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Preparation method of monodisperse agarose superparamagnetic micro-sphere

A superparamagnetic and paramagnetic technology, applied in the field of monodisperse agarose superparamagnetic microsphere preparation, can solve the problems of long incubation time, low purity, lengthy operation steps, etc. control effect

Inactive Publication Date: 2018-07-06
SUZHOU WEIDU BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for scientific researchers, there are many problems in the process of operating agarose microspheres to separate and purify biomolecules: 1) the purity is low, and the porous structure of agarose microspheres is easy to capture various biomolecules; 2) long incubation and binding time , low efficiency; 3) The product yield is low, because the long incubation time and a large number of washing processes will lead to the degradation and inactivation of the target purified product; 4) The operation steps are lengthy and cumbersome; 5) The automatic operation cannot be realized, and the result is vulnerable to Human factors
From the above preparation methods reported in existing patents, it is not difficult to find that there is no agarose magnetic microsphere prepared by the existing method that can meet the requirements of ideal microspheres, which also limits the agarose magnetic microspheres. Balls are widely used in medical treatment and biological separation and purification

Method used

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  • Preparation method of monodisperse agarose superparamagnetic micro-sphere

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Experimental program
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Effect test

Embodiment 1

[0027] 1) Preparation of Fe 3 o 4 Preparation of magnetic nanoparticles and their surface modification:

[0028] 126g FeCl 2 .4H 2 O, 310g FeCl 3 .6H 2 O was dissolved in 1000g of pure water. Preheat the reaction kettle in a constant temperature water bath at 70°C, replace the air in the reaction kettle with nitrogen, add iron salt solution and stir for 30 minutes, then quickly add 500ml ammonia water for precipitation reaction, after the reaction is carried out for 30 minutes, add 0.1wt% polyethylene Pyrrolidone, after 1hr of surface chemical modification, was heated to 80°C and the reaction system was opened to completely evaporate excess ammonia water. After washing with alcohol and water for many times, it was filtered and concentrated into 30wt% aqueous magnetic fluid for later use. The primary particle size of magnetic nanoparticles is 13-20nm, and they are superparamagnetic.

[0029] 2) Preparation of core-shell magnetic nanoparticles by sol-gel method:

[0030]...

Embodiment 2

[0036] 1) Preparation of Fe 3 o 4 Preparation of magnetic nanoparticles and their surface modification.

[0037] 126g FeCl 2 , 310g FeCl 3 Dissolve in 1000g pure water. Preheat the reaction kettle in a constant temperature water bath at 60°C, replace the air in the reaction kettle with nitrogen, add the iron salt solution and stir for 30 minutes, then quickly add 500ml of ammonia water for precipitation reaction, after the reaction is carried out for 30 minutes, add 2wt% sodium alginate , after 0.5hr of surface chemical modification, the temperature was raised to 65°C and the reaction system was opened to completely evaporate the excess ammonia water. After washing with alcohol and water for several times, it was filtered and concentrated into 50wt% aqueous magnetic fluid for later use. The primary particle size of magnetic nanoparticles is 16-25nm, and they are superparamagnetic.

[0038] 2) Preparation of core-shell magnetic nanoparticles by sol-gel method

[0039] Tak...

Embodiment 3

[0045] 1) Preparation of Fe 3 o 4 Preparation of magnetic nanoparticles and their surface modification.

[0046] 126g FeCl 2 .4H 2 O, 310g FeCl 3 .6H 2 O was dissolved in 1000g of pure water. Preheat the reaction kettle in a constant temperature water bath at 80°C, replace the air in the reaction kettle with nitrogen, add iron salt solution and stir for 30 minutes, then quickly add 500ml of ammonia water for precipitation reaction, after the reaction is carried out for 30 minutes, add 1wt% sodium citrate , after 2 hours of surface chemical modification, the temperature was raised to 90°C and the reaction system was opened to completely evaporate the excess ammonia water. After washing with alcohol and water for several times, it was filtered and concentrated into 10wt% aqueous magnetic fluid for later use. The primary particle size of magnetic nanoparticles is 5-10nm, and they are superparamagnetic.

[0047] 2) Preparation of core-shell magnetic nanoparticles by sol-gel...

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Abstract

The invention provides a monodisperse agarose superparamagnetic micro-sphere and a preparation method thereof. The preparation method comprises the following steps: 1) preparing magnetic nano-particles by adopting a co-precipitation method, and performing surface modification to obtain monodisperse water-based magnetic fluid; 2) coating the surfaces of the magnetic nano-particles with an inorganicshell layer with certain thickness by adopting a gel-sol method so as to enhance the acid resistance, alkali resistance and oxidation resistance of the magnetic nano-particles; 3) dispersing the magnetic nano-particles with a core-shell structure in an agarose aqueous solution with certain concentration, extruding the magnetic nano-particles into an oil phase by the high-pressure shear action ofa template method, changing the pore sizes of micro-pores of a template, the extrusion pressure, the flow rate of the oil phase, the variety and concentration of emulsifying agents and other conditions so as to obtain a W / O type emulsion droplet solution with a uniform size, and after performing cooling, washing and vacuum drying to obtain an agarose superparamagnetic micro-sphere cluster; and 4)transferring the micro-sphere cluster into a conical flask, and performing solidification under the action of alkali, a reducing agent and a cross-linking agent to obtain the agarose superparamagneticmicro-sphere.

Description

technical field [0001] The invention shows a method for preparing monodisperse agarose superparamagnetic microspheres applied in the fields of nucleic acid purification and protein separation. Background technique [0002] As a natural polysaccharide chromatographic medium, agarose microspheres have the characteristics of high hydrophilicity, porosity, a large number of activated hydroxyl groups, and low non-specific adsorption. They are widely used in affinity chromatography columns, involving the separation and purification of proteins , nucleic acid, polypeptide, sugar and other water-soluble biomolecules. However, for scientific researchers, there are many problems in the process of operating agarose microspheres to separate and purify biomolecules: 1) the purity is low, and the porous structure of agarose microspheres is easy to capture various biomolecules; 2) long incubation and binding time , low efficiency; 3) The product yield is low, because the long incubation t...

Claims

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Application Information

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IPC IPC(8): C08L5/12C08K9/10C08K3/22B01J13/14
CPCB01J13/14C08K3/22C08K9/10C08K2003/2275C08K2201/01C08K2201/011C08L5/12
Inventor 宋孟杰杜德状
Owner SUZHOU WEIDU BIOTECH CO LTD
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