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Magnetic microspheres for coupling primary amino-containing bioligands and preparation method thereof

A technology of magnetic microspheres and biological ligands, which is applied to the magnetism of organic materials/organic magnetic materials, silicon dioxide, silicon oxide, etc., can solve the problems of damaging the activity of biological ligands, inconvenient use, cumbersome operation, etc., and achieve Increased binding capacity, easy operation, and simplified coupling process

Active Publication Date: 2016-03-16
BEAVERNANO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the process of coupling bioligands with amino magnetic microspheres, it is also necessary to activate the amino groups with glutaraldehyde in advance, and then covalently couple with the amino groups on the bioligands through aldehyde groups, just like carboxyl magnetic microspheres, Its operation is cumbersome and inconvenient to use, and the introduction of glutaraldehyde may damage the activity of some biological ligands

Method used

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  • Magnetic microspheres for coupling primary amino-containing bioligands and preparation method thereof
  • Magnetic microspheres for coupling primary amino-containing bioligands and preparation method thereof
  • Magnetic microspheres for coupling primary amino-containing bioligands and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Preparation of Carboxylated Magnetic Microspheres Using Silica Magnetic Microspheres as Substrate

[0039] Take 1g of silica magnetic microspheres, disperse them in 300mL of isopropanol solvent, then add 0.2mL of silane coupling agent γ-aminopropyl-triethoxysilane, under the condition of 80℃ water bath, mechanically stir the reaction for 3h, end reaction. The product is separated by magnetic suction, the liquid is removed, and finally washed several times with ethanol to obtain magnetic microspheres whose surface is modified with amino groups. Take 1 g of amino-modified magnetic microspheres and disperse them in 0.1 M NaCl aqueous solution, add 6.4 g of succinic anhydride solid, react on a shaking table at room temperature for 5 h, and end the reaction. The product is separated by magnetic suction to remove the liquid, and finally washed several times with 1M NaCl aqueous solution and deionized water to obtain carboxyl magnetic microspheres with magnetic microsphere bo...

Embodiment 2

[0042] Preparation of Carboxylated Magnetic Microspheres Using Magnetic Agarose Microspheres as Substrate

[0043] Suction filter the magnetic agarose microspheres in a sand core filter funnel, weigh 1 g of the magnetic agarose microspheres after suction filtration and disperse them in acetone solvent, add 3 mL of carbonyldiimidazole (CDI) acetone solution, and mix vertically at room temperature for 3 h, End the reaction. The obtained product was separated by magnetic suction to remove liquid, and washed several times with acetone to obtain CDI-activated magnetic agarose microspheres. Take 1 g of CDI-activated magnetic agarose microspheres and disperse them in 10 mL of deionized water, then weigh the solid glycine and add it to make the final concentration 1 mol / L, finally adjust the pH value to 9.0-10.0, react at room temperature for 16 hours, and end reaction. The obtained solid was separated and removed by magnetic suction, and washed with deionized water, 2M NaCl, and de...

Embodiment 3

[0046] Preparation of Carboxylated Magnetic Microspheres Using Magnetic Microspheres Containing Epoxy Groups on the Surface

[0047] Take 1 g of epoxy-based magnetic microspheres in a three-necked flask, add 100 mL of 0.6 M aminocaproic acid solution with a pH of 9.6, and mechanically stir at room temperature for 8 h to end the reaction. The obtained product was separated and removed by magnetic suction, and washed alternately with deionized water, 1M NaCl, and deionized water several times to obtain carboxyl magnetic microspheres with a magnetic microsphere body-arm chain-carboxyl structure.

[0048] Different arm chains can be prepared by changing the added carboxyl modification reagent, such as changing aminocaproic acid to 2-aminoacetic acid, 3-aminopropionic acid, 4-aminobutyric acid, 5-aminopentanoic acid, 7-aminoheptanoic acid, etc. Long carboxyl magnetic microspheres.

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Abstract

The invention discloses a magnetic microsphere for coupling biological ligand containing primary amino group and a preparation method thereof. The surface of the magnetic microsphere provided by the invention contains NHS (N-hydroxy succinimide) groups; and the magnetic microsphere, without EDC / NHS or glutaraldehyde activation, can directly perform covalent coupling with biological ligand containing primary amino group, and has the advantages of simple operation, mild coupling conditions and fast efficient coupling with biological ligand. In addition, NHS is connected with the microsphere surface by an arm chain with 5-50 Angstrom, so as to reduce steric hindrance during interaction of the biological ligand with an objective bonder and greatly improve the activity and effective utilization rate of the biological ligand. The NHS modified magnetic microsphere has surface NHS content of 50-1000 mumol / g, which is higher compared with the prior art; and the magnetic microsphere can couple with more biological ligand, so as to improve the combination amount and sensitivity of magnetic microsphere with the target substance.

Description

technical field [0001] The invention relates to a magnetic microsphere and a preparation method thereof, in particular to a magnetic microsphere which can be quickly and efficiently coupled with a bioligand containing a primary amino group and a preparation method of the magnetic microsphere, belonging to the field of biological materials. Background technique [0002] Biomagnetic microspheres are a new immunological technology developed in recent years. Its principles include: firstly, using core / shell methods to synthesize magnetic microspheres whose shells are made of inorganic materials, polymer materials, biological macromolecules, etc. These magnetic microspheres It has good physical and chemical stability, biocompatibility and superparamagnetism, and it is easy to use fluorescent substances for post-labeling; secondly, the functional groups on the surface of magnetic microspheres such as amino, carboxyl, epoxy, aldehyde, and mercapto Covalent coupling or physical coat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C01B33/12H01F1/42C08J7/12
Inventor 任辉谭建雄
Owner BEAVERNANO TECH
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