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Production method of complete peptide chain bovine collagen

A collagen and production method technology, applied in the field of medical biomaterials, can solve the problems that collagen does not have peptide chains, is expensive, and has toxic substances in collagen, and achieves simple production process, short enzymatic hydrolysis time, and biophase capacitive ideal effect

Inactive Publication Date: 2019-02-26
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, collagen extraction mainly comes from animal bones, but animal bones contain a large amount of inorganic substances, so the extraction of collagen from animal bones is cumbersome and the yield is low, especially the collagen obtained by high temperature combined with enzyme treatment. The peptide chain will be degraded to varying degrees, that is, the obtained collagen does not have a complete peptide chain; the existing technology uses alkali extraction method to easily cause collagen denaturation, and severe hydrolysis may even lead to racemization of collagen and produce toxic substances
In addition, the commercialized genetically recombinant collagen is very expensive, the price is about 100,000 RMB per gram, and the peptide chain of this recombinant collagen also degrades to varying degrees, so it can only be used to coat cell growth medium, thereby promoting the attachment of cells, and these collagens with incomplete peptide chains cannot meet the needs of the practical application of collagen in medical biomaterials, especially in the preparation of tissue engineering carrier scaffolds, let alone large-scale Production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] First remove the fascia, fat, and blood vessels from the fresh beef Achilles tendon, then clean it, drain the water, and cut it into small pieces;

[0041] Take 900 mL of distilled water, add 29.25 g of NaCl and 6.057 g of Tris-HCl, after completely dissolving, adjust the pH to 7.4, and dilute to 1000 mL with distilled water to prepare a non-collagen protein removal solution; take 0.3 g of freeze-dried bovine Achilles tendon , soak in 900 mL of non-collagen protein removal solution and stir continuously for 48 h, replace the solution every 12 h, then discard the supernatant, wash with distilled water until the pH is neutral, and drain;

[0042] Then take 4800 mL of distilled water, add 90 mL of glacial acetic acid, and then distill the water to 5000 mL to make a collagen solution; add 300 mL of collagen solution and stir for 24 h;

[0043] Put the above mixed solution into a homogenizer to fully homