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Preparation method of core-shell borylated magnetic microspheres capable of enriching glycoproteins in large quantities

A technology of magnetic microspheres and glycoproteins, applied in the preparation of microspheres, microcapsule preparations, etc., can solve the problems of poor control of structure and magnetic content, and achieve the effects of controllable magnetic content, high magnetic response and excellent effect

Active Publication Date: 2021-08-06
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, emulsion polymerization has relatively poor control over the structure and magnetic content of the prepared microspheres.

Method used

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  • Preparation method of core-shell borylated magnetic microspheres capable of enriching glycoproteins in large quantities
  • Preparation method of core-shell borylated magnetic microspheres capable of enriching glycoproteins in large quantities
  • Preparation method of core-shell borylated magnetic microspheres capable of enriching glycoproteins in large quantities

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Preparation of core-shell iron ferric oxide@silica@polypropyleneamine-carboxyphenyl borate magnetic composite microspheres with a shell thickness of about 25nm and a particle size of about 250nm

[0030] The specific steps are:

[0031] (1) Preparation of magnetic Fe3O4 nanoparticles

[0032] Dissolve 2g of ferric chloride hexahydrate and 4g of sodium acetate in 10mL of ethylene glycol / ethylenediamine (2:1, V / V) mixed solvent, stir mechanically at room temperature for 15 minutes, then transfer the solution into In a tetrafluoroethylene-lined stainless steel high-temperature and high-pressure reaction kettle, place the reaction kettle in an oven at 200°C for 20 hours, and cool to room temperature naturally; use a magnet to separate the black sample, and then wash the black substance with absolute ethanol 4 times , washed with purified water three times, separated the sample with a magnet, and dried in a vacuum oven at 30-50°C for 12 hours. The dried magnetic ...

Embodiment 2

[0041] Example 2 Preparation of core-shell iron ferric oxide@silica@polypropyleneamine-carboxyphenyl borate magnetic composite microspheres with a shell thickness of about 40nm and a particle size of about 300nm

[0042] The specific steps are:

[0043] (1) Preparation of magnetic Fe3O4 nanoparticles

[0044] With embodiment 1.

[0045] (2) Preparation of ferroferric oxide@silicon dioxide nano-magnetic composite particles

[0046] With embodiment 1 step.

[0047] (3) Preparation of vinyl-modified ferric oxide@silica nano-magnetic composite particles

[0048] With embodiment 1.

[0049] (4) Preparation of Fe3O4@Silicon Dioxide@Polypropyleneamine Nanomagnetic Composite Particles

[0050] Wherein the consumption of allylamine is 1mL; Others are the same as in Example 1.

[0051] (5) Preparation of ferroferric oxide@silica@polyacrylamine-carboxyphenyl borate magnetic composite microspheres

[0052] With embodiment 1.

[0053] figure 1 , figure 2 The transmission electro...

Embodiment 3

[0054] Example 3 Using the magnetic composite microspheres prepared in Example 2 to conduct an enrichment experiment on the isolated glycoprotein horseradish peroxidase (HRP) in a complex fetal bovine serum system

[0055] The specific method is:

[0056] (1) First, weigh 1 mg ferric oxide@silica@polyacrylamine-carboxyphenyl borate magnetic composite microspheres, and wash twice with 100 μL buffer solution Ⅰ (10mM PBS, pH=7.4);

[0057] (2), then add 5 μL (40 μL / mL) mixture of glycoprotein and 5 μg glycoprotein HRP, add buffer solution I to 100 μL, incubate at room temperature for 10 minutes;

[0058] (3) Collect the supernatant by magnetic separation, add 100 μL of buffer solution I to wash twice;

[0059] (4) Finally, elute with 50 μL buffer solution II (50% AN containing 1% TFA), take 10 μL stock solution, 10 μL supernatant and 10 μL eluate, add 10 μL bromophenol blue loading buffer after freeze-drying, and run electrophoresis , the electropherogram of which is shown in ...

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Abstract

The invention discloses a method for preparing core-shell type borylated magnetic microspheres capable of enriching a large amount of glycoproteins. Firstly, ferric trichloride hexahydrate and acetate are used as raw materials, and nanometer trioxide tetroxide is prepared by a solvothermal method. Ferromagnetic particles; the surface of nano ferromagnetic particles is coated with a layer of SiO by sol-gel method 2 ; For coating with SiO 2 Modified the magnetic microspheres to make the surface contain active vinyl functional groups; prepare the core-shell magnetic composite microspheres with rich amino groups on the surface by polymerization reaction; boronate groups to obtain core-shell borylated magnetic microspheres. The core-shell type borylated magnetic microspheres prepared by the present invention have uniform particle size distribution, regular structure, controllable magnetic content, high magnetic responsiveness and surface modifiable characteristics, and can separate and purify glycoproteins under physiological conditions. The separation ability is good, and it is a biomagnetic separation material with great application prospects.

Description

technical field [0001] The invention relates to the preparation of magnetic polymer microspheres, in particular to a method for preparing core-shell type borylated magnetic microspheres capable of enriching glycoproteins in large quantities in the purification and separation of glycoproteins. Background technique [0002] Glycoproteins are functional proteins that have carbohydrates covalently attached to protein side chains. As one of the most important post-translational modifications of proteins in organisms, glycosylation plays an important role in many physiological regulatory processes such as cell recognition, signal transduction, immune protection, and inflammation. Glycoprotein research has important biological significance and potential clinical applications. Mass spectrometry has been introduced into the study of glycoproteins to obtain detailed information about them, especially with regard to glycosylation site occupancy at each glycol site and glycan heterogen...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J13/14
CPCB01J13/14
Inventor 朱飞飞杨书彬马建军陈飞李林渠海郑业焕付光宇吴学炜
Owner AUTOBIO DIAGNOSTICS CO LTD
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