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Pilot-scale extraction method of plant-derived exosome and application of plant-derived exosome

A plant source and extraction method technology, applied in the field of plant source exosome extraction and enrichment purification, can solve the problems of limiting the application of plant exosomes, lack of efficient extraction methods of plant exosomes, etc., and achieve good application prospects and research Value, good cosmetic or therapeutic effect, effect of efficient extraction

Pending Publication Date: 2022-02-08
上海卡替睿舒医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still a lack of efficient extraction methods for plant exosomes, which limits the further application of plant exosomes.

Method used

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  • Pilot-scale extraction method of plant-derived exosome and application of plant-derived exosome
  • Pilot-scale extraction method of plant-derived exosome and application of plant-derived exosome
  • Pilot-scale extraction method of plant-derived exosome and application of plant-derived exosome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0127] Lily exosome preparations: Example 1

[0128] Weigh fresh lily (including stem, leaves, flowers) 1 kg, washed three times with tap water, then rinsed three times with purified water, dry. Chopped, added juicer, was added 2000mL PBS buffer, homogenized 2 times. The residue was filtered with gauze large. Placed in shake flasks, shaken at a shaker for 24 hours, the temperature is 22 ℃, rotation speed of 100rpm. The mixture was centrifuged using a centrifuge 5000rpm 30min, the supernatant was collected. The supernatant was 0.45um hollow fiber ultrafiltration column, and then the filtrate was again 500kDa hollow fiber ultrafiltration column was concentrated to 300mL. Was added 75mL of PEG8000 / NaCl, mixed and left for 24 hours within a refrigerator at 4 ℃. 4000rpm, 4 ℃, centrifuged 10min, the precipitate was collected, resuspended with 30mL saline, filter sterilized 0.22um filter. Placed in -80 ℃ refrigerator storage. Protein concentrations were determined by Bradford method, t...

Embodiment 2

[0135] Seabuckthorn exosome preparations: Example 2

[0136] Weigh 2 kg of fresh sea buckthorn, washed 3 times with tap water, then rinsed three times with purified water, dry. Added juicer, was added 4000 mL PBS buffer, homogenized 3 times. The residue was filtered with gauze large. Placed in shake flasks, shaken at a shaker for 24 hours, the temperature is 25 ℃, speed of 120rpm. The mixture was centrifuged using a centrifuge 5000rpm 30min, the supernatant was collected. The supernatant was 0.45um hollow fiber ultrafiltration column, and then the filtrate was again 300kDa hollow fiber ultrafiltration column was concentrated to 400mL. Was added 100mL of PEG8000 / NaCl, mixed and left for 24 hours within a refrigerator at 4 ℃. 4000rpm, 4 ℃, centrifuged 15min, the precipitate was collected, resuspended with 40mL saline, filter sterilized 0.22um filter. Placed in -80 ℃ refrigerator storage. Protein concentrations were determined by Bradford method, to calculate the total amount of ex...

Embodiment 3

[0143] Example 3: Cistanche exosome preparations

[0144] Cistanche weighed 1 kg of dried, washed 3 times with tap water, then rinsed three times with purified water, dry. Added juicer, was added 5000mL PBS buffer, homogenized 3 times. The residue was filtered with gauze large. Placed in shake flasks, shaken at a shaker for 48 hours to a temperature of 24 ℃, rotation speed of 100rpm. The mixture was centrifuged using a centrifuge 5000rpm 30min, the supernatant was collected. The supernatant was 0.45um hollow fiber ultrafiltration column, and then the filtrate was again 300kDa hollow fiber ultrafiltration column was concentrated to 500mL. Was added 125mL of PEG8000 / NaCl, mixed and left for 24 hours within a refrigerator at 4 ℃. 4000rpm, 4 ℃, centrifuged 20min, the precipitate was collected, resuspended with 50mL saline, filter sterilized 0.22um filter. Placed in -80 ℃ refrigerator storage. Protein concentrations were determined by Bradford method, to calculate the total amount of...

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Abstract

The invention belongs to the field of biotechnology and bioengineering, and discloses a method for extracting exosomes from plants, wherein the method comprises the steps: taking any part of a plant, crushing, then adding a PBS buffer solution for soaking and permeating treatment to release the exosomes in the plants into a buffer solution, centrifuging and collecting the supernatant; and then respectively carrying out microfiltration and ultrafiltration concentration through a hollow fiber column to obtain required exosomes, wherein the soaking and permeating treatment is carried out on a shaking table in a shaking manner, the temperature is 22-25 DEG C, and the rotating speed is 90-120 rpm. According to the method for extracting the exosomes, the exosomes in the plants can be efficiently extracted, the extraction steps are few, the extraction time is short, the content of the obtained exosomes is high, and the method has the advantages of being simple in post-treatment, capable of achieving large-scale production and the like.

Description

Technical field [0001] The present invention belongs to the field of biotechnology and bioengineering, relates to a general method of extracting a plant-derived exosomes purification and enrichment, the production method can be used to amplify. Background technique [0002] Recent studies have found that plants, secreted vesicles bodies, 40-100 nm in diameter, named exosome, comprising a protein, mRNA, miRNA and other substances, has an important function information flow and material transport between cells. Exosomes has been widespread concern for the natural therapy vectors, also with a clear reduce the extent of tissue damage and promoting damaged tissue morphology and functional repair of school. E.g. exosomes Arabidopsis leaves contain a variety of antioxidants, and the like may be used in cosmetic products. [0003] Conventional extraction plant exosomes from primary method with sucrose density gradient ultra-centrifugation of the juice, the method comprising preparing ult...

Claims

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Application Information

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IPC IPC(8): C12N5/04A61K8/99A61K8/9794A61K8/9789A61Q19/08A61K36/8967A61K36/185A61K36/64A61P39/06
CPCC12N5/04A61K8/99A61K8/9794A61K8/9789A61Q19/08A61K36/8967A61K36/185A61K36/64A61P39/06
Inventor 李军
Owner 上海卡替睿舒医学检验实验室有限公司
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