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Supercritical filtration affinity adsorption system, construction method thereof, ultramicro factor preparation method and application

An adsorption system and supercritical technology, applied in the field of biomedicine, can solve the problems of small sample size, poor repeatability, and inability to prepare ultrafine factors with large-scale GMP grade.

Pending Publication Date: 2021-07-09
京美瑞禾健康科技(辽宁)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] For this reason, the present invention provides a supercritical filtration affinity adsorption system and its construction method and ultramicro factor preparation method and applicat

Method used

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  • Supercritical filtration affinity adsorption system, construction method thereof, ultramicro factor preparation method and application
  • Supercritical filtration affinity adsorption system, construction method thereof, ultramicro factor preparation method and application
  • Supercritical filtration affinity adsorption system, construction method thereof, ultramicro factor preparation method and application

Examples

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Embodiment 1

[0044] Example 1 The present invention provides a method for extracting and purifying exosomes using a double tangential flow filtration system

[0045] A structural diagram of a double tangential flow filtration system of the present embodiment, see Figure 4 .

[0046] 1. Umbilical cord mesenchymal stem cell culture

[0047] 1. Extraction and preparation of umbilical cord mesenchymal stem cells

[0048] Collect the newborn umbilical cord from the operating room, rinse the umbilical cord with sterile PBS buffer, remove the blood vessels with scissors and tweezers, peel off the Wahrenheit tissue inside, and cut the tissue to 1mm 3 The size of the organization block. Spread the tissue pieces evenly in the culture dish, and cultivate them in the incubator for 1 h (the culture condition is 5% CO 2 , 37°C). When the tissue block is firmly attached to the wall, add an appropriate amount of stem cell culture medium and place it in an incubator for culture (the culture condition i...

Embodiment 2

[0062] The extraction and preparation of embodiment 2 ultrafine factors

[0063] This example figure 1 Flowchart for preparation of ultrafine factor solution and figure 2 The structure diagram of the nanofactor solution preparation shown

[0064] (1) Add sterile water to the first storage tank 1 and the second storage tank 2, open the peristaltic pumps (3 and 4) respectively, and flush the system;

[0065] (2) Add the mixed solution in the first storage tank 1, open the first peristaltic pump 4 to circulate the system, and the mixed sample solution flows through the supercritical filter membrane device provided with the supercritical filter membrane, so that the solution containing ultrafine factors Enter in the second storage tank 2; when the liquid volume in the first storage tank 1 is the minimum operating volume, add 500ml sterile PBS buffer solution and continue to concentrate and filter to the minimum operating volume;

[0066] (3) When the liquid volume in the secon...

Embodiment 3

[0068] Example 3 A system for extracting and purifying ultramicrofactors

[0069] Described a kind of supercritical filtration affinity adsorption system that is used for extraction of microfactor includes first storage tank 1, second storage tank 2, supercritical filtration membrane device 5, affinity adsorption device 6, first peristaltic pump 3, The second peristaltic pump 4, switch K1 and switch K2;

[0070] Wherein, the outlet of the first storage tank is connected to the inlet of the supercritical filter device through the first peristaltic pump; the outlet of the supercritical filter device is connected to the inlet of the storage tank; the filtered liquid outlet is connected to the first inlet of the second storage tank; The outlet of the second storage tank is connected to the affinity adsorption system through the second peristaltic pump; the affinity adsorption device is respectively connected to the second inlet of the second storage tank through the switch K2, and...

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Abstract

The invention discloses a supercritical filtration affinity adsorption system and a construction method thereof as well as an ultramicro factor preparation method and application. The system comprises a first storage tank, a first peristaltic pump, a supercritical filtering device, a second storage tank, a second peristaltic pump and an affinity adsorption device. The method comprises the following steps: pretreating a sample solution; extracting and purifying exosomes, crushing the exosomes, mixing a crushed exosome solution with a non-exosome solution, and putting the mixture into a first storage tank; subjecting a solution filtered by a supercritical filtering device to adsorption and elution by an affinity adsorption device to obtain an ultramicro factor solution. The prepared ultramicro factor is applied to preparation of biological products related to the ultramicro factor. The method can be used for preparing large-scale ultramicro factors, can be used for extracting ultramicro factors from various sample sources, not only can remove non-ultramicro factor substances with large particles in samples, but also can extract and concentrate the non-ultramicro factor substances to the greatest extent, and is suitable for producing and preparing the large-scale ultramicro factors.

Description

[0001] The invention relates to the technical field of biomedicine, and relates to a supercritical filtration affinity adsorption system, a construction method thereof, and a preparation method and application of ultramicrofactors. Background technique [0002] Ultramicrofactors are active ingredients secreted by living cells such as plants, animals, and microorganisms, with a molecular weight of less than 5KD. It comes from a wide range of sources, including tissue fluid, culture fluid, somatic cells, stem cells, and cells that have been induced to differentiate. According to the source, it can be divided into plant ultrafine factors, microbial ultrafine factors, stem cell ultrafine factors, body fluid ultrafine factors, tissue fluid ultrafine factors, somatic cell ultrafine factors, etc. [0003] Ultramicrofactors contain more than 1,000 active ingredients such as small molecule polypeptides, lipopeptides, lipids, phospholipids secreted by cells, microRNA and various factors...

Claims

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Application Information

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IPC IPC(8): B01D61/14B01D15/10B01L3/00B01D36/04C07K1/22
CPCB01D61/147B01D15/10B01L3/502753B01D36/045C07K1/22Y02P20/54
Inventor 张海心
Owner 京美瑞禾健康科技(辽宁)有限公司
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