Soluble human keratin and application thereof

A soluble and keratin technology, applied in the field of biomedicine, can solve the problems of keratin natural structure damage, impact on stability and safety, use restrictions, etc., to avoid rejection, the preparation method is green and mild, keratinocyte proliferation and The effect of migration promotion

Active Publication Date: 2019-08-13
海默斯(重庆)医学生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The natural structure of keratin after acid-base and redox treatment is destroyed, affecting its stability and safety
And because the source of keratin is wool, it may cause rejection when it is used for medical purposes, so its use has certain restrictions

Method used

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  • Soluble human keratin and application thereof
  • Soluble human keratin and application thereof
  • Soluble human keratin and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] The preparation of embodiment 1 soluble keratin

[0064] 1. Amplify the target fragment

[0065] 1) Synthesis of the target gene

[0066] According to the nucleotide sequence of the target gene shown, Yu Sangon Bioengineering (Shanghai) Co., Ltd. carried out whole gene synthesis and sequencing verification to obtain a template gene, as shown in SEQ ID NO:1.

[0067] 2) According to the nucleotide sequence of the target gene, respectively design primers, as shown in SEQ ID NO: 4-7. The synthesized template gene was used as a template, and the PCR amplification was carried out with PRO-1-F and PRO-1-R as primers.

[0068] PCR reaction system: 1 μL of 10 μmol / L primer, 1 μL of target gene or linearized pET28a-His-TEV vector gene, 4 μL of dNTP (each 2.5 mM), 10×Buffer (containing Mg 2+ ) 5 μl, 1 μL Pfu DNA Polymerase, add water to make up to a total volume of 50 μL.

[0069] PCR reaction conditions: pre-denaturation at 95°C for 5min; denaturation at 98°C for 10s, anneal...

Embodiment 2

[0089] The characteristic of embodiment 2 target protein

[0090] 1. Swelling test

[0091] The soluble human keratin of the present invention has the characteristic of rapid swelling, and the complete swelling time of the soluble human keratin of the present invention is about 2 hours. After the protein of the present invention is swollen, a viscous and clear liquid is obtained, and the viscosity is about 5000 mPa.s at a concentration of 2%.

[0092] The swelling test shows that the keratin obtained by the present invention is soluble, and the safe soluble keratin can be obtained without chemical treatment.

[0093] 2. Effect on cell proliferation

[0094] 1) The soluble human keratin of the present invention is formulated with deionized water to a concentration of 5 mg / ml (solution A), fully swelled by shaking for 24 hours, and solution A is diluted to 2.5 mg / ml with deionized water respectively (solution B). ), 1.25 mg / ml (solution C), 0.625 mg / ml (solution D), 0.313 mg / ...

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Abstract

The invention belongs to the field of biomedicine and particularly relates to soluble human keratin and application thereof. The soluble keratin is obtained through the following process of cloning amiddle sequence of a human gene K37, constructing a recombinant vector, transforming bacteria, inducing expression of the recombinant protein and purifying the recombinant protein to obtain the soluble keratin. The soluble keratin has the characteristic of rapid swelling and can promote mitosis and migration of epidermal cells, and therefore the soluble keratin can be used for preparing biologicalmaterials or medicaments for promoting cell proliferation or migration, and plays a role in hemostasis and wound healing. Moreover, the soluble keratin causes no rejection phenomenon when being usedfor the human body, is safe to use and thus has a wide application prospect in the field of biomedicine.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a preparation method of soluble human keratin and its application in the field of biomedicine. Background technique [0002] Keratin is a kind of fibrous animal protein with connective and protective functions. It is a structural protein of ectoderm cells and widely exists in animal hair, feathers and hooves. All keratins have the characteristic of homocysteine, which forms disulfide bonds between keratins, which provide keratin with the required mechanical properties, and also cause keratin insolubility, which facilitates the utilization of keratin bring obstacles. Keratin has good bioactivity and tissue compatibility, can play the role of anti-inflammatory, hemostasis, nerve repair, reduce wound exudation and promote wound tissue regeneration, repair and healing, and can be biodegraded in the body, so it can be used as a good biomedical materials. [0003] At present, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C12N15/12C12N5/071C12N15/70C12N1/21A61K38/17A61P7/04A61P17/02C12R1/19
CPCC07K14/4741C12N5/0629C12N15/70A61P7/04A61P17/02C12N2501/998A61K38/00
Inventor 王伯初郝石磊管璐晗
Owner 海默斯(重庆)医学生物技术有限公司
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