Core-shell magnetic nanoparticle with high selectivity for antibody separation and preparation method of core-shell magnetic nanoparticle

A magnetic nanoparticle, high-selectivity technology, applied in analytical materials, measuring devices, instruments, etc., can solve the problems of low reuse rate, poor chemical stability, and high cost, achieve high selectivity, good binding ability, and improve roughness. degree of effect

Pending Publication Date: 2022-04-08
智享生物(苏州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although protein A chromatography has been regarded as the "gold standard" for antibody capture and purification, it still suffers from low capacity, high cost, low reusability and poor chemical stability.

Method used

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  • Core-shell magnetic nanoparticle with high selectivity for antibody separation and preparation method of core-shell magnetic nanoparticle

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Embodiment 1

[0023] A method for preparing a core-shell magnetic nanoparticle with high selectivity for antibody separation, comprising the following steps:

[0024] (1) Slowly add 1L of a mixed solution containing 0.015mol ferric chloride and 0.005mol ferrous chloride dropwise into 100ml of a concentration of 1.4mol / L and a temperature of 60°C of sodium hydroxide solution, vigorously stir while adding, Stir the reaction for 2 hours after the dropwise addition, cool to room temperature after the reaction, and wash the precipitate to obtain nano-ferric oxide;

[0025] (2) Disperse 1mol of nanometer iron ferric oxide in 500ml of absolute ethanol, then dropwise add 0.8mol of tetraethyl orthosilicate, and dropwise add 10ml of a mass concentration of 30% ammonia solution, stir and react at room temperature for 10h, and the reaction ends Finally, the reaction solution is filtered, and the precipitate is washed with deionized water and absolute ethanol in sequence, and dried to obtain ferric oxid...

Embodiment 2

[0030] A method for preparing a core-shell magnetic nanoparticle with high selectivity for antibody separation, comprising the following steps:

[0031] (1) Slowly add 1L of a mixed solution containing 0.025mol ferric chloride and 0.015mol ferrous chloride dropwise into 100ml of a concentration of 1.6mol / L and a temperature of 70°C in sodium hydroxide solution, vigorously stir while adding, Stir and react for 3 hours after the dropwise addition, cool to room temperature after the reaction, and wash the precipitate to obtain nano-ferric oxide;

[0032] (2) Disperse 1mol of nanometer iron ferric oxide in 500ml of absolute ethanol, then dropwise add 1mol of tetraethyl orthosilicate, and dropwise add 10ml of a mass concentration of 30% ammonia solution, stir and react at room temperature for 15h, after the reaction finishes Filter the reaction solution, wash the precipitate with deionized water and absolute ethanol in sequence, and dry to obtain ferric oxide / silicon oxide nanopart...

Embodiment 3

[0037] A method for preparing a core-shell magnetic nanoparticle with high selectivity for antibody separation, comprising the following steps:

[0038] (1) Slowly add 1L of a mixed solution containing 0.015mol ferric chloride and 0.015mol ferrous chloride dropwise into 100ml of a concentration of 1.4mol / L and a temperature of 70°C in sodium hydroxide solution, vigorously stir while adding, Stir the reaction for 2 hours after the dropwise addition, cool to room temperature after the reaction, and wash the precipitate to obtain nano-ferric oxide;

[0039] (2) Disperse 1mol of nanometer iron ferric oxide in 500ml of absolute ethanol, then dropwise add 1mol of tetraethyl orthosilicate, and dropwise add 10ml of a mass concentration of 30% ammonia solution, stir and react at room temperature for 11h, after the reaction finishes Filter the reaction solution, wash the precipitate with deionized water and absolute ethanol in sequence, and dry to obtain ferric oxide / silicon oxide nanop...

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Abstract

The invention discloses a preparation method of core-shell magnetic nanoparticles with high selectivity for antibody separation, which comprises the following steps: preparing ferroferric oxide / silicon oxide nanoparticles with a core-shell structure, performing amine functionalization treatment on the ferroferric oxide / silicon oxide nanoparticles, and finally coupling IgG2a on the surfaces of the ferroferric oxide / silicon oxide nanoparticles. The preparation method comprises the following steps: dispersing IgG2a coupled magnetic nanoparticles in a PBS (Phosphate Buffer Solution), adding tetraethyl silicate, stirring and reacting, adding a mixed solution of N-propyltriethoxysilane, benzyltriethoxysilane and (3-aminopropyl)-triethoxysilane, reacting, dropwise adding a mixed solution of methanol, PEG-4 and NHS (N-Hydroxysuccinimide) after the reaction is finished, continuously stirring, washing precipitates by adopting the PBS, and drying, thereby obtaining the IgG2a coupled magnetic nanoparticles. And finally, adding the precipitate into a mixed solution of a sodium hydroxide solution and a polyethylene glycol octylphenol ether solution for treatment to prepare magnetic nanoparticles, and re-dispersing the magnetic nanoparticles into a PBS buffer solution for later use. The core-shell magnetic nanoparticle provided by the invention has many surface binding sites and high selectivity, and can effectively purify a monoclonal shell.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a highly selective core-shell magnetic nanoparticle for antibody separation and a preparation method thereof. Background technique [0002] Monoclonal antibody, as a highly specific and effective biological targeted drug, has already accounted for half of the top ten drugs in the world. With the help of the construction of high-expression cell lines, large bioreactors and the application of new culture techniques, the antibody titer in the supernatant of Chinese hamster ovary cell fed-batch culture has been increased by more than 100 times, reaching 3-13g / L, PER The antibody titer in the supernatant of cell perfusion culture can be as high as 27g / L. In sharp contrast to this, protein A chromatography, which is the main means of antibody capture and purification, has an average annual growth rate of antibody binding capacity of only about 5.5%. Antibody capture and purification ref...

Claims

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Application Information

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IPC IPC(8): G01N33/531G01N33/543
Inventor 诸葛鑫
Owner 智享生物(苏州)有限公司
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