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Preparation method and storage method of immunomagnetic beads for leucocyte sorting

A technology of immunomagnetic beads and white blood cells, which is applied in cell dissociation methods, biochemical equipment and methods, animal cells, etc., can solve the problems of poor stability and specificity, and achieve good stability, good specificity, and dispersion sexually superior effect

Inactive Publication Date: 2017-12-05
安徽安龙基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The technical problem to be solved by the present invention is to provide a method for preparing and storing immunomagnetic beads for sorting white blood cells in order to overcome the defects of poor stability and poor specificity in the prior art

Method used

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  • Preparation method and storage method of immunomagnetic beads for leucocyte sorting

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preparation example Construction

[0050] The invention discloses a method for preparing immunomagnetic beads for sorting leukocytes. The specific steps are as follows:

[0051] 1. Synthesis of oleic acid-Fe 3 o 4

[0052] (1.1), under the condition of nitrogen, FeCl 2 4H 2 O, FeCl 3 ·6H 2 O was dissolved in 800ml of distilled water, heated to reflux, and when heated to 90°C, while mechanically stirring, 346ml of NaOH solution was added to the above solution;

[0053] (1.2), add 20ml oleic acid, under the condition of 90 ℃, stop the reaction after stirring for 1 hour;

[0054] (1.3) After the solution is cooled to room temperature, apply an external magnetic field to absorb the precipitate, and then wash it with distilled water for 1-7 times, and the product is oleic acid-Fe 3 o 4 ;

[0055] 2. Fe 3 o 4 / PVAc magnetic microsphere preparation

[0056] (2.1), add 100ml of water to the four-necked flask equipped with agitator, condenser, thermometer and air guide tube, heat and reflux in a water bath, ...

Embodiment 1

[0085] The invention discloses a method for preparing immunomagnetic beads for sorting leukocytes. The specific steps are as follows:

[0086] 1. Synthesis of oleic acid-Fe 3 o 4

[0087] (1.1), under the condition of logical nitrogen, 8.6g FeCl 2 4H 2 O, 23.5g FeCl 3 ·6H 2 O was dissolved in 800ml of distilled water, heated to reflux, and when heated to 90°C, while mechanically stirring, 346ml of NaOH solution was added to the above solution;

[0088] (1.2), add 20ml oleic acid, under the condition of 90 ℃, stop the reaction after stirring for 1 hour;

[0089] (1.3) After the solution is cooled to room temperature, an external magnetic field is applied to absorb the precipitate, and then washed with distilled water for 3 times, the product is oleic acid-Fe 3 o 4 ;

[0090] 2. Fe 3 o 4 / PVAc magnetic microsphere preparation

[0091] (2.1), add 100ml of water to the four-necked flask equipped with agitator, condenser, thermometer and air guide tube, heat and reflux ...

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Abstract

The invention discloses a preparation method and a storage method of immunomagnetic beads for leucocyte sorting. The preparation method comprises the following specific steps: synthesizing oleic acid-Fe3O4; preparing Fe3O4 / PVAc magnetic beads; preparing (Fe3O4 / PVA) / SiO2 magnetic beads; dropwise adding 0.4 ml of 3-aminopropyl triethoxysilane into an ethanol suspension of magnetic beads (Fe3O4 / PVA) / SiO2, stirring at room temperature for 7 h for amination, separating from a reaction medium by the utilization of an applied magnetic field, and cleaning with an ethanol solution for five times for later use; adding aminated magnetic nano-particles into 50 ml of 3% glutaraldehyde PBS solution, stirring at 37 DEG C for 3 h, modifying the surface of the magnetic particles with aldehyde function group through Schiff base formed between amino group and aldehyde group, separating particles from the reaction medium by the utilization of an applied magnetic field after the reaction, repeatedly washing a product with a PBS solution, dispersing the particles in the PBS solution until final concentration is 8 microgram / microliter; carrying out hydrazino-modification of antibody; and preparing the immunomagnetic beads. The storage method is to store the immunomagnetic beads at 4 DEG C. The invention has advantages of good stability, strong specificity and the like.

Description

technical field [0001] The invention relates to the field of preparation of immunomagnetic beads, in particular to a method for preparing and storing immunomagnetic beads for sorting leukocytes. Background technique [0002] Immunomagnetic beads (IMB) technology is a new immunological technology researched at home and abroad in recent years. IMB is a kind of immune microspheres, which are spherical magnetic particles coated with monoclonal antibodies. Binding to the target substance makes it magnetically responsive. This technology was first prepared by John Ugelstar in 1979, and has since been widely used in many fields of medicine, health, scientific research and other fields, and thus triggered a revolution in bioseparation technology. [0003] Because the diameter of magnetic particles is at the nanometer level, they are also called magnetic nanoparticles. In recent years, people have coated the surface of nano magnetic beads with antibodies, which can bind to specific ...

Claims

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Application Information

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IPC IPC(8): C12N5/078G01N33/543
CPCC12N5/0634C12N2509/00G01N33/54326
Inventor 韦玉军李航陆宝石苏军吴远航
Owner 安徽安龙基因科技有限公司
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