Method for purifying and concentrating composite biological active factors and concentrated liquid thereof

A technology of bioactive factors and growth factors, which is applied in the field of purification and concentration of compound bioactive factors and its concentrate, can solve the problems of inability to guarantee the effective activity of compound active factors, high cost of concentration and extraction, poor utilization rate, etc., and achieve delay Effect of skin aging, guaranteed nutrition and functional ingredients, low cost

Inactive Publication Date: 2019-07-19

雷艳

0 Cites 0 Cited by

AI-Extracted Technical Summary

Problems solved by technology

[0003] With the development of society, the use of compound active factors in medical treatment and skin care is getting higher and higher, and the extraction, concentration and preparation of concentrates containing compound active factors generally require the concentration and extraction ...

Abstract

The present invention discloses a method for purifying and concentrating composite biological active factors and concentrated liquid thereof. An organic solvent is added into biological tissue cultureliquid containing the composite biological active factors, the biological tissue culture liquid is continuously stirred to be uniform to obtain biological tissue mixed liquid, the obtained biologicaltissue mixed liquid is put into a pressure processor, a temperature-rising treatment is conducted, rapid depressurizing is conducted to normal pressure, besides, the biological tissue mixed liquid istaken out, centrifuging is conducted, after the centrifuging, filtration is conducted to remove residual biological tissue impurities in the biological tissue mixed liquid to obtain clarified liquidof the biological tissue mixed liquid, besides, an ultrafiltration membrane is used to filter out the clarified liquid to obtain a biological tissue ultrafiltrate containing the active factors, and besides, the ultrafiltrate is further purified and concentrated to obtain the concentrated liquid containing the composite biological active factors. The concentrated liquid containing the composite biological active factors can accelerate repair after a skin surgery and also has functions of smoothing fine lines, delaying skin aging, etc.

Application Domain

Colony-stimulating factorPeptide preparation methods +9

Technology Topic

UltrafiltrationChemistry +11

Examples

Experimental program(3)

Example Embodiment

[0019] Example 1

[0020] The present invention provides a method for purifying and concentrating composite biologically active factors and a concentrated solution thereof, including the following steps:

[0021] S1, adding the biological tissue culture medium containing the compound biologically active factors to the organic solvent, and continuously stirring until uniform, to obtain the biological tissue mixture;

[0022] S2: Put the biological tissue mixture obtained in S1 into a pressure processor, perform a pressure increase treatment, pressurize to 120 MPa, and hold for 5 minutes, then quickly release the pressure to normal pressure, and take out the biological tissue mixture;

[0023] S3: Add the biological tissue mixture solution taken out in S2 to the centrifuge and perform centrifugation. After the centrifugation is completed, filter through a 100nm filter medium to remove residual biological tissue impurities in the biological tissue mixture solution to obtain a biological tissue mixture The clear liquid of the solution;

[0024] S4: Take out the clear liquid of the biological tissue mixed solution obtained in S3, and use an ultrafiltration membrane to filter out organic macromolecular substances with a molecular weight greater than 3000 Daltons in the clear liquid to obtain an ultrafiltrate of biological tissues containing active factors;

[0025] S5: The ultrafiltrate obtained in S4 is subjected to nanofiltration technology for sub-concentration treatment to remove the organic solvent contained in the ultrafiltrate, and the ultrafiltrate is purified and concentrated again to obtain a concentrated solution containing complex biologically active factors .

[0026] Specifically, the compound biologically active factors include: human epidermal growth factor HEGF, vascular endothelial growth factor VEGF, fibroblast growth factor FGF, transforming growth factor ~ β1 TGF ~ β1, transforming growth factor ~ β2 TGF ~ β2, insulin-like No. 1 Growth factor IGF~1, insulin-like growth factor II, IGF~2, keratinocyte growth factor KGF, platelet-derived growth factor PDGF, erythrocyte growth factor EPO, colony stimulating factor CS and interleukin growth factors.

[0027] Specifically, in S1, the biological tissue culture medium is a frozen tissue-conditioned medium containing composite biologically active factors, which is used after being thawed.

