Immunomagnetic nano particle cell separator, and its preparing method and use

A magnetic nanoparticle and cell separator technology, applied in biochemical instruments, biochemical equipment and methods, bioreactor/fermenter combination, etc., can solve the problem of inconvenient separation of CD34+ cells, and there is no report on the structure of magnetic nanoparticle cell separator And other issues

Inactive Publication Date: 2006-10-25
SHANGHAI NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

There are many methods for isolating CD34+ cells from umbilical cord blood, but all of them have various disadvantages, and CD34+ cells cannot be isolated conveniently, quickly and efficiently
A

Method used

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  • Immunomagnetic nano particle cell separator, and its preparing method and use
  • Immunomagnetic nano particle cell separator, and its preparing method and use
  • Immunomagnetic nano particle cell separator, and its preparing method and use

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preparation example Construction

[0030] The method for preparing an immunomagnetic nanoparticle cell separator of the present invention includes the following steps:

[0031] (1) Preparation of magnetic nanoparticles

[0032] Separately prepare FeSO with double distilled water 4 .7H 2 O and FeCl 3 .6H 2 O mixed solution and NaOH solution. Fe in the mixed solution of iron salt 2+ The ion concentration is 0.1-0.2mol / l, Fe 3+ The concentration of ions is 0.1-0.3 mol / l, and the concentration of NaOH solution is 2-3 mol / l. Under vigorous stirring, a NaOH solution whose volume is half of the volume of the mixed salt solution is slowly added dropwise to the mixed salt solution. The obtained solid precipitate was aged for 12h at 40℃-60℃, washed with double distilled water several times, filtered and then dried at 40℃-80℃ for 24h, then ground in an agate mortar Get the product.

[0033] (2) Silica in γ-Fe 2 O 3 Surface modification

[0034] TritonX-100, n-hexanol and cyclohexane are uniformly mixed in a ratio of 1:(1-3):(...

Embodiment 1

[0049] Method for preparing core layer of magnetic nano particles

[0050] Using improved chemical co-precipitation to prepare the core layer of magnetic particles, the specific method is as follows: separately prepare FeSO with double distilled water 4 .7H 2 O and FeCl 3 .6H 2 O mixed solution and NaOH solution. Fe in the mixed solution of iron salt 2+ The ion concentration is 0.1-0.2mol / l, Fe 3+ The concentration of ions is 0.1-0.3 mol / l, and the concentration of NaOH solution is 2-3 mol / l. Under vigorous stirring, a NaOH solution whose volume is half of the volume of the mixed salt solution is slowly added dropwise to the mixed salt solution. The obtained solid precipitate was aged for 12 hours at 40°C-60°C, washed with double distilled water several times, filtered and then dried at 40°C-80°C for 24 hours, and then ground in an agate mortar. Get the product.

Embodiment 2

[0052] Preparation method of core-shell type nucleic acid magnetic nano particles

[0053] TritonX-100, n-hexanol and cyclohexane are uniformly mixed in a ratio of 1:2:5 to form a transparent and stable microemulsion system. Put the above microemulsion system in ultrasonic for 30-60 minutes, and then add 0.5g of γ-Fe to it 2 O 3 , After ultrasonic treatment for 6 minutes, take out the upper layer and pour it into a three-necked flask, stir for 30 minutes to make it uniform. Dilute 1ml of a certain concentration of concentrated ammonia water with 2ml of double distilled water, slowly add it to the continuously stirred microemulsion after 30 minutes, and continue to stir for 30 minutes to make the ammonia water evenly dispersed in the microemulsion. After 1 hour, 1-3 ml of ethyl orthosilicate was added dropwise to the microemulsion while stirring continuously for 10 hours, and the temperature of the system was kept between 15-30°C. Add acetone to the system to precipitate the partic...

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Abstract

The invention relates to an immunity magnetic nanometer particle cell separator that contains inner nuclear layer, N-(2- amino group ethyl)-3-amino group propyl tri-methoxyl group silicane intermediate layer, and antibody and/or ligand outer shell. The invention also relates the method to use the separator.

Description

Technical field [0001] The present invention relates to a nano biological separator, more specifically, the present invention relates to an immunomagnetic nano particle cell separator. The invention also relates to the preparation method and application of the separator. Background technique [0002] As we all know, the field of nanotechnology is the hottest research field of science and technology today. The application of nanotechnology in the field of biological sciences forms an emerging science and technology-nanobiotechnology, which uses nanotechnology to research and solve major problems in the field of life sciences. Science, it is becoming one of the most important frontier scientific research fields. [0003] Now, the application prospects of magnetic nanoparticles in the field of biotechnology have attracted increasing attention. Magnetic nanoparticles are often used as separation media for magnetic field separation of biological samples. The s...

Claims

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Application Information

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IPC IPC(8): C12N5/00C12M1/00G01N1/02G01N33/50G01N33/53
Inventor 沈鹤柏李兴玉陈伟忻茜
Owner SHANGHAI NORMAL UNIVERSITY
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