Size-uniform agarose gel microball and its preparing method

A technology of agarose gel and agarose, which is applied in the field of biochemical separation of bioengineering, can solve the problems of inhomogeneous particle size of agarose gel microspheres, large pressure changes, and poor separation effect, so as to avoid recognition and immune rejection , The pressure is stable and uniform, and the effect of column packing is uniform

Active Publication Date: 2005-07-20
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] This technology solves the problem of poor separation effect caused by the non-uniform particle size of agaros...

Method used

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  • Size-uniform agarose gel microball and its preparing method
  • Size-uniform agarose gel microball and its preparing method
  • Size-uniform agarose gel microball and its preparing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037]The hydrophobic membrane with a pore size of 5.7 μm is immersed in a lipophilic substance to fully wet the porous membrane to ensure that the hydrophobic chains on the membrane are completely stretched. Accurately weigh a certain amount of agarose and a certain amount of NaCl and add it to a certain amount of water, so that the concentration of the agarose is 4%, and the concentration of NaCl is 0.9%, and it is fully dissolved under heating conditions, and the solution is kept constant. The temperature is reserved. Add the oil-soluble emulsifier into 200ml of liquid paraffin, stir until completely dissolved and heat to 50°C as the oil phase. Quickly transfer about 6.0g of the water phase into the membrane emulsification device that has been preheated to 50°C in the oil phase while it is hot, and press it into the oil phase through a hydrophobic microporous membrane with uniform pore size under constant pressure to obtain W / O type emulsion; after the emulsification is co...

Embodiment 2

[0041] The hydrophobic membranes with pore diameters of 4.7 μm, 5.7 μm, 13 μm, and 19.6 μm were soaked in lipophilic substances, so that the porous membranes were fully wet to ensure that the hydrophobic chains on the membranes were completely stretched. Accurately weigh a certain amount of agarose and a certain amount of NaCl and add it to a certain amount of water, so that the concentration of the agarose is 4%, and the concentration of NaCl is 0.9%, and it is fully dissolved under heating conditions, and the solution is kept constant. The temperature is reserved. Add the oil-soluble emulsifier into 200ml of liquid paraffin, stir until completely dissolved and heat to 50°C as the oil phase. Quickly transfer about 6.0g of the water phase into the membrane emulsification device that has been preheated to 50°C in the oil phase while it is hot, and press it into the oil phase through a hydrophobic microporous membrane with uniform pore size under constant pressure respectively. ...

Embodiment 3

[0043] The hydrophobic membrane with a pore size of 4.7 μm is immersed in a lipophilic substance to fully wet the porous membrane to ensure that the hydrophobic chains on the membrane are completely stretched. Accurately weigh a certain amount of agarose and a certain amount of NaCl and add them to a certain amount of water so that the concentrations of the agarose are 2%, 4%, 6% and 8%, respectively, and the concentration of NaCl is 0.9%. Make it fully dissolved, and keep the solution at a certain temperature for later use. Add the oil-soluble emulsifier into 200ml of liquid paraffin, stir until completely dissolved and heat to a certain temperature as the oil phase. Quickly transfer about 6.0g of the water phase into the membrane emulsification device that has been preheated to the required temperature in the oil phase, and press it into the oil phase through a hydrophobic microporous membrane with uniform pore size under constant pressure , to obtain a W / O type emulsion; a...

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Abstract

The present invention provides an agarose gel separation medium which is uniform in size, controllable and has high hydrophility and high porousness, and can be used for making separation and purification of biological active material. Its preparation method is characterized by that it adopts the following steps: (1) dissolving a certain quantity of agarose in a certain quantity of water under the heating condition to obtain agarose aqueous solution with a certain concentration, using pressure to make said aqueous phase by passed through microporous membrane and make it feed into oil phase to obtain emulsion drop with uniform size; and (2). transferringsaid emulsion into cooling device, under the condition of adopting programmed temperature-falling and slow-mechanically stirring process cooling to below 15 deg.C and making gel be solidified so as to obtain the invented agarose gel microspheres with uniform size.

Description

technical field [0001] The invention belongs to the field of biochemical separation of bioengineering Background technique [0002] Agarose gel is a natural polysaccharide chromatography medium, which appeared as early as the 1960s. It has many characteristics of an ideal medium: high hydrophilicity, high porosity, containing more activatable hydroxyl groups, and does not mix with Non-specific adsorption of biomacromolecules is by far the most widely used chromatographic medium [1] . Because agarose contains more activatable hydroxyl groups, it can be connected with different ligands under certain conditions, as a medium for affinity chromatography, hydrophobicity and ion exchange chromatography. With the 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 and efficient chromatographic separation. The separation objects involve wa...

Claims

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Application Information

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IPC IPC(8): B01J13/08
Inventor 马光辉苏志国王连艳周青竹
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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