[0028] Specifically, in S1, the organic solvent is any one or a combination of methanol, ethanol, isopropanol, n-butanol, acetone, methyl acetate, and ethyl acetate.

[0029] Specifically, in S2, the time for pressure boosting is controlled at 2 minutes, and the time for pressure relief is controlled within 5 seconds.

[0030] Specifically, in S3, the centrifugation speed of the biological tissue mixed solution in the centrifuge is 5000 rpm, and the centrifugation time is 10.

[0031] Specifically, in S5, the nanofiltration membrane of 300 Dalton is selected according to the molecular weight of the organic solvent in the ultrafiltrate in S4.

Example Embodiment

[0032] Example 2

[0033] A method for purification and concentration of compound biologically active factors and a concentrated solution thereof include the following steps:

[0034] S1, adding the biological tissue culture medium containing the compound biologically active factors to the organic solvent, and continuously stirring until uniform, to obtain the biological tissue mixture;

[0035] S2: Put the biological tissue mixture obtained in S1 into a pressure processor, perform pressure-increasing treatment, pressurize to 400MPa, and hold for 8 minutes, then quickly release the pressure to normal pressure, and take out the biological tissue mixture;

[0036] S3: Add the biological tissue mixture solution taken out in S2 to the centrifuge and perform centrifugation. After the centrifugation is completed, filter through a 100nm filter medium to remove residual biological tissue impurities in the biological tissue mixture solution to obtain a biological tissue mixture The clear liquid of the solution;

[0037] S4: Take out the clear liquid of the biological tissue mixed solution obtained in S3, and use an ultrafiltration membrane to filter out organic macromolecular substances with a molecular weight greater than 4000 Daltons in the clear liquid to obtain an ultrafiltrate of biological tissue containing active factors;

[0038] S5: The ultrafiltrate obtained in S4 is subjected to nanofiltration technology for sub-concentration treatment to remove the organic solvent contained in the ultrafiltrate, and the ultrafiltrate is purified and concentrated again to obtain a concentrated solution containing complex biologically active factors .

[0039] Specifically, the compound biologically active factors include: human epidermal growth factor HEGF, vascular endothelial growth factor VEGF, fibroblast growth factor FGF, transforming growth factor ~ β1 TGF ~ β1, transforming growth factor ~ β2 TGF ~ β2, insulin-like No. 1 Growth factor IGF~1, insulin-like growth factor II, IGF~2, keratinocyte growth factor KGF, platelet-derived growth factor PDGF, erythrocyte growth factor EPO, colony stimulating factor CS and interleukin growth factors.

[0040] Specifically, in S1, the biological tissue culture medium is a frozen tissue-conditioned medium containing composite biologically active factors, which is used after being thawed.

[0041] Specifically, in S1, the organic solvent is any one or a combination of methanol, ethanol, isopropanol, n-butanol, acetone, methyl acetate, and ethyl acetate.

[0042] Specifically, in S2, the time for pressure boosting is controlled at 3 minutes, and the time for pressure relief is controlled within 7 seconds.

[0043] Specifically, in S3, the centrifugal speed of the biological tissue mixed solution in the centrifuge is 6000 rpm, and the centrifugal time is 15 minutes.

[0044] Specifically, in S5, the nanofiltration membrane of 450 Dalton is selected according to the molecular weight of the organic solvent in the ultrafiltrate in S4.

Example Embodiment

[0045] Example 3

[0046] A method for purification and concentration of compound biologically active factors and a concentrated solution thereof include the following steps:

[0047] S1, adding the biological tissue culture medium containing the compound biologically active factors to the organic solvent, and continuously stirring until uniform, to obtain the biological tissue mixture;

[0048] S2: Put the biological tissue mixture obtained in S1 into a pressure processor, perform pressure-increasing treatment, pressurize to 700 MPa, and hold for 10 minutes, quickly release the pressure to normal pressure, and take out the biological tissue mixture;

