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Method for extracting extracellular vesicles from milk

A technology of cells and vesicles, applied in the field of extracting extracellular vesicles, can solve the problems of high cost, low yield and low efficiency of isolated EVs, and achieve the effects of low cost, great clinical transformation prospects, and increased production

Active Publication Date: 2021-08-06
PEKING UNIV FIRST HOSPITAL
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Therefore, the technical problem to be solved by the present invention is to overcome the defects of low yield, low efficiency and high cost of isolated EVs in the prior art, thereby providing a method for extracting extracellular vesicles from milk

Method used

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  • Method for extracting extracellular vesicles from milk
  • Method for extracting extracellular vesicles from milk
  • Method for extracting extracellular vesicles from milk

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] 1. Milk Collection and Whey Preparation

[0044] 1.1 Pasteurized raw milk produced in the market for refrigerated storage and transportation on the day of purchase—skimmed milk (Beijing Sanyuan Food Co., Ltd.);

[0045] 1.2 Take 50mL skimmed milk in a test tube and preheat at 37°C for 10 minutes;

[0046]1.3 Acidified milk: add a little 6mol / L hydrochloric acid, the volume ratio of milk / hydrochloric acid is about 100:1, adjust the pH to 4.6, and let stand for 5 minutes;

[0047] 1.4 Centrifuge at 5000xg for 1 hour at 22°C to remove casein and obtain a relatively transparent supernatant—whey;

[0048] 1.5 Add powdered ammonium sulfate solids to the supernatant obtained in the previous step and mix thoroughly, and prepare solutions with different saturations of ammonium sulfate. This experiment adopts the following table (the solution of the remaining saturation of ammonium sulfate is calculated using the following website: https: / / www.encorbio.com / protocols / AM-SO4....

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Abstract

The invention relates to the technical field of biology, in particular to a method for extracting extracellular vesicles from milk. The invention provides a method for extracting extracellular vesicles from milk. The method comprises the step of adding ammonium sulfate into whey. The method for extracting the extracellular vesicles from the milk has the following advantages that it is verified for the first time that the ammonium sulfate can be used for precipitation of mEVs, and the mEVs still have activity after redissolution; the method can increase the yield of the mEVs, can achieve the equivalent purity, is lower in cost, and can be used for large-scale production of the mEVs; and in the future, the mEVs can be used as natural nano-carriers to deliver bioactive molecules such as proteins, medicines and functional RNA, can regulate the functions of cells and treat diseases, and has a great clinical transformation prospect.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for extracting extracellular vesicles from milk. Background technique [0002] Extracellular vesicles (EVs) are nano-spherical substances with a bilayer membrane structure and biological activity actively secreted by cells, mainly composed of lipids, proteins and genetic material. Its main types include: exosomes (particle size 30-150nm), microvesicles (microvesicles, particle size 100-1000nm), apoptotic bodies (apoptotic bodies, particle size > 1000nm). EVs are characterized by small molecular structure and high biocompatibility, and can be used to transport substances such as lipids, proteins, DNA, and RNA, and are natural endogenous nanocarriers. EVs widely exist in organisms, including plants, mammals and humans. In principle, all living cells secrete EVs. Currently, EVs are mainly extracted from cell culture medium and body fluids such as blood, urine, cerebrospin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N15/87A61K47/46
CPCC12N5/0631C12N15/87A61K47/46C12N2509/00
Inventor 关瑞礼谭晓辉方冬李学松周利群
Owner PEKING UNIV FIRST HOSPITAL
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