Preparation method of core-shell type boryl magnetic microsphere capable of greatly enriching glycoprotein

A technology of magnetic microspheres and glycoproteins, which is applied in the preparation of microspheres and microcapsule preparations. It can solve the problems of poor control of structure and magnetic content, and achieve the effects of controllable magnetic content, excellent effect, and uniform particle size distribution.

Active Publication Date: 2019-01-18
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 type boryl magnetic microsphere capable of greatly enriching glycoprotein
  • Preparation method of core-shell type boryl magnetic microsphere capable of greatly enriching glycoprotein
  • Preparation method of core-shell type boryl magnetic microsphere capable of greatly enriching glycoprotein

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 preparation method of a core-shell type boryl magnetic microsphere capable of greatly enriching glycoprotein. The preparation method comprises the following steps: taking ferric trichloride hexahydrate and acetate as raw materials, adopting a solvothermal method to prepare nano ferroferric oxide magnetic particles; adopting a sol-gel method to surface a layer of SiO2 on the surfaces of the nano ferroferric oxide magnetic particles; modifying the magnetic microsphere coated with SiO2, and enabling the surface of the magnetic microsphere to obtain an active vinyl functional group; preparing the core-shell type boryl magnetic microsphere with amino rich in surface through polymerization reaction; modifying a great number of borate groups by the amino in the surface and carboxybenzene borate through amidation reaction, thereby obtaining the core-shell type boryl magnetic microsphere. The core-shell type boryl magnetic microsphere prepared by the preparation methodis uniform in particle diameter distribution, is regular in structure, is controllable in magnetic content, has the characteristics of high magnetic response and surface modification, can separate and purify glycoprotein under a physiological condition, is good in separating ability, and is a biological magnetic separating material with application prospect.

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 Applications(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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