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Agar gel microspheres and preparation method thereof

An agar gel and microsphere technology, applied in chemical instruments and methods, other chemical processes, etc., can solve the problems of poor mechanical strength of agar gel bare spheres, high cost of biochemical separation and purification media, and achieve difficult to buy, strong rigidity, cost reduction effect

Active Publication Date: 2014-07-23
BEIJING UNIV OF CHEM TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] One of the purposes of the present invention is to solve the problem of poor mechanical strength of agar gel bare balls, and provide a method of using long and short cross-linking agents for 2 cross-linking, determine the amount and ratio of long and short cross-linking agents, thereby obtaining stronger rigidity, Agar microspheres with more stable structure and better performance
[0005] The second object of the present invention is to solve the problem that the cost of biochemical separation and purification medium is too high, and a kind of medium that utilizes agar gel microspheres to separate and purify biochemical macromolecules and natural active small molecules is provided

Method used

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  • Agar gel microspheres and preparation method thereof
  • Agar gel microspheres and preparation method thereof
  • Agar gel microspheres and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1: secondary cross-linking method and steps

[0048] Take 10g of cross-linked 6B agar gel microspheres as an example:

[0049] Take 6% agar gel microspheres, wash them with water and filter dry, then weigh 10 g of bare agar spheres in a three-necked flask, add water with the same volume as the bare agar spheres, and pour it into the flask, add 2ml of cross-linking agent1,4 -Butanediol diglycidyl ether, mix well. Stir at 60-100rpm for 2-3h, then stand overnight at 20°C. [This process is a process in which the long cross-linking agent diffuses into the interior of the agar microspheres, so that the cross-linking agent can evenly diffuse into the interior of the gel microspheres, making the cross-linking process easier]

[0050] Stir mechanically for 1 hour at room temperature at 20°C; add 0.6ml of NaOH solution with a mass percent concentration of 40%-50% dropwise to the system within 20 minutes and add 0.15g of NaBH 4 ;React for 4 hours, mechanically stir t...

Embodiment 2

[0055] Embodiment 2: flow rate pressure after agar gel microsphere packing column

[0056] The uncrosslinked, primary crosslinked and secondary crosslinked agar gel microsphere packing column (the inner diameter of the column is 1cm, and the column height is 28cm) is connected to the AKTA chromatographic system, and the flow rate increases to each gel. After the highest limit of the gel column, measure the relationship between the flow rate and pressure curve, figure 1 It is the pressure-velocity curve graph after uncrosslinked, primary crosslinked and secondary crosslinked, the square is the bare ball without crosslinking, the circle is after the primary crosslinking, and the triangle is after the secondary crosslinking. The results can be seen To: Compared with the effect after the first cross-linking, the microspheres after the second cross-linking have higher mechanical strength, are more able to withstand pressure and high flow rate, and are more suitable for amplificatio...

Embodiment 3

[0057] Example 3: SEM analysis of agar microspheres after secondary crosslinking

[0058] Depend on Figure 2-4 It can be seen that most of the uncrosslinked bare balls have shriveled after drying and cannot maintain their complete spherical properties. However, most of the uncrosslinked bare balls can maintain their complete spherical shape after drying, but their surface structure has changed. There were different degrees of depressions, but the microspheres after the secondary crosslinking could still maintain their complete spherical properties after drying, and there was basically no damage under the electron microscope observation.

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Abstract

The invention relates to agar gel microspheres and a preparation method, which belongs to the technical field of biochemical separating and purifying medium preparation. The agar gel microspheres take agar raw spheres as a raw material, and can be prepared through twice crosslinking, a long cross-linking agent I is 1,4-butylene-glycol twice-shrunk glyceryl ether; and a short cross-linking agent II is chloropropylene oxide. The preparation method comprises the following steps: washing agar raw spheres and filtering to a dry state, adding the cross-linking agent 1,4-butylene-glycol twice-shrunk glyceryl ether, uniformly mixing; stirring under mechanical stirring and standing at room temperature overnight; adding a NaOH solution drop by drop and adding NaBH4, reacting for 4 hours and performing mechanical stirring, heating a system to the temperature of 39 DEG C, reacting for 4 hours under the temperature; adding 2ml of cross-linking agent chloropropylene oxide in the system drop by drop, heating to the temperature of 43 DEG C, mixing for half hour, mixing while performing mechanical stirring; adding NaOH drop by drop and adding NaBH4 for uniformly mixing, continuously reacting for 12 hours at the temperature of 43 DEG C; continuously adding the NaOH solution drop by drop, adding chloropropylene oxide for mechanical stirring and reacting , and then washing for multitime to neutrality. The agar gel microspheres have the advantages of stronger rigidity, stable structure and better performance.

Description

technical field [0001] The invention belongs to the technical field of biochemical separation and purification medium preparation, and in particular relates to an agar gel bare sphere and a crosslinking method thereof. Background technique [0002] There are many kinds of biochemical separation media, and the earliest biochemical separation media is natural polysaccharide media. This type of medium contains a large amount of hydroxyl groups, which is especially suitable for the separation and purification of macromolecular substances in aqueous solutions. At the same time, as an active group, hydroxyl groups can also undergo many chemical reactions, thereby preparing more types of characteristic media. The polysaccharide medium has good compatibility with biomacromolecules, and can effectively improve the recovery rate under the condition of ensuring the activity of the target product. Existing natural polysaccharide media mainly include dextran, cellulose, agarose, chitosa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08J3/24C08L5/12B01J20/24
Inventor 谭天伟葛春玲吕永琴张帆
Owner BEIJING UNIV OF CHEM TECH
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