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

A magnetic microsphere, sea urchin-like technology, applied in the fields of inorganic material magnetism, preparation of test samples, other chemical processes, etc., can solve the problem that the enrichment effect of phosphorylated peptides is not ideal, the specific surface area is small, and the surface affinity Insufficient points and other problems, to achieve the effect of good practical prospects and application value, simple processing process, high selectivity and enrichment ability

Active Publication Date: 2013-06-19
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-shaped core-shell type Synthesis of magnetic materials.

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

[0023] The first step is to synthesize Fe by solvothermal reduction method 3 O 4 Superparamagnetic microspheres: 1.35gFeCl 3 ·6H 2 O 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) and 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, ultrasonic cleaning with water and ethanol several times. The final product was dried at 60°C for use.

[0024] The second step is to synthesize a core-shell structure by sol-gel method Magnetic microspheres: firstly, the Fe prepared by step 1 3 O 4 30 mg of magnetic microspheres were re-dispersed in 40 mL of 1:4 water / ethanol solution under ultrasonic con...

Embodiment 2

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

[0029] (1) Preparation of α-casein proteolysis solution. α-casein is dissolved in ammonium bicarbonate buffer solution with pH=8 to form a 1mg / ml solution, trypsin is added at a weight fraction of 2.5%, enzymatic hydrolysis is carried out at 37°C for 18 hours and then terminated, the resulting polypeptide is mixed The solution is diluted with a 0.1% trifluoroacetic acid aqueous solution containing 50% acetonitrile to a concentration of 4×10 -7 mol / L.

[0030] (2) Selective enrichment of phosphorylated peptides and MALDI-TOF / MS analysis. 5μL 10mg / mL sea urchin-shaped core-shell prepared in Example 1 The aqueous solution of microspheres is added to 200μL at a concentration of 4×10 -7 In the mol / L α-casein tryptic peptide solution, vortex for 3 minutes to capture the phosphorylated peptide. Afterwards, th...

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 in Example 2, the results are as follows Figure 5 (a) and (b), the figures show that even if the concentration of phosphorylated peptide drops to pmol / L, there are still phosphorylated peptide peaks that can be detected, indicating that the sea urchin-like core-shell type material is highly enriched. Set 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 Fe3O4@TiO2 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, and specifically relates to a sea urchin-like core-shell type Magnetic microspheres and preparation methods and applications thereof. [0002] The magnetic microspheres proposed by the present invention can be used as an affinity probe to achieve highly selective enrichment of trace phosphorylated peptides in biological samples containing complex peptide fragments, and combine with MALDI-TOF / MS for the enriched phosphorylation Peptides are analyzed and identified. The material has good practical value and application prospects in proteomics and other fields. Background technique [0003] Protein phosphorylation is one of the important ways of protein post-translational modification. The phosphorylation of proteins in cells is a common phenomenon. These phosphorylated proteins play an important role in cell signal transduction, cell differentiation and cell growth and other lif...

Claims

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Application Information

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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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