[0049] S3: Add the biological tissue mixture solution taken out in S2 to the centrifuge and perform centrifugation. After the centrifugation is completed, filter through a 100nm filter medium to remove residual biological tissue impurities in the biological tissue mixture solution to obtain a biological tissue mixture The clear liquid of the solution;

[0050] S4: Take out the clear liquid of the biological tissue mixed solution obtained in S3, and use an ultrafiltration membrane to filter out the organic macromolecular substances with a molecular weight greater than 5000 Daltons in the clear liquid to obtain an ultrafiltrate of biological tissues containing active factors;

[0051] S5: The ultrafiltrate obtained in S4 is subjected to nanofiltration technology for sub-concentration treatment to remove the organic solvent contained in the ultrafiltrate, and the ultrafiltrate is purified and concentrated again to obtain a concentrated solution containing complex biologically active factors .

[0052] Specifically, the compound biologically active factors include: human epidermal growth factor HEGF, vascular endothelial growth factor VEGF, fibroblast growth factor FGF, transforming growth factor ~ β1 TGF ~ β1, transforming growth factor ~ β2 TGF ~ β2, insulin-like No. 1 Growth factor IGF~1, insulin-like growth factor II, IGF~2, keratinocyte growth factor KGF, platelet-derived growth factor PDGF, erythrocyte growth factor EPO, colony stimulating factor CS and interleukin growth factors.

[0053] Specifically, in S1, the biological tissue culture medium is a frozen tissue-conditioned medium containing composite biologically active factors, which is used after being thawed.

[0054] Specifically, in S1, the organic solvent is any one or a combination of methanol, ethanol, isopropanol, n-butanol, acetone, methyl acetate, and ethyl acetate.

[0055] Specifically, in S2, the time for boosting processing is controlled at 5 minutes, and the time for pressure relief is controlled within 10s.

[0056] Specifically, in S3, the centrifugation speed of the biological tissue mixed solution in the centrifuge is 7000 rpm, and the centrifugation time is 20 min.

[0057] Specifically, in S5, the nanofiltration membrane is 600 Daltons according to the molecular weight of the organic solvent in the ultrafiltrate in S4.

PUM

Description & Claims & Application Information

We can also present the details of the Description, Claims and Application information to help users get a comprehensive understanding of the technical details of the patent, such as background art, summary of invention, brief description of drawings, description of embodiments, and other original content. On the other hand, users can also determine the specific scope of protection of the technology through the list of claims; as well as understand the changes in the life cycle of the technology with the presentation of the patent timeline. Login to view more.

Composition of phellinus igniarius submerged fermentation concentrated solution and thallus extract as well as preparation method and application of composition

PendingCN114010548AStimulates collagen productiondelay skin aging

Owner:TIANJIN KANGTING BIOLOGICAL ENG GRP CO LTD

Silk mask with acne removal and skin care efficacy and preparation method of silk mask

InactiveCN106511164Amild performancedelay skin aging

Owner:JIANGSU AIXISHI TECH SERVICE CONSULTATION CO LTD

Resveratrol component contained cosmetic

PendingCN110538081Adelay skin agingPromote melanin metabolism

Owner:安婕妤化妆品科技股份有限公司

Classification and recommendation of technical efficacy words

Promote nutrient metabolism

Orthopedic nursing device with infrared function

PendingCN108478390APromote nutrient metabolismAccelerated blood lymphatic circulation

Owner:THE SECOND PEOPLES HOSPITAL OF NANTONG

Method for purifying and concentrating composite biological active factors and concentrated liquid thereof

AI-Extracted Technical Summary

Problems solved by technology

Abstract

Examples

Example Embodiment

Example Embodiment

Example Embodiment

PUM

Description & Claims & Application Information

Similar technology patents

Anti-aging skin care product composition and preparation method thereof

Orthopedic nursing device with infrared function

Moringa oil facial mask and making method thereof

Composition of phellinus igniarius submerged fermentation concentrated solution and thallus extract as well as preparation method and application of composition

Silk mask with acne removal and skin care efficacy and preparation method of silk mask

Resveratrol component contained cosmetic

Classification and recommendation of technical efficacy words

Orthopedic nursing device with infrared function