A kind of synthesis method of strong acid agar-based chromatography medium with high adsorption capacity

A synthesis method and high adsorption technology, applied in chemical instruments and methods, ion exchange, microcapsule preparations, etc., can solve the problems of slow flow rate, uneven distribution of active groups and ligands, limited pressure, etc., and achieve the degree of functionalization. High, save the preparation cost, simplify the effect of the preparation process

Active Publication Date: 2020-09-22
AMICOGEN CHINA BIOPHARM CO LTD
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  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

The agarose gel microspheres without any cross-linking can bear limited pressure and the flow rate is slow; the mechanical strength of the cross-linked agarose gel microspheres is obviously enhanced, but because most of the conventional cross-linking agents are oil-soluble, it is difficult to It is difficult to uniformly cross-link in the pores of agarose gel microspheres with a water content of about 95%, so the microsphere structure and surface cross-linking degree are different and affect their mechanical properties; at the same time, a higher degree of cross-linking will limit the activation Diffusion of reagents into the ball leads to uneven distribution of active groups and ligands and a decrease in density, which in turn affects the separation performance of the product

Method used

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  • A kind of synthesis method of strong acid agar-based chromatography medium with high adsorption capacity
  • A kind of synthesis method of strong acid agar-based chromatography medium with high adsorption capacity
  • A kind of synthesis method of strong acid agar-based chromatography medium with high adsorption capacity

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] a) Water phase preparation

[0036] Prepare a 4% agarose aqueous solution (containing 10% KCl), heat to 92° C., the agarose is completely dissolved, add 3-chloro-2-hydroxypropanesulfonate solid and stir until dissolved to obtain an aqueous phase. The mass ratio of the sodium 3-chloro-2-hydroxypropanesulfonate to agarose is 2:1.

[0037] b) Suspension preparation

[0038] The water phase was poured into the toluene oil phase at 70°C, and the volume ratio of the oil phase to the water phase was 8:1 to obtain a suspension; the polyvinyl acetate and oleic acid contained in the toluene had a mass of 2% of the mass of toluene, respectively. % and 0.8%;

[0039] c) Strong acid functionalization and crosslinking reaction

[0040] Add epichlorohydrin to the suspension in step b), and dropwise add a NaOH solution with a mass concentration of 50%, and drop it within 1 hour; keep the temperature at 70° C., and react for 3 hours to obtain a cross-linked reaction product; The mas...

Embodiment 2

[0046] a) Water phase preparation

[0047] Prepare agarose aqueous solution (containing 10% KCl) with a mass concentration of 4%, heat to 95° C., the agarose is completely dissolved, add 3-chloro-2-hydroxypropanesulfonate solid and stir until dissolved to obtain an aqueous phase. The mass ratio of the sodium 3-chloro-2-hydroxypropanesulfonate to agarose is 2.5:1.

[0048] b) Suspension preparation

[0049] Pour the water phase into the toluene oil phase at 70°C, and the volume ratio of the oil phase to the water phase is 7:1 to obtain a suspension; the polyvinyl acetate and oleic acid contained in the toluene have a mass of 100% of the mass of toluene, respectively. 1.5% and 1%;

[0050] c) Strong acid functionalization and crosslinking reaction

[0051] Add epichlorohydrin to the suspension in step b), and dropwise add NaOH solution with a mass concentration of 50%, and dropwise addition is completed within 1 hour; keep the temperature at 70° C., and react for 4 hours to o...

Embodiment 3

[0057] a) Water phase preparation

[0058] Prepare an aqueous solution of agarose (containing 10% KCl) with a mass concentration of 4%, heat it to 93° C., dissolve all the agarose, add 3-chloro-2-hydroxypropanesulfonate solid and stir until dissolved to obtain an aqueous phase; The mass ratio of sodium 3-chloro-2-hydroxypropanesulfonate to agarose is 1.5:1.

[0059] b) Suspension preparation

[0060] Pour the water phase into the toluene oil phase at 70°C, and the volume ratio of the oil phase to the water phase is 6-8:1 to obtain a suspension; the polyvinyl acetate and oleic acid contained in the toluene have a mass of toluene 1.8% and 1.2% of mass;

[0061] c) Strong acid functionalization and crosslinking reaction

[0062] Add epichlorohydrin to the suspension in step b), and dropwise add NaOH solution with a mass concentration of 50%, and dropwise addition is completed within 1 hour; keep the temperature at 70° C., and react for 6 hours to obtain a cross-linked reaction...

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Abstract

The invention relates to the field of biological separation of biological engineering, cells and cell pharmaceutical carriers and particularly relates to a method for synthesizing a high adsorption strong acid agar-based chromatography medium. An agarose solution is used as a water phase. Methylbenzene is an oil phase. Sodium 3-chloro-2-hydroxypropanesulfonate and epichlorohydrin realizes a functionalized and crosslinking reaction for forming agarose microspheres. The strong acid functionalization, a crosslinking reaction and a ball formation process are integrated. Finally, the balls are post-treated so that products are obtained. The method has simple processes and saves the preparation cost. The high adsorption strong acid agar-based chromatography medium has good mechanical strength, uniform distribution of functional groups and a high adsorption amount.

Description

technical field [0001] The invention relates to the field of biological separation of biological engineering, cells and drug carriers thereof, in particular to a method for synthesizing a strong-acid agar-based chromatographic medium with high adsorption capacity. Background technique [0002] In recent years, the upstream cell culture and expression technology has been continuously matured, and the production scale has continued to expand. The rapid development of biotechnology has brought enormous pressure to the downstream separation and purification, requiring the downstream to have greater processing capacity and higher production efficiency. [0003] Ion-exchange chromatography (Ion-exchange chromatography, IEC) uses the ion-exchange medium as the stationary phase, and realizes the separation of solutes according to the difference in the strength of the electrostatic interaction between the charged solute in the mobile phase and the charged group of the medium. Due to ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08J3/24C08L5/12C08K5/1515C08B37/12B01J39/05B01J39/18B01J39/26B01J13/14
CPCB01J13/14B01J13/20B01J39/05B01J39/18B01J39/26C08B37/0039C08J3/24C08J2305/12C08K5/1515
Inventor 刘书霞罗文军王玲杨申永
Owner AMICOGEN CHINA BIOPHARM CO LTD
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