added, and refluxed in a water bath at 65°C for 14 hours. The magnetic beads are attracted by a magnet, and the supernatant is discarded. .

[0052] 2. After the nano-magnetic beads covered with silica were placed in a vacuum desiccator for 16 hours, 1.5ml of 3-aminopropyltriethoxysilane (APTES) was added to 40ml of anhydrous toluene solution. 2 Protect and reflux at 115°C for 8 hours, then use a magnet to hold the magnetic beads, pour off the supernatant to obtain aminated nano-magnetic beads, wash them with toluene, disti...

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Abstract

The invention relates to a phosphate zirconium nano magnetic particle with the structural representation on the right, and the preparation and application thereof; the preparation method comprises the steps that phosphate groups are induced to the surface of the nano magnetic particle which is wrapped with silicon dioxide on the surface and takes ferriferrous oxide as a magnetic inner core; then, a phosphate zirconium layer is formed on the surface by the mutual action between zirconium ions and the phosphate groups on the surface, thereby successively preparing a nano magnetic particle having high efficiency, high-selectivity separation and enrichment feature to peptide phosphate. The nano magnetic particle is mainly used for separating the enriched peptide phosphate from biological samples, with high efficiency and under the action of a magnetic field. The extracted peptide phosphate can be directly analyzed and represented by mass spectrum.

Description

technical field [0001] The invention relates to the separation and enrichment of phosphorylated peptides, in particular to zirconium phosphate nanometer magnetic beads and its preparation, as well as its application in the selective separation and enrichment of phosphorylated peptides. Background technique [0002] Protein separation analysis is a research hotspot in the field of biological separation analysis in recent years. Protein post-translational modification is an important factor affecting protein structure and function, and plays an important role in the regulation of biochemical pathways, among which protein phosphorylation is the most common It is also the most important post-translational modification method, which is a hot spot and difficulty in proteomics research. Protein phosphorylation and dephosphorylation regulate almost the entire process of human life activities, including cell proliferation, development and differentiation, neural activity, muscle cont...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/14
Inventor 邹汉法赵樑吴仁安周厚江
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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