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

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...

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.

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