Preparation method of magnetic sepharose gel microspheres

A technology of agarose gel and agarose, which is applied in the field of preparation of agarose gel particles, can solve problems such as unsatisfactory magnetic responsiveness, and achieve good application prospects, low cost, and narrow distribution range

Active Publication Date: 2013-04-03
无锡百运纳米科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For a long time, people have used ferroferric oxide as the magnetic core to prepare agarose magnetic microspheres, but when the particle size is small, its magnetic responsiveness is not ideal, and the targeting performance needs to be further improved in practical applications.

Method used

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  • Preparation method of magnetic sepharose gel microspheres

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Preparation of active agent-dispersed iron powder: Accurately weigh 0.015mol sodium citrate (4.41g) in 30ml distilled water to make a solution with a concentration of 0.5M, add 0.01mol iron powder (0.56g) to it, and the average iron powder The particle size is 40nm; heat in a water bath at 80°C and stir vigorously for 2 hours, and the stirring speed is 500rpm / min. The magnetic particles were collected by a magnet, washed five times with distilled water and ethanol, and dried in vacuum at 60°C for 6 hours.

[0024] Preparation of magnetic agarose gel microspheres: Dissolve 3.6g span80 in 90mL liquid paraffin, preheat to 70°C in a 250mL capacity three-necked flask; accurately weigh 0.45g agarose and 45mg active agent dispersed iron powder, add 90mL double-distilled water to make 0.5% agarose solution, repeat ultrasonication and microwave heating to dissolve; transfer the water phase to the oil phase, and mechanically stir at 80°C for 0.5h at a stirring speed of 500rpm / min...

Embodiment 2

[0026] Preparation of active agent-dispersed iron powder: Accurately weigh 0.07mol sodium citrate (20.59g) in 260ml distilled water to make a solution with a concentration of 0.27M, add 0.01mol iron powder (0.56g) to it, and the average iron powder The particle size is 50nm; heated in a water bath at 70°C and stirred vigorously for 4 hours, and the stirring speed is 1500rpm / min. The magnetic particles were collected by a magnet, washed five times with distilled water and ethanol, and dried in vacuum at 80°C for 6 hours.

[0027] Preparation of magnetic agarose gel microspheres: Dissolve 3.6g span80 in 337.5mL liquid paraffin, preheat to 80°C in a 2000mL capacity three-necked flask; accurately weigh 0.9g agarose and 180mg active agent dispersed Iron powder, add 22.5mL double-distilled water to make a 4% agarose solution, repeat ultrasonication and microwave heating to dissolve; transfer the water phase to the organic phase, and mechanically stir at 80°C for 1h at a stirring spe...

Embodiment 3

[0030] Preparation of active agent dispersed iron powder: Accurately weigh 0.03mol sodium lauryl sulfate (8.64g) in 120ml distilled water to make a solution with a concentration of 0.25M, add 0.01mol iron powder (0.56g) to it, The average particle size of the iron powder is 2 μm; it is heated in a water bath at 90°C and stirred vigorously for 1 hour, and the stirring speed is 2000 rpm / min. The magnetic particles were collected by a magnet, washed five times with distilled water and ethanol, and dried in vacuum at 40°C for 12 hours.

[0031] Preparation of magnetic agarose gel microspheres: Dissolve 2g span80 in 625mL liquid paraffin, preheat to 80°C in a 1000mL capacity three-neck flask; accurately weigh 1g agarose and 0.5g active agent dispersed iron powder , add 12.5mL double distilled water to make 8% agarose solution, repeat ultrasonication and microwave heating to dissolve; transfer the aqueous phase to the organic phase, and mechanically stir at 90°C for 1h at a stirring...

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Abstract

The invention discloses a preparation method of magnetic sepharose gel microspheres. The preparation method comprises the following steps: (1) preparing ferrous powder with scattered active agent: adding the ferrous powder into surface active agent water solution, heating and mechanically stirring; washing and drying the obtained product to obtain the ferrous powder with scattered active agent; and (2) preparing the magnetic sepharose gel microspheres: dissolving an emulsifying agent span80 into liquid paraffin for preheating; taking sepharose gel and the ferrous powder with scattered active agent, adding secondary distilled water to prepare sepharose solution, conducting ultrasonic scattering and heating and dissolving; adding the dissolved sepharose solution into span80-containing liquid paraffin solution; after the reaction is finished, cooling a sample and conducting slow and mechanical stirring; separating the sample, and washing to obtain the magnetic sepharose gel microspheres. By adopting a reversed phase suspension embedding method, the prepared magnetic sepharose gel microspheres are strong in magnetic responsibility, and have a good effect on the separation and purification of protein.

Description

technical field [0001] The invention relates to the technical field of biomacromolecule separation and purification media, in particular to a method for preparing agarose gel particles wrapped with magnetic microbeads. Background technique [0002] Agarose gel is a natural polysaccharide chromatography medium, which has many characteristics of an ideal medium: high hydrophilicity, high porosity, more activatable hydroxyl groups, and no non-specific adsorption with biological macromolecules. One of the most widely used chromatography media. Since agarose contains more activatable hydroxyl groups, different ligands can be connected under certain conditions, and it can be used as a medium for affinity chromatography, hydrophobicity and ion exchange chromatography. With the continuous deepening of research and application, agarose gel is not only used as a medium for normal pressure and low pressure chromatography, but also can be operated under medium pressure and achieve fast...

Claims

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Application Information

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IPC IPC(8): B01J13/02B01J20/24B01J20/30B01J20/28H01F1/42
Inventor 徐卫宋芳
Owner 无锡百运纳米科技有限公司
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