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A microsphere that utilizes buoyancy to separate and purify biological substances

A technology for separation and purification of biological substances, applied in the field of microspheres that use buoyancy to separate and purify biological substances, can solve the problems of cumbersome operation, complicated product equipment, high product price, etc., and achieve the effect of simple operation and rapid separation

Active Publication Date: 2021-03-19
厦门三一造血技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] There are the following problems in the separation of cells using magnetic bead separation: 1. The product device is complicated and requires a magnetic field generator; 2. The product is expensive, and the price of ordinary cell separation and separation device is over 10,000 RMB; Magnetic pole sorting, elution and other steps

Method used

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  • A microsphere that utilizes buoyancy to separate and purify biological substances
  • A microsphere that utilizes buoyancy to separate and purify biological substances
  • A microsphere that utilizes buoyancy to separate and purify biological substances

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Prepare antibody-linked silica microspheres by the following steps:

[0027] Step 1: Take a sample with a diameter of 50μm and a density of 0.8g / cm 3 The hollow silica microspheres were repeatedly washed 3 times with deionized water, and dried for later use;

[0028] Step 2: Take 5mg of the silica microspheres in step 1 and put them into 5ml SU-8 photoresist and mix them repeatedly, then take out the silica microspheres and place them in an oven at 95°C for 10 minutes to remove SU-8 Photoresist solvent;

[0029] Step 3: Expose the silica microbeads to 300W ultraviolet light for 450s. The distance between the microbeads and the ultraviolet light source is 15cm. Repeatedly wash 3 times with deionized water, blow dry with nitrogen, and store in a dark place for later use;

[0030] Step 4: Take 2ml of protein G solution with a solubility of 1mg / ml and 5mg of the silica microspheres prepared in step 2, react in a phosphate buffered saline solution with a pH of 10.5 at 8°C ...

Embodiment 2

[0036] Prepare antibody-linked silica microspheres by the following steps:

[0037] Step 1: Take a sample with a diameter of 30μm and a density of 0.6g / cm 3 The hollow silica microspheres were repeatedly washed 3 times with deionized water, and dried for later use;

[0038] Step 2: Take 5mg of the silica microspheres in step 1 and put them into 5ml SU-8 photoresist and mix them repeatedly, then take out the silica microspheres and place them in an oven at 95°C for 10 minutes to remove SU-8 Photoresist solvent;

[0039] Step 3: Expose the silica microbeads to 300W ultraviolet light for 450s. The distance between the microbeads and the ultraviolet light source is 10-15cm. Repeatedly wash 3 times with deionized water, blow dry with nitrogen, and store in a dark place for later use;

[0040] Step 4: Take 2ml of protein G solution with a solubility of 0.1mg / ml and 5mg of the silica microspheres prepared in step 2, react in a phosphate buffered saline solution with a pH of 9 at 0°...

Embodiment 3

[0046] Prepare antibody-linked silica microspheres by the following steps:

[0047] Step 1: Take a sample with a diameter of 40μm and a density of 0.7g / cm 3 The hollow silica microspheres were repeatedly washed 3 times with deionized water, and dried for later use;

[0048] Step 2: Take 5mg of the silica microspheres in step 1 and put them into 5ml SU-8 photoresist and mix them repeatedly, then take out the silica microspheres and place them in an oven at 95°C for 10 minutes to remove SU-8 Photoresist solvent;

[0049] Step 3: Expose the silica microbeads to 300W ultraviolet light for 450s. The distance between the microbeads and the ultraviolet light source is 10-15cm. Repeatedly wash with deionized water for 3 times, blow dry with nitrogen, and store in a dark place for later use;

[0050] Step 4: Take 2ml of protein G solution with a solubility of 0.5mg / ml and 5mg of the silica microspheres prepared in step 2, react in a phosphate buffered saline solution with a pH of 9.8...

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Abstract

The invention discloses a microsphere for separating and purifying biological substances by utilizing a buoyancy force, and belongs to the field of separation and purification of the biological substances. The microsphere comprises a silica microsphere of which the inner part is hollow, wherein the periphery of the silica microsphere is coated with a SU-8 photoresist; crosslinking of the SU-8 photoresist on the surface of the silica microsphere is realized under ultraviolet radiation; the silica microsphere modified by the SU-8 photoresist can be in ring-opening reaction under the situation that the PH (Potential of Hydrogen) is greater than 9.0 and can be linked with G protein, and the silica microsphere modified by the SU-8 photoresist can be further linked with an antibody under a condition that the PH is 7.45 so as to be used for separation and purification of target substances. By taking a separated cell as an example, the modified microsphere can be used for cell separation and has the following advantages that 1, rapid separation of target cells: the time is shortened to be 15 minutes and accounts for one fourth of the time of a traditional method; 2, simple operation: separation steps can be completed just by common instruments without a complicated device; 3, economy: the cost for obtaining high-purity cells in the same quantities accounts for one fourth of that of thetraditional method.

Description

technical field [0001] The invention belongs to the field of separation and purification of biological substances, in particular to a microsphere which utilizes buoyancy to separate and purify biological substances. Background technique [0002] Biological substances include cells, bacteria, nucleic acids, proteins, antibodies and other substances. Cells, as the basic constituent units of the human body and animals, are important basic raw materials for scientific research institutions and drug research institutions. Cells are used in almost all basic biological research and drug development, and the quality of cell products is directly related to the quality of the entire scientific research experiment and drug research. Cells often exist in tissues or blood. Accurate research on a specific cell requires the separation and purification of specific cells from different types of tissues and blood. Currently, the commonly used cell separation and purification method is the use...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J13/02C12N5/00
CPCB01J13/02C12N5/00
Inventor 叶永清黄能平
Owner 厦门三一造血技术有限公司