Preparation and application of a guanidinium-immobilized affinity magnetic sphere with core-shell structure

A technology of core-shell structure and shell structure is applied in the field of preparation and application of guanidine-immobilized affinity magnetic spheres with core-shell structure, which can solve the problems such as the inability to comprehensively and deeply obtain phosphorylated proteomic information, and achieve rapid completely separate effect

Active Publication Date: 2017-09-12
SOOCHOW HIGH TECH CHROMATOGRAPHY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, fixed metal ion affinity chromatography tends to enrich polyphosphopeptides during separation and enrichment, while metal oxide affinity chromatography tends to enrich monophosphopeptides and acidic peptides. Using these two methods alone cannot comprehensively and deeply Obtain phosphoproteomic information

Method used

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  • Preparation and application of a guanidinium-immobilized affinity magnetic sphere with core-shell structure
  • Preparation and application of a guanidinium-immobilized affinity magnetic sphere with core-shell structure
  • Preparation and application of a guanidinium-immobilized affinity magnetic sphere with core-shell structure

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Embodiment 1

[0027] Weigh 300mg of magnetic balls with a diameter of 220nm, disperse them in 50mL of ethanol and aqueous solution with a volume ratio of 4:1, add 1.5mL of ammonia water, sonicate for 10min, and add 0.8mL of 3-(trimethoxysilyl)methacrylic acid dropwise Ester, stirred mechanically at 60°C for 24h to obtain vinyl-modified magnetic spheres.

[0028] Weigh 60 mg of vinyl-modified magnetic spheres and disperse them in 40 mL of acetonitrile, add 140 μL of glycidyl methacrylate, 180 mg of N, N-methylenebisacrylamide and 5 mg of azobisisobutyronitrile, ultrasonicate for 10 min, and place in React at 100°C for 1.5 hours to obtain a polymer-coated magnetic sphere with a core-shell structure, with superparamagnetic Fe3O4 microspheres as the inner core and a polymer layer as the outer shell.

[0029] Disperse 200 mg of polymer-coated magnetic balls in 80 mL of anhydrous ethylenediamine solution, stabilize at 80 ° C, and mechanically stir for 3 h to obtain amino-bonded polymer-coated mag...

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Abstract

The invention relates to the preparation and application of a guanidine-based immobilized affinity magnetic sphere with a core-shell structure, which adopts superparamagnetic iron tetroxide microspheres as a substrate, and 3-(trimethoxysilyl)methanol is bonded on the surface Vinyl-modified magnetic spheres were formed from acrylate-based acrylates, and polymer-coated magnetic spheres with a core-shell structure were obtained by distillation-precipitation polymerization; guanidinium-immobilized core-shell structures were obtained by functional modification of the polymer layer. Polymer coated magnetic balls. In the preparation material of the present invention, utilizing the strong interaction between the guanidinium group and the phosphate group, the polyphosphopeptides in the α-casein (α-casein) or β-casein (α-casein) enzymolysis solution can be respectively realized Or the selective enrichment of all phosphopeptides, the enrichment capacity for phosphopeptides reaches 200 mg / g, the detection limit for phosphopeptides reaches 0.5 fmol, and the recovery rate for phosphopeptides enrichment reaches 91.30%. Utilizing the superparamagnetism of the material, rapid and complete separation can be achieved within 30s.

Description

【Technical field】 [0001] The invention relates to the field of magnetic nanomaterials, the field of separation, analysis and detection of low-abundance special structural proteins or polypeptides, and specifically relates to a method for preparing a polymer with superparamagnetic iron tetroxide microspheres as the core by distillation precipitation polymerization. The layer is a core-shell structure material wrapped in a shell, and the fixed guanidinium group is used to realize the specific enrichment detection method for proteins or polypeptides with special structures. 【Background technique】 [0002] Protein reversible phosphorylation modification is one of the important post-translational modification methods. Although phosphoproteomics has made great progress, there are still some problems, such as low ionization efficiency of phosphopeptides, low abundance phosphopeptide signals often Inhibited and interfered with by the concurrent high-abundance non-phosphopeptide sign...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08F292/00C08F8/32G01N27/62G01N30/02G01N27/626
Inventor 张维冰熊志超赵祎曼邵平戈兆松
Owner SOOCHOW HIGH TECH CHROMATOGRAPHY
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