A method of extracting milk exosomes

Abstract

The present invention provides a method for extracting milk exosomes, the method comprising the following steps: (1) taking a milk sample, centrifuging at 2000g, 4°C, and 30min, and collecting the supernatant A; (2) collecting the collected Supernatant A was centrifuged at 12000g, 4°C, 30min, and supernatant B was collected; (3) The collected supernatant B was further adjusted to pH 5.5-6.0; (4) Under the condition of 36-37°C water bath, Adding chymosin to the supernatant B after adjusting the pH to precipitate milk protein to obtain supernatant C; (5) freeze-drying the subcomponents of supernatant C that can pass through a 0.22 μM membrane into powder, namely for milk exosomes. Compared with the traditional centrifugal-ultrahigh-speed centrifugation method, the method for extracting milk exosomes of the present invention reduces the steps of the ultrahigh-speed centrifugation method, saves the cost of consumables used in extraction, and greatly increases the sample extraction volume.

Description

technical field [0001] The invention belongs to the field of biotechnology methods, and in particular relates to a method for extracting milk exosomes. Background technique [0002] Exosomes are a type of membrane-bound vesicles formed by endocytosis, which are released into the extracellular environment by fusion of multivesicular bodies (MVBs) and appear round or cup-shaped under a transmission electron microscope. Phospholipid bilayer structure with a diameter of 40-100nm. Studies have shown that different cells in vivo and in vitro can secrete exosomes under physiological or pathological conditions. In addition, exosomes are also found in various body fluids and breast milk of animals or in vivo. And it has also been proved that the materials in the exosome are closely related to the origin and production conditions of cells. Studies have shown that mRNA and miRNA are found in milk exosomes of humans, cows and pigs and can be transported into immune cells, potentially...

AI Technical Summary

Problems solved by technology

Known information shows: 1) The kit method, the extraction sample volume is small, the price is expensive, and the extraction cost is high; 2) The density gradient centrifugation method is cumbersome, takes a long time, and has a large loss; 3) The ultra-speed exosome method obtains The rate is higher, and the steps are simpler than the density gradient, but its purity is very controversial

Except for the kit method that does not require an ultra-high-speed centrifuge and can complete the extraction steps in an ordinary molecular laboratory, the other two methods need to rely on this relatively specific instrument, and the amount of single sample extraction is too small ; It is difficult to achieve mass production or large-volume trials

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