Nickel magnetic microsphere and preparation method and application thereof

A technology of magnetic microspheres and microspheres, applied in peptide preparation methods, chemical instruments and methods, magnetic materials, etc., can solve problems such as difficult to wash off, long incubation time, slow contact with target proteins, etc., and achieve strong force Effect

Active Publication Date: 2018-08-17
湖北新纵科病毒疾病工程技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when purifying a large amount of protein, this porous structure will also bring some problems, such as: 1) Antibodies are trapped in the pores and are difficult to wash off, so a lot of washing is required to reduce the background; 2) Washing of immunoprecipitation It is carried out in a microcentrifuge tube, and the exchange between liquids is done by a pipette gun, which is easy to lose samples
3) The contact between the antibody on the microsphere and the target protein is slow, requiring a long incubation time
4) Long incubation time and extensive washing lead to mechanical and biological (protease hydrolysis) loss of the target protein
However, it mainly loads a certain amount of magnetic substances on the basis of traditional agarose microspheres, which is cumbersome to prepare, and the structure and performance of the microspheres are not uniform.

Method used

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  • Nickel magnetic microsphere and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] A kind of nickelized magnetic microsphere, its preparation comprises the steps:

[0027] 1), weigh 1gFe 3 o 4 Particles, add 10mL 0.01mol / L hydrochloric acid solution, activate under ultrasonic conditions for 20min (ultrasonic dispersion power 100W), after magnetic separation, Fe 3 o 4 The particles were washed to neutrality, and then converted to hydrochloric acid-activated Fe 3 o 4 Add 80mL of absolute ethanol, 20mL of ultrapure water, 0.4mL of 25wt% ammonia water and 0.5mL of ethyl orthosilicate to the granules in sequence, react with ultrasound for 2 hours, and wash with ultrapure water until neutral after magnetic separation to obtain magnetic silica microspheres;

[0028] 2) Weigh 1g of the magnetic silica microspheres prepared in step 1), add 80mL of absolute ethanol, 20mL of ultrapure water, 0.4mL of ammonia water and 0.5mL of KH-560 for ultrasonic reaction for 2 hours, and use ultrasonic Washing with pure water to neutrality to obtain epoxidized magnetic s...

Embodiment 2

[0032] 1) Weigh 1g α-Fe 2 o 3 Particles, add 30mL 0.01mol / L hydrochloric acid solution, activate under ultrasonic conditions for 30min (ultrasonic dispersion power 300W), after magnetic separation, Fe 3 o 4 The particles were washed to neutrality, and then converted to hydrochloric acid-activated Fe 3 o 4 Add 130mL of absolute ethanol, 50mL of ultrapure water, 0.7mL of 25wt% ammonia water and 2mL of ethyl orthosilicate to the granules in sequence, react with ultrasound for 2.5h, and wash with ultrapure water until neutral after magnetic separation to obtain magnetic silica microspheres;

[0033] 2) Weigh 1g of the magnetic silica microspheres prepared in step 1), add 130mL of absolute ethanol, 60mL of ultrapure water, 1.1mL of 25wt% ammonia water and 0.9mL of silane coupling agent KH-560 for ultrasonic reaction for 3 hours, After magnetic separation, wash with ultrapure water until neutral to obtain epoxidized magnetic silica microspheres;

[0034] 3), weigh 3g of iminodi...

Embodiment 3

[0037] 1), weigh 1gMgFe 2 o 4Granules, add 40mL 0.01mol / L hydrochloric acid solution, activate under ultrasonic conditions for 40min (ultrasonic dispersion power 500W), after magnetic separation, Fe 3 o 4 Wash the particles until they are neutral, then add 160mL of absolute ethanol, 80mL of ultrapure water, 1.6mL of 25wt% ammonia water and 4mL of tetraethyl orthosilicate to the magnetic substrate activated by hydrochloric acid in sequence, react with ultrasonic for 5h, and use ultrapure water after magnetic separation Washing to neutrality to obtain magnetic silica microspheres;

[0038] 2) Weigh 1g of the magnetic silica microspheres prepared in step 1), add 160mL of absolute ethanol, 80mL of ultrapure water, 1.6mL of 25wt% ammonia water and 4mL of silane coupling agent KH-560 for ultrasonic reaction for 5 hours, magnetic After separation, wash with ultrapure water until neutral to obtain epoxidized magnetic silica microspheres;

[0039] 3) Weigh 4g of iminodiacetic acid ...

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Abstract

The invention relates to a nickel magnetic microsphere and a preparation method and an application thereof. A magnetic silicon dioxide microsphere is prepared by adopting a sol-gel method, wherein themagnetic silicon dioxide microsphere takes magnetic substrate particles as a core, takes tetraethoxysilane as a silicon source and forms a shell through a reaction; the microsphere, used as a template, is modified by epoxy silane coupling agent KH-560 and iminodiacetic acid to form a ligand which is used as a complexing agent for chelating metal nickel ions to prepare the magnetic silicon dioxidemicrosphere with chelated nickel ions on the surface. By adopting the iminodiacetic acid modified silicon dioxide microsphere, multiple coordination sites can be provided for metal nickel ions, so that relatively high acting force between the modified microspheres and the metal nickel ion can be ensured, and the unsaturated coordination sites of metal ions can be acted with protein, thereby realizing that the magnetic silicon dioxide microsphere with chelated nickel ions on the surface can separate protein from a solution.

Description

technical field [0001] The invention relates to a magnetic microsphere, in particular to a nickelized magnetic microsphere, a preparation method and an application thereof. Background technique [0002] Currently, the commercially available porous agarose microspheres with a certain degree of cross-linking are mainly used for the purification of His-tagged proteins, which have a high surface area, allow proteins to contact each other, and can accommodate a large amount of liquid. However, when purifying a large amount of protein, this porous structure will also bring some problems, such as: 1) Antibodies are trapped in the pores and are difficult to wash off, so a lot of washing is required to reduce the background; 2) Washing of immunoprecipitation It is carried out in microcentrifuge tubes, and the exchange between liquids is done through a pipette gun, which is easy to lose samples. 3) The contact between the antibody on the microsphere and the target protein is slow, re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): H01F1/11H01F1/09H01F41/00C07K1/22
CPCC07K1/22H01F1/09H01F1/112H01F41/00
Inventor 王华林张涛张科登
Owner 湖北新纵科病毒疾病工程技术有限公司
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