Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Medicine or cosmetic prepared from fibroblast extracellular vesicles

A technology of vesicles and monoclonal antibodies, applied in cosmetic preparations, animal cells, vertebrate cells, etc., can solve the problems of easy aging of fibroblasts, low activity of exosomes, low yield, etc., and achieve good market application value , good effect of anti-aging activity

Active Publication Date: 2022-07-12
诺赛联合(北京)生物医学科技有限公司
View PDF9 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current problem in the existing technology is that fibroblasts are prone to aging during the culture process, resulting in low activity and low yield of the prepared exosomes. Therefore, improving the activity of fibroblasts is the key direction of research

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Medicine or cosmetic prepared from fibroblast extracellular vesicles
  • Medicine or cosmetic prepared from fibroblast extracellular vesicles
  • Medicine or cosmetic prepared from fibroblast extracellular vesicles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1P53

[0037] Example 1 Preparation of P53 monoclonal antibody

[0038] Recombinant human p53 protein (abcam, Cat. No. ab84768, sequence shown in SEQ ID NO: 1) was used as the immunogen. The recombinant protein was thoroughly mixed with an equal volume of Freund's complete adjuvant, and BALB / c mice were subcutaneously injected with 50 μg / mice of the recombinant protein. Four weeks after the initial immunization, multiple subcutaneous injections on the back were given to boost the immunization, and 4 weeks later, the immunization was boosted again, with 50 μg of the immunogen per animal each time. Seven to ten days after the third immunization, blood was collected from the tail vein to measure the serum antibody titer by ELISA. Four days before fusion, the mouse with the highest antibody titer was selected and boosted by intraperitoneal injection of 150 μg of unadjuvanted antigen.

[0039] Aseptically take spleen cells of booster immunized BALB / c mice, use 50% polyethylene glycol as...

Embodiment 22

[0041] Example 2 Affinity, isotype identification, sequence analysis and specificity identification of 2F6 monoclonal antibody

[0042] Using the AMC sensor, the purified 2F6 antibody was diluted with PBST to 10ug / ml, and the recombinant protein was diluted with PBST: 444.4nmol / ml, 222.2nmol / ml, 111.1nmol / ml, 55.6nmol / ml, 27.8nmol / ml, 0nmol / ml; operation process: equilibrate in buffer 1 (PBST) for 60s, immobilize antibody in antibody solution for 300s, incubate in buffer 2 (PBST) for 180s, bind in antigen solution for 420s, dissociate in buffer 2 for 1200s, use 10mM The pH 1.69 GLY solution and buffer 3 were used to regenerate the sensor and output data. The results are shown in Table 1. (KD stands for equilibrium dissociation constant or affinity)

[0043] Table 1 Affinity results of 2F6 monoclonal antibody

[0044] Antibody name KD(M) 2F6 monoclonal antibody 9.86E-09

[0045] It can be seen from the results in Table 1 that the 2F6 monoclonal antibody...

Embodiment 3

[0054] Example 3 Cultivation of fibroblasts

[0055] The fibroblast HSF was cultured in DMEM (containing 10% fetal bovine serum, 1% double antibody mixture) medium, placed in a 37°C, 5% CO2 incubator, and the cells grew to more than 80%, rinsed with PBS for 2- 3 times, add 1 mL of 0.25% trypsin to digest at room temperature for 30-60 s, tap the bottle wall to make the cells fall off, add 2 times the volume of fetal bovine serum-containing medium to terminate the digestion, pipette evenly with a gun tube, and centrifuge at 1000 r / min for 8 min , remove the supernatant, precipitate and add an appropriate amount of complete medium to make a single cell suspension, adjust the concentration, and use the obtained HSF in logarithmic growth phase in good condition for the experiment. HSF in logarithmic growth phase was inoculated into a 96-well culture plate at 1×105 cells / mL, and 90 μL was inoculated in each well, placed in a cell incubator at 37 °C, 5% CO2, and taken out after 24 h ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
diameteraaaaaaaaaa
Login to View More

Abstract

The invention relates to a medicine or a cosmetic prepared from fibroblast extracellular vesicles. The effect of the invention is achieved by inhibiting P53 activity to improve fibroblast activity and prevent aging to produce ectovesicles at high yield. A monoclonal antibody is used for inhibiting the activity of P53, after the monoclonal antibody is added into a culture medium of fibroblasts, the aging of the fibroblasts can be inhibited, the content of exovesicles secreted by the fibroblasts can be increased, and the exovesicles are good in anti-aging activity and suitable for preparing anti-aging beautifying or medicine products. Good market application values are realized.

Description

technical field [0001] The invention relates to the field of pharmacy, in particular, to medicines or cosmetics prepared from fibroblast extracellular vesicles. Background technique [0002] External vesicles, also called exosomes, are double-layered phospholipid vesicles with a diameter of 30-150 nm secreted by cells, which contain a variety of miRNAs, proteins and cellular regulatory factors; studies have found that exosomes can communicate between cells. , transmit biological information and proteins, and play a role in regulating receptor cells. [0003] Extracellular vesicles are widespread in the human body and play an important role in physiological and pathophysiological processes. Extracellular vesicles have been identified in a variety of body fluids, which is the basis for their use as disease markers, so scientists and clinicians are actively studying their role in diagnosis. The first evidence for the interaction of extracellular vesicles with recipient cells ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18C12N15/13C12N5/077A61K8/99A61K39/395A61K48/00A61P17/18
CPCC07K16/18C12N5/0656A61K48/005A61K39/39533A61K8/99A61P17/18C07K2317/565C07K2317/56C12N2509/00A61K2800/86
Inventor 李少波亓爱杰黄昱李立华陈清轩
Owner 诺赛联合(北京)生物医学科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com