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Double function epidermal growth factor and its preparation method and uses

An epidermal growth factor, dual-function technology, used in skin care preparations, chemical instruments and methods, skin diseases, etc., can solve the problem of non-specificity of damaged tissue, lack of specificity of wound tissue, and repair performance of gastrointestinal ulcers Poor and other problems, to achieve the effect of clear drug efficacy, no adverse side effects, and low cost

Inactive Publication Date: 2006-11-22
北京上地新世纪生物医药研究所有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the recombinant EGF encounters the following difficulties in clinical practice: ① EGF has no specificity to damaged tissues, so it has unsatisfactory effects on wound healing such as burns and wounds; Worse, mainly due to the lack of specificity combined with wound tissue; ③ The half-life of EGF in the body is very short, especially when applied to gastrointestinal ulcers, free EGF is easily hydrolyzed by proteolytic enzymes of cells

Method used

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  • Double function epidermal growth factor and its preparation method and uses
  • Double function epidermal growth factor and its preparation method and uses
  • Double function epidermal growth factor and its preparation method and uses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1: Construction of PSL-CBD-EGF recombinant plasmid

[0025] 1. Using the primers P1 and P2 shown in Figure-1, the cDNA of L is used as a template to synthesize EGF PCR using high-fidelity TaqPCR enzyme.

[0026] serial number

name

Specification

unit

quantity

1

10× buffer

microliter

10

2

magnesium sulfate

50mmol / L

microliter

4

3

P1 primer

5mm

microliter

2

4

P2 primer

5mm

microliter

2

5

cDNA

100ng

microliter

1

6

Taq enzyme

5u / ul

microliter

1

7

sterile water

microliter

72

8

DNTP(10mm)

microliter

8

[0027] After preparing the above reaction system on an ice bath, centrifuge for 1 minute (3000 rpm) to mix the components in the system, and then place the sample in a PCR synthesizer. The conditions for the synthesis reaction a...

Embodiment 2

[0077] Example 2: Evaluation of CBD binding activity in EGF-CBD

[0078] In the 4 sets of experiments in Figure-3, the petri dishes were all coated with collagen, and then the test reagents were added:

[0079] A. Medium

[0080] B. Medium + CBD

[0081] C. Medium + EGF

[0082] D. Medium + EGF-CBD

[0083] Incubate at 37°C for 24 hours, then discard the culture medium in each plate, wash the culture plate with buffer solution (PBS) four times, and then detect with EGF antibody, the results show that: in group D, the CBD in EGF-CBD can Effectively bind to collagen molecules. At the same time, culture "NRK49F" cells with A.B.C.D four-group test-treated dishes. After inoculating the same number of cells, culture them at 37°C for 48 hours. Microscopic examination shows that EGF-CBD can Very effective in promoting cell growth.

Embodiment 3

[0084] Example 3: Comparison of EGF-CBD and EGF activity

[0085] Group 1: Medium

[0086] Group 2: Medium + CBD

[0087] Group 3: Medium + EGF

[0088] Group 4: culture medium + EGF-CBD

[0089] Figure-4 shows that each group was inoculated with the same number of cells (NRK49F). After culturing at 37°C for 48 hours, the growth rate of the cells is: EGF-CBD>EGF>CBD≥Medium, which shows that EGF-CBD can effectively promote cell growth like EGF.

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Abstract

The invention discloses a human epiderm growth factor with 53 amino acids to form cellar factor, which is characterized by the following: forming molecular bridge of four amino acids to connect human epiderm growth factor and collagen protein combination region; producing the fusion protein with high biological activity and high specific collagen combination activity. The double-functional epiderm growth factor can restore human internal and external wound, which can be applied in the cosmetics.

Description

technical field [0001] The invention relates to human epidermal growth factor in genetic engineering in the field of biochemical technology. Specifically, a fusion protein with high biological activity and high specific collagen binding activity—bifunctional epidermal growth factor is obtained through gene recombination technology. Background technique [0002] Epidermal Growth Factor (EGF) was first discovered by American scientist Cohen in 1962. Cohen found that there is a polypeptide composed of 53 amino acids in the submandibular gland of mice, which can widely promote cell proliferation, and named it epidermal growth factor Growth factor (EGF), in 1972, analyzed and determined the amino acid composition and structural characteristics of mouse EGF. In 1975, human EGF was extracted from urine for the first time, and it was found that it can inhibit the secretion of gastric acid, so it is also called gastrin. It is now clear that EGF is a class of polypeptides widely pres...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00A61K8/64A61K38/18A61P17/02A61Q19/00
Inventor 隋德新李晶
Owner 北京上地新世纪生物医药研究所有限公司
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