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Method for rapidly extracting high molecular weight silk fibroin

A silk fibroin, high molecular weight technology, applied in the field of natural polymers, can solve the problems of lack of processability and insufficient scaffold materials, and achieve the effects of good biocompatibility, short production cycle and high biosafety

Inactive Publication Date: 2014-11-26
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method only briefly describes how to extract silk glands from the middle silk gland, and the silk fibroin obtained by this method is actually a silk gel rather than a silk solution
Silk fibroin in the gel state can be directly dried to obtain silk fibroin powder. However, silk fibroin gels are far from enough to prepare scaffold materials with high strength mechanical properties and regular shapes, so they do not have processability.

Method used

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  • Method for rapidly extracting high molecular weight silk fibroin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The method for extracting silk fibroin in this embodiment includes the following steps.

[0025] (1) Pull out the silk gland of the five-year-old silkworm with medical forceps, remove the epithelial tissue, and obtain a strip of protein;

[0026] (2) soaking the protein in step (1) in deionized water and rinsing to remove sericin, thereby obtaining silk fibroin;

[0027] (3) Place the silk fibroin in step (2) in 0.5wt% sodium carbonate aqueous solution, shake and dissolve at a rotating speed of 50rpm, and obtain a silk fibroin solution with a mass fraction of 20%. The electrophoresis results show that the obtained The molecular weight of silk fibroin is higher than 250KDa.

Embodiment 2

[0029] The method for extracting silk fibroin in this embodiment includes the following steps.

[0030] (1) Use medical forceps to pull out the silk glands of the fifth instar silkworm, remove the epithelial tissue of the front, back and middle silk glands, and obtain a section of clear and transparent strip-like protein, which is is silk gland protein;

[0031] (2) immersing the silk adenin in the step (1) in deionized water and rinsing to remove the sericin, thereby obtaining a silk fibroin gel;

[0032] (3) the silk fibroin in the step (2) is placed on the gel to collect 0.2wt%Na 2 HPO 4 In the buffer solution, place on a shaker and shake at a speed of 200 rpm for 10 minutes, then shake at a speed of 50 rpm for 1 hour to obtain a solution;

[0033] (4) Adding the silk fibroin solution in step (3) to the cell culture medium for cell culture can significantly promote the adhesion and proliferation of human MSC cells.

Embodiment 3

[0035] The method for extracting silk fibroin in this embodiment includes the following steps.

[0036] (1) Use medical forceps to pull out the silk glands of the fifth instar silkworm, remove the epithelial tissue of the front, back and middle silk glands, and obtain a section of clear and transparent strip-like protein, which is is silk gland protein;

[0037] (2) immersing the silk adenin in the step (1) in deionized water and rinsing to remove the sericin, thereby obtaining a silk fibroin gel;

[0038] (3) the silk fibroin in the step (2) is placed on the gel to collect 0.2wt%Na 2 HPO 4 In the buffer solution, place on a shaker and shake at a speed of 200 rpm for 10 minutes, then shake at a speed of 50 rpm for 1 hour to obtain a solution;

[0039] (4) Adding the silk fibroin solution in step (3) to the cell culture medium for cell culture can significantly promote the adhesion and proliferation of MC-3T3-E1 cells.

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Abstract

The invention discloses a method for rapidly extracting high molecular weight silk fibroin, which is characterized in that a pair of silk gland is removed from maturated insect, epithelial tissue on outer layer is removed to obtain a segment of clear and transparent band, the band protein is immersed in deionized water or PBS for rinsing and removing sericin, and thereby the silk fibroin is obtained. The method has the advantages of simple operation, short production period, simple extraction process and high efficiency, complex and tedious material extraction process is not required, and time is greatly saved. In the method, no other toxic chemical reagent is used during the extraction process, the preparation condition is mild, cost is greatly reduced, other waste is not generated, and zero pollution is generated for environment.

Description

technical field [0001] The invention relates to a method for extracting silk fibroin solution and silk powder, and belongs to the field of natural polymers. Background technique [0002] Silk proteins derived from silkworms such as silkworms, wild silkworms (such as tussah silkworms, celestial silkworms, and castor silkworms) and spiders have excellent biocompatibility, no immune rejection, biodegradability, and are essentially natural proteins. , cosmetics, food, environmental protection, biomaterials, tissue engineering and other fields have received high attention. Natural silk protein is synthesized and secreted in insects, and is stored in various silk glands in the form of silk protein solution with high concentration (the silk gland protein in the middle part of silkworm can reach 30wt%). [0003] However, when insects spit out silk protein, it immediately solidifies into silk fibers. At this time, it is difficult to dissolve silk fibers in common buffer solvents, ma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C07K1/14
CPCC07K14/43586C07K14/43518
Inventor 杨明英帅亚俊
Owner ZHEJIANG UNIV
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