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Compound immune magnetic beads for separation and purification of hepatic stellate cells and preparation method of compound immune magnetic beads

A technology of hepatic stellate cells and immunomagnetic beads, which is applied in the field of composite immunomagnetic beads and their preparation, can solve the problems of poor antibody stability, low antibody grafting rate, easy agglomeration, etc., achieve good suspension and dispersion, and improve dispersion performance. , the effect of reducing weight

Active Publication Date: 2018-06-29
ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current immunomagnetic beads have poor dispersion performance and are prone to agglomeration. The antibody grafting rate on the immunomagnetic beads is low, and the stability of binding to antibodies is poor.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Fe 3 o 4 Preparation of microspheres: Take 0.1moL Fe 2 (SO 4 ) 3 and 0.1moL FeSO 4 ·7H 2 O mix evenly, add 500mL of distilled water and stir until completely dissolved to obtain a mixed solution, take 100g of egg white and stir into a milky white uniform state, stir and add to the mixed solution drop by drop, stir and react at 60-65°C for 30 hours, and dry under reduced pressure , ground, and transferred to a muffle furnace at 450°C for 6 hours, then took it out and cooled it with ice-water mixture, washed it twice with distilled water and absolute ethanol, and put the dried powder into the reactor. Under the conditions of flow rate 80sccm, pressure 3Pa, and power 80W, conduct plasma treatment for 10min to obtain Fe 3 o 4 Microspheres.

[0030] The preparation of carboxylated graphene is as follows: get graphene and add concentration and be that in the sodium hydroxide solution of 0.3mol / L ultrasonic dispersion 3h, then add chloroacetic acid ultrasonic reaction ...

Embodiment 2

[0036] Fe 3 o 4 Preparation of microspheres: Take 0.1moL Fe 2 (SO 4 ) 3 and 0.1moL FeSO 4 ·7H 2 O mix evenly, add 500mL of distilled water and stir until completely dissolved to obtain a mixed solution, take 100g of egg white and stir into a milky white uniform state, stir and add to the mixed solution drop by drop, stir and react at 60-65°C for 30 hours, and dry under reduced pressure , ground, and transferred to a muffle furnace at 450°C for 6 hours, then took it out and cooled it with ice-water mixture, washed it twice with distilled water and absolute ethanol, and put the dried powder into the reactor. Under the conditions of flow rate 120sccm, pressure 8Pa, and power 100W, conduct plasma treatment for 15min to obtain Fe 3 o 4 Microspheres.

[0037] The preparation of carboxylated graphene is the same as embodiment one.

[0038] Graphene / Fe 3 o 4 Preparation of microspheres: Take 1.5g Fe 3 o 4 The microspheres and 2.0g carboxylated graphene were stirred and di...

Embodiment 3

[0043] Fe 3 o 4 Preparation of microspheres: Take 0.1moL Fe 2 (SO 4 ) 3 and 0.1moL FeSO 4 ·7H 2 O mix evenly, add 500mL of distilled water and stir until completely dissolved to obtain a mixed solution, take 100g of egg white and stir into a milky white uniform state, stir and add to the mixed solution drop by drop, stir and react at 60-65°C for 30 hours, and dry under reduced pressure , ground, and transferred to a muffle furnace at 450°C for 6 hours, then took it out and cooled it with ice-water mixture, washed it twice with distilled water and absolute ethanol, and put the dried powder into the reactor. Under the conditions of flow rate 100sccm, pressure 5Pa, and power 90W, conduct plasma treatment for 13min to obtain Fe 3 o 4 Microspheres.

[0044] The preparation of carboxylated graphene is the same as embodiment one.

[0045] Graphene / Fe 3 o 4 Preparation of microspheres: Take 2.0g Fe 3 o 4 The microspheres and 3.5g carboxylated graphene were stirred and dis...

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Abstract

The invention relates to the technical field of cell separation and purification materials and particularly relates to compound immune magnetic beads for separation and purification of hepatic stellate cells and a preparation method of the compound immune magnetic beads. The compound immune magnetic beads are prepared by grafting antibodies on magnetic bead granules of which the granule sizes are50-1500nm, wherein the antibodies include CD11b and CD146 antibodies, and the inner cores of the magnetic bead granules are Fe3O4 micro-spheres including carboxylated graphene layers, modified chitosan layers and sodium alginate layers from inside to outside. The antibodies are grafted on the magnetic bead granules to obtain the compound immune magnetic beads; the magnetic bead granules contain more carboxyl functional groups, have good dispersion properties in solutions and can not aggregate easily; the antibodies have a higher grafting ratio on the magnetic bead granules and have higher binding stability; and the compound immune magnetic beads can be used for separation and purification of hepatic stellate cells so as to obtain hepatic stellate cells with higher purity and better activity.

Description

technical field [0001] The invention relates to the technical field of cell separation and purification materials, in particular to composite immunomagnetic beads for separation and purification of hepatic stellate cells and a preparation method thereof. Background technique [0002] Liver cells are divided into hepatic parenchymal cells and non-parenchymal cells, including hepatic sinusoidal endothelial cells (LSECs), Kupffer cells (KCs), hepatic stellate cells (HSCs) and crypt cells. The shape of hepatic stellate cells is irregular, the cell body is round or irregular, and several stellate processes often protrude to surround the hepatic sinusoids. There are 1 to 14 lipid droplets with a diameter of about 1.0 to 2.0 μm in the hepatic stellate cytoplasm. The lipid droplets are rich in vitamin A and can spontaneously fluoresce blue-green under ultraviolet excitation. [0003] Since the Anderson invasive shearing method was used to separate hepatocytes from the liver in 1953...

Claims

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Application Information

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IPC IPC(8): G01N33/543
CPCG01N33/54326
Inventor 易龙糜漫天周曦刘蕾张玉郎和东
Owner ARMY MEDICAL UNIV
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