Sea-urchin-shaped core-shell type Fe3O4@TiO2 magnetic microspheres, and preparation and application thereof

A magnetic microsphere and sea urchin-shaped technology, which is applied to the magnetic properties of inorganic materials, the preparation of test samples, and other chemical processes, can solve the problem of insufficient surface affinity sites, unsatisfactory enrichment effect of phosphorylated peptides, and comparative Small surface area and other issues, to achieve high selectivity and enrichment capacity, good practical prospects and application value, simple processing effect

Active Publication Date: 2014-12-10
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, the existing metal oxide magnetic materials have a small specific surface area and insufficient surface affinity sites, resulting in unsatisfactory enrichment of phosphorylated peptides

Method used

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  • Sea-urchin-shaped core-shell type Fe3O4@TiO2 magnetic microspheres, and preparation and application thereof
  • Sea-urchin-shaped core-shell type Fe3O4@TiO2 magnetic microspheres, and preparation and application thereof
  • Sea-urchin-shaped core-shell type Fe3O4@TiO2 magnetic microspheres, and preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1. Sea urchin-shell type Synthesis of magnetic materials.

[0022] The whole synthesis reaction is divided into four steps:

[0023] In the first step, Fe was synthesized by solvothermal reduction 3 o 4 Superparamagnetic microspheres: 1.35gFeCl 3 ·6H 2O was added to 40 mL of ethylene glycol and stirred until the solution was clear. Add 3.6g of sodium acetate and 1g of polyethylene glycol (4000-6000), stir well until the solution is uniform. The solution was transferred to a 50mL reactor, the temperature was raised to 200°C, and the reaction was carried out for 8 hours. After natural cooling, ultrasonically clean with water and ethanol several times. The final product was dried at 60°C for further use.

[0024] In the second step, the sol-gel method is used to synthesize the Magnetic microspheres: firstly, the Fe obtained from step 1 3 o 4 30 mg of magnetic microspheres were redispersed in 40 mL of 1:4 water / ethanol solution under ultrasonic conditio...

Embodiment 2

[0028] Example 2. Selective enrichment of phosphorylated polypeptides in a complex polypeptide mixture and matrix-assisted laser desorption ionization mass spectrometry analysis.

[0029] (1) Preparation of α-casein protein hydrolyzate. α-casein was dissolved in an ammonium bicarbonate buffer solution with pH=8 to form a 1mg / ml solution, and trypsin was added in a proportion of 2.5% by weight, and the enzymolysis was carried out at 37°C for 18 hours, and the obtained polypeptides were mixed The solution was diluted with 0.1% trifluoroacetic acid aqueous solution containing 50% acetonitrile to a concentration of 4×10 -7 mol / L.

[0030] (2) Selective enrichment and MALDI-TOF / MS analysis of phosphorylated peptides. 5 μL 10mg / mL of the sea urchin-shaped core-shell type prepared in Example 1 The microsphere aqueous solution was added to 200 μL with a concentration of 4×10 -7 mol / L α-casein trypsin hydrolyzed mixed polypeptide solution, vortexed for 3 minutes to capture phospho...

Embodiment 3

[0034] Example 3 Adjust the concentration of α-casein trypsin hydrolysis mixed polypeptide solution to 2×10 -9 mol / L, other conditions are the same as example 2, the result is as Figure 5 (a) and (b), it can be seen from the figure that even if the concentration of phosphorylated polypeptide drops to the pmol / L level, there are still phosphorylated polypeptide peaks that can be detected, indicating that the sea urchin-shaped core-shell material has a high collection capacity.

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Abstract

The invention belongs to the technical field of inorganic material and analysis, and specifically relates to sea-urchin-shaped core-shell type magnetic microspheres, preparation method and application thereof in the respect of phosphorylated polypeptide selective enrichment. According to the magnetic microspheres, a layer of a SiO2 material is coated on the surface of Fe3O4 magnetic microspheres with a sol-gel method; a layer of anatase nuclei is deposited; and through hydrothermal growth, rutile nano-rods are grown on the surface of the nano-grade nuclei, such that the magnetic microsphere particles with the sea urchin shapes are formed. The magnetic material has large specific surface area, and rapid external magnetic field response. During selective enrichment of trace amount of phosphorylated polypeptide in a biological sample, the treatment is simple, and specificity is high. The material has good practical value and application prospect in the fields such as proteomics.

Description

technical field [0001] The invention belongs to the technical field of inorganic materials and analysis, in particular to a sea urchin-shaped core-shell type Magnetic microspheres and their preparation methods and applications. [0002] The magnetic microspheres proposed in the present invention can be used as an affinity probe to achieve highly selective enrichment of trace phosphorylated peptides in biological samples containing complex peptides, and combined with MALDI-TOF / MS to enrich the phosphorylated Peptides were analyzed and identified. The material has good practical value and application prospect in proteomics and other fields. Background technique [0003] Protein phosphorylation is one of the important ways of protein post-translational modification. Protein phosphorylation is a common phenomenon in cells. These phosphorylated proteins play an important role in cell signal transduction, cell differentiation and cell growth and other life processes. Regulator...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/08B01J20/30H01F1/01G01N1/40
Inventor 许国旺李华石先哲单圆鸿
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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