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
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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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