Purification method of plant exosome

A purification method and exosome technology, applied in biochemical equipment and methods, plant cells, cell dissociation methods, etc., can solve the problems of low separation and extraction, insufficient exosome activity, low purity, etc., and achieve simple operation. , Easy mass production, high purity effect

Pending Publication Date: 2022-05-27
广州远想医学生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a method for purifying plant exosomes, in order to overcome the technical problems of insufficient exosome activity, less separation and extraction, and low purity in the existing methods

Method used

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  • Purification method of plant exosome

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1: a kind of purification method of plant exosome

[0034] The specific steps of this method are as follows:

[0035] (1) Take Xiangyuan fruit and add crushing liquid (1×pbs) at 1:1000 (w / w);

[0036] (2) Collect the broken liquid, centrifuge at 3000rpm and 4°C for 30min, remove cell debris, and collect the supernatant;

[0037] (3) Add precipitation solution (precipitation solution: 10% PEG8000, 10% glycerol, balance water) to the supernatant, and place at 4°C for 10 hours;

[0038] (4) Centrifuge the mixed solution at 10000rpm and 4°C for 30min, discard the supernatant, collect the precipitate, add PBS (pH 7.0) to completely dissolve the precipitate, and obtain a lysate;

[0039] (5) separate the lysate by gel filtration chromatography to obtain exosomes; specifically:

[0040] The chromatographic column is Focurose 6FF, which is suitable for component separation and moderate purification of biological macromolecules. The particle size ranges from 45 to ...

Embodiment 2

[0044] Embodiment 2: a kind of purification method of plant exosome

[0045] The specific steps of this method are as follows:

[0046] (1) Take Xiangyuan fruit and add crushing liquid (1×pbs) at 1:1000 (w / w);

[0047] (2) Collect the broken liquid, centrifuge at 3000rpm and 4°C for 30min, remove cell debris, and collect the supernatant;

[0048] (3) Add precipitation solution (precipitation solution: 12% PEG8000, 15% glycerol, balance water) to the supernatant, and place at 4°C for 10 hours;

[0049] (4) Centrifuge the mixed solution at 10000rpm and 4°C for 30min, discard the supernatant, collect the precipitate, add PBS (pH 7.0) to completely dissolve the precipitate, and obtain a lysate;

[0050] (5) separate the lysate by gel filtration chromatography to obtain exosomes; specifically:

[0051] The chromatographic column is Focurose 6FF, which is suitable for component separation and moderate purification of biological macromolecules. The particle size ranges from 45 to ...

Embodiment 3

[0055] Embodiment 3: a kind of purification method of plant exosome

[0056] The specific steps of this method are as follows:

[0057] (1) Take Xiangyuan fruit and add crushing liquid (1×pbs) at 1:1000 (w / w);

[0058] (2) Collect the broken liquid, centrifuge at 3000rpm and 4°C for 30min, remove cell debris, and collect the supernatant;

[0059] (3) Add precipitation solution (precipitation solution: 10% PEG8000, 10% glycerol, balance water) to the supernatant, and place at 4°C for 10 hours;

[0060] (4) Centrifuge the mixed solution at 10000rpm and 4°C for 30min, discard the supernatant, collect the precipitate, add PBS (pH 7.0) to completely dissolve the precipitate, and obtain a lysate;

[0061] (5) separate the lysate by gel filtration chromatography to obtain exosomes; specifically:

[0062] The chromatographic column is Focurose 6FF, which is suitable for component separation and moderate purification of biological macromolecules. The particle size range is 45-165 μm...

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Abstract

The invention belongs to the technical field of exosome separation, discloses a purification method of plant exosomes, and aims to overcome the technical problems of insufficient exosome activity, small separation and extraction amount and low purity in the existing method. The purification method of the plant exosome comprises the following steps: collecting plant cell disruption liquid, and centrifugally collecting supernate; adding a precipitation solution into the supernate to obtain a mixed solution; centrifuging the mixed solution, discarding the supernatant, collecting the precipitate, and adding PBS to completely dissolve the precipitate so as to obtain a dissolved solution; and carrying out gel filtration chromatography separation on the dissolved solution to obtain the exosome. According to the method, separation and purification of large-scale and high-purity exosomes are achieved through combination of PEG8000 precipitation and gel filtration chromatography, the content, purity and biological activity of the exosomes can be guaranteed at the same time, and the separated exosomes are high in yield and purity; the method is high in efficiency, simple to operate and easy for large-scale production.

Description

technical field [0001] The invention relates to the technical field of exosome separation, in particular to a method for purifying plant exosomes. Background technique [0002] Exosomes are tiny vesicles (30-150 nanometers) secreted by stem cells. They resemble intercellular "signals" that can transfer DNA, RNA or proteins between cells, thereby affecting the function of recipient cells. [0003] At present, exosomes are widely used. In stem cell and regenerative medicine, exosomes are used to treat diseases ranging from heart disease to respiratory disease. In the field of skin care, when exosomes enter the skin, they can automatically release nutrients, continuously improve the cell environment, and repair skin problems; stimulate cell migration, proliferation and collagen synthesis, increase skin thickness, and repair the barrier; it can enhance the expression of antioxidant enzymes, thereby Enhances skin's antioxidant capacity; repairs cell aging caused by UV rays and...

Claims

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Application Information

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IPC IPC(8): C12N5/04
CPCC12N5/04C12N2509/10
Inventor 邹衡芳周晗陈宁赖云来昌冲
Owner 广州远想医学生物技术有限公司
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