Eureka AIR delivers breakthrough ideas for toughest innovation challenges, trusted by R&D personnel around the world.

Transforming growth factor beta1 active polypeptide and application thereof

A transforming growth factor and active polypeptide technology, applied in the direction of transforming growth factor, growth factor/inducing factor, specific peptide, etc., can solve the problem that there is no transforming growth factor β1 active polypeptide gel, etc., to promote wound healing, improve Healing efficiency and effectiveness, inflammation-reducing effect

Pending Publication Date: 2022-04-29
PLASTIC SURGERY HOSPITAL CHINESE ACAD OF MEDICAL SCI
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But at present, there is no relevant report on the use of transforming growth factor β1 in the preparation of active polypeptide gels.

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
  • Transforming growth factor beta1 active polypeptide and application thereof
  • Transforming growth factor beta1 active polypeptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1: Screening of TGF-β1 active polypeptide sequence

[0032] Coat the 96-well plate with human TGF-β1 monoclonal antibody, add phage random dodecapeptide library (purchased from NEB Company, USA) and incubate overnight at 4°C, wash the plate 10 times with TBST, and wash the plate with elution buffer (0.2mol / L Glycine-HCl, pH 2.2, 0.1% BSA) to elute the bound phages, neutralization buffer (1mol / L Tris-HCl, pH 9.1) quickly neutralizes the eluate.

[0033] E.coli ER2738 host bacteria were used to amplify the eluate for the next round of screening. In the same way, three rounds of screening were performed on the eluate from each round. Titer determination was carried out on the eluate of the fourth round, phage coeruleus was randomly selected, amplified, and adjusted to 2.0×10 13 pfu / mL.

[0034] The ELISA method and the human TGF-β1 immunoassay kit (purchased from R&D Company, USA) were used to detect the binding ability of the monoclonal phage according to th...

Embodiment 2

[0044] Embodiment 2: Synthesis of TGF-β1 active polypeptide

[0045] The seven TGF-β1 polypeptide sequences screened in Example 1 were sent to Wuhan Minghao Biotechnology Co., Ltd. for the synthesis of TGF-β1 active polypeptides, purified to a purity of 98%, and then disulfide bonds were used to stabilize the peptides. Chain structure, at the same time, the C-terminus of the polypeptide is amidated and blocked to enhance the stability of the active polypeptide; the N-terminus of the polypeptide is labeled with a rhodamine laser dye to identify the affinity of the active polypeptide with fibroblasts, and to detect whether it can Associated with fibroblasts. Prepared into freeze-dried powder and stored at -20°C.

Embodiment 3

[0046] Embodiment 3: In vivo and in vitro experiments of TGF-β1 active polypeptide

[0047] The TGF-β1 active polypeptides in Example 2 are intervened in the cultured fibroblasts, because the TGF-β1 active polypeptides are labeled with rhodamine laser dye, so immunofluorescence detection can be used to detect their affinity with fibroblasts . The results show that only the TGF-β1 active polypeptide shown in SEQ ID NO.23 can bind to the specific receptor on fibroblasts.

[0048] Further, the TGF-β1 active polypeptide shown in SEQ ID NO.23 was co-cultured with human fibroblasts for in vitro experiments, and a blank control group, a TGF-β1 active polypeptide group and a TGF-β1 growth factor group were set. Using the CCK-8 method, it was found that compared with the negative control group, the TGF-β1 active polypeptide group could promote cell proliferation, and there was a significant difference (P<0.05), and it was dose-dependent. See Table 3 for details.

[0049] Table 3 CCK-...

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

No PUM Login to View More

Abstract

The invention discloses a transforming growth factor beta 1 active polypeptide and application thereof, and belongs to the technical field of transforming growth factor beta 1 active polypeptides. The amino acid sequence of the transforming growth factor beta 1 active polypeptide is as shown in SEQ ID NO. 23. The invention also discloses application of the transforming growth factor beta1 active polypeptide in preparation of drugs for wound healing. According to the invention, a phage random dodecapeptide library is used for screening a TGF-beta1 key sequence, in-vitro synthesis of the TGF-beta1 active polypeptide is carried out, a large amount of TGF-beta1 active polypeptide can be synthesized in a short time, and the production cost is greatly reduced.

Description

technical field [0001] The invention relates to a transforming growth factor β1 active polypeptide and application thereof, belonging to the technical field of transforming growth factor β1 active polypeptide. Background technique [0002] Wound healing is a complex process of tissue defect repair that requires the coordinated and orderly participation of various cells, extracellular matrix and signaling pathways. At present, refractory wounds such as diabetic chronic wounds are clinical problems that need to be solved urgently. Topical application of growth factors can promote wound healing, but currently used growth factor products have complex production processes, high economic costs, short biological half-life, poor stability, and are difficult to achieve the expected therapeutic effect, which greatly limits their clinical application. [0003] At present, the research and application of peptides have reached an unprecedented level of prosperity. Compared with natural...

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
IPC IPC(8): C07K14/495A61K38/18A61K9/06A61K47/32A61K47/10A61K47/14A61K47/36A61P17/02A61P29/00
CPCC07K14/495A61K9/06A61K9/0014A61K47/32A61K47/10A61K47/14A61K47/36A61P17/02A61P29/00A61K38/00
Inventor 宗宪磊杜宏靳小雷宋国栋
Owner PLASTIC SURGERY HOSPITAL CHINESE ACAD OF MEDICAL SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
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