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KR-102962026-B1 - METHOD FOR PRODUCING EXOSOMES DERIVED FROM NATURAL PRODUCTS AND ANIMALS

KR102962026B1KR 102962026 B1KR102962026 B1KR 102962026B1KR-102962026-B1

Abstract

The present disclosure relates to a method for producing exosomes derived from natural products and animals, and more specifically, to a method for isolating and purifying exosomes from raw materials such as medicinal plants, stem cells, and milk.

Inventors

  • 문웅식
  • 김미경
  • 문찬영
  • 문요성

Assignees

  • (주)마이크로 사이언스 테크

Dates

Publication Date
20260507
Application Date
20241226

Claims (16)

  1. A method for producing exosomes from natural products and animal-derived substances, A cleaning process for cleaning and softening raw materials using an ultrasonic cleaner equipped with an electronic vibrator; A drying process for freeze-drying washed raw materials using a cryogenic freeze dryer utilizing liquid nitrogen, carbon dioxide dry ice, or a refrigerant; A grinding process for grinding dried raw materials using a super-particle grinding system; A separation and purification process comprising mixing one or more solvents selected from the group consisting of ethyl alcohol, purified water, isopropyl alcohol, ethylene glycol, butyl alcohol, propylene glycol, and butylene glycol in a weight ratio of 10 to 100 times relative to the raw material, and separating and purifying exosomes from the ground raw material using a roll-to-roll cast cellulose acetate membrane filter; and A method for manufacturing exosomes comprising a packaging process for packaging in a sterile state by passing the above-described separated and purified exosome solution through a multi-stage membrane filter system, wherein the first membrane filter has pores of 50 to 150 μm and the second membrane filter has pores of 0.1 to 1.0 μm. The above raw material is selected from peony leaves, peony roots, lotus flowers, lotus leaves, and lotus stems and their calluses, and A method for producing exosomes, characterized in that the above separation and purification process is performed at a temperature of 10℃ to 150℃.
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  16. A method for extracting, isolating, and preparing mucin and choline from exosomes prepared by the method of claim 1.

Description

Method for producing exosomes derived from natural products and animals The present disclosure relates to a method for producing exosomes derived from natural products and animals, and more specifically, to a method for isolating and purifying exosomes from raw materials such as medicinal plants, stem cells, and milk. Exosomes are nano-sized extracellular vesicles secreted by cells that contain various biomaterials such as proteins, lipids, and nucleic acids, and have recently been attracting attention in various fields including cosmetics, pharmaceuticals, and health functional foods. In particular, natural and animal-derived exosomes are evaluated as having very high industrial value due to their excellent biocompatibility and low toxicity. Currently, manufacturing methods for the isolation and purification of exosomes, such as ultracentrifugation, exosome size-based separation, and exosome precipitation, are known. Ultracentrifugation is a method of isolating exosomes through centrifugation, and while it is widely used on a laboratory scale, it has several limitations for industrial production. Not only are ultracentrifuges, which are equipment for mass production, expensive, but there are also disadvantages such as a lower manufacturing yield compared to continuous manufacturing processes due to the single-stage process, and increased construction and facility costs when building mass production facilities. Exosome size-based separation is a method of separating exosomes using size exclusion chromatography or filters. While it has the advantage of being able to selectively separate exosomes of a specific size, it has limited processing capacity and suffers from reduced efficiency due to clogging. Additionally, exosome precipitation is a method of precipitating exosomes using polymers or salts. Although it allows for simple separation of exosomes, it has the problem of low purity and the potential for a large amount of impurities to be included. These existing manufacturing methods commonly have the disadvantage of low production yield relative to cost, as the separation and manufacturing processes for producing large quantities of exosomes require long working hours. Additionally, there are issues with difficult quality control of exosomes and difficulty in maintaining quality consistency during mass production. In particular, for exosomes derived from natural products, it is necessary to establish optimized conditions for each process step, from the pretreatment of raw materials to the final purification stage, but existing manufacturing methods fail to meet these requirements. In the case of animal-derived exosomes, ensuring sterility and maintaining stability are emerging as important challenges, but current manufacturing methods make it difficult to effectively solve these problems. Therefore, there is an urgent need to develop new manufacturing methods that can efficiently produce natural and animal-derived exosomes on an industrial scale. The following description of the present disclosure is not limited to specific embodiments and may be subject to various modifications and have various embodiments. Furthermore, it should be understood that the content described below includes all modifications, equivalents, and substitutions that fall within the spirit and scope of the present disclosure. In the following description, terms such as "first," "second," etc., are used to describe various components and are not limited in their meaning; they are used solely for the purpose of distinguishing one component from another. Additionally, the same reference numerals used throughout this specification indicate the same component. Unless otherwise stated in this specification, a specific step or process may be carried out at room temperature. Room temperature may be in the range of 15 to 30 degrees Celsius, and preferably in the range of 20 to 25 degrees Celsius. Singular expressions used in this disclosure include plural expressions unless the context clearly indicates otherwise. Furthermore, terms such as “comprising,” “having,” or “having,” as used below, should be interpreted as indicating the existence of the features, numbers, steps, actions, components, parts, or combinations thereof described in the specification, and should be understood as not precluding the existence or addition of one or more other features, numbers, steps, actions, components, parts, or combinations thereof. The terms or words used in this specification and claims are not intended to be interpreted as being limited to their commonly accepted dictionary meanings, and a person skilled in the art will clearly understand that said terms or words are used within the scope of conveying what this disclosure means, within the scope of expressing the concept that this specification and claims obviously intend to convey. Furthermore, a person skilled in the art will clearly understand that the configurations described in the aspects, mode