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Macroporous agarose microspheres and preparation method thereof

A kind of technology of agarose microsphere, agarose

Active Publication Date: 2013-04-24
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the solid particle porosity method generally has problems such as cumbersome process, many closed pores in the prepared medium, and poor penetration of the pore structure.
Moreover, a large amount of inorganic particle porogen is introduced during the preparation process, and it is difficult to completely remove the microspheres after treatment, which also brings many problems to the later application.

Method used

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  • Macroporous agarose microspheres and preparation method thereof
  • Macroporous agarose microspheres and preparation method thereof
  • Macroporous agarose microspheres and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1: Preparation of 2wt% macroporous agarose microspheres by swelling of micelles

[0064]Accurately weigh 1.0 g of agarose powder, add 50 mL of deionized water, and heat to dissolve to obtain an agarose solution with a concentration of 2 wt%. The temperature of the system was adjusted to 60° C., and 7.5 g of surfactant Tween85 (HLB=11.0) was added thereto so that its volume concentration in the agarose solution was 15%. Under the condition of 100rpm, fully stir for 0.5h, so that the surfactant is uniformly dispersed in the water phase to form micelles. The oil phase was 300 mL of cyclohexane, which contained 4 wt% of the oil phase emulsifier Arlacel83. Pour the above water phase into the oil phase preheated to 55°C in advance, and stir at 300 rpm for 10 min to prepare a W / O emulsion. Then, the emulsion was transferred to a constant temperature water bath shaker, and the reaction was continued for 1 h at 55° C. and 250 rpm, so that the micelles were fully absorb...

Embodiment 2

[0068] Example 2: Preparation of 4wt% macroporous agarose microspheres by swelling of micelles

[0069] Accurately weigh 2.0 g of agarose powder, add 50 mL of deionized water, and heat to dissolve to obtain an agarose solution with a concentration of 4 wt%. The temperature of the system was adjusted to 90° C., and 20.0 g of surfactant Tween40 (HLB=15.6) was added thereto to make its volume concentration in the agarose solution 40%. Under the condition of 150rpm, fully stir for 0.5h, so that the surfactant is uniformly dispersed in the water phase to form micelles. The oil phase is 500mL of liquid paraffin, which contains 2wt% oil phase emulsifier Span80. Pour the above water phase into the oil phase preheated to 60°C in advance, and stir at 800 rpm for 5 min to prepare a W / O emulsion. Then, the emulsion was transferred to a constant temperature water bath shaker, and the reaction was continued for 3 hours at 60° C. and 150 rpm, so that the micelles were fully absorbed by oil...

Embodiment 3

[0072] Example 3: Preparation of 6wt% macroporous agarose microspheres by swelling of micelles

[0073] Accurately weigh 3.0 g of agarose powder, add 50 mL of deionized water, and heat to dissolve to obtain an agarose solution with a concentration of 6 wt%. Dissolve 1.0g SDS in 9.0g Triton X-100 (HLB=17.0) at 80°C, and then add it to the agarose solution at a constant temperature of 80°C, so that the volume concentration of the surfactant phase in the agarose solution is 20%. Under the condition of 200rpm, fully stir for 0.5h, so that the surfactant is uniformly dispersed in the water phase to form micelles. The oil phase was 200 mL of a mixture of cyclohexane and carbon tetrachloride at a volume ratio of 3:1, which contained 1 wt % oil phase emulsifier PO-500. Pour the above water phase into the oil phase preheated to 65°C, and stir at 900 rpm for 5 min to prepare a W / O emulsion. Then, the emulsion was transferred to a constant temperature water bath shaker, and the reacti...

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Abstract

The invention relates to a preparation method and a product of macroporous agarose microspheres. The preparation method is characterized by comprising the following steps: adding a high-concentration surface active agent to an agarose aqueous solution to form a large number of micelles in a water phase; dispersing the water phase into an oil phase; and then preparing the macroporous agarose microspheres according to the characteristic that the micelles of the surface active agent in the water phase can absorb the oil and then swells. The prepared product remains the original mesh shaped gel pores of the agarose gel and also has macropores at hundred-nanometer level (100 to 500nm). The preparation method of the macroporous agarose microspheres, provided by the invention, is simple, and the aperture of the product can be controlled; the prepared macroporous agarose microspheres remain the original excellent performances of the agarose gel and greatly increases the aperture, and are ideal liquid phase chromatogram stationary phase medias and excellent immobilized enzyme carriers; and the macroporous agarose microspheres serving as the liquid phase chromatogram media especially can run at higher flow rate, and can realize quick and efficient separation of biomolecule, in particular biomacromolecule.

Description

technical field [0001] The invention relates to the preparation technology of agarose gel medium, in particular, the invention relates to a macroporous agarose microsphere and a preparation method thereof. Background technique [0002] In recent years, with the rapid development of biotechnology, the upstream cell culture and expression technology has continued to mature, and the production scale has continued to expand, which has brought enormous pressure to the downstream separation and purification, requiring greater processing capacity and higher production downstream. efficiency. Chromatographic separation medium is the core of downstream separation and purification, and its kinetic performance depends on the internal pore structure. [0003] Agarose microspheres have become the most widely used liquid chromatography stationary phase matrix in the field of biochemical separation due to their good biocompatibility and open network structure. However, there are also lim...

Claims

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Application Information

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IPC IPC(8): B01J13/02B01J20/24B01J20/30B01J20/291C08J9/00C08J3/24C08L5/12
Inventor 马光辉赵希吴颉苏志国崔金梅周炜清
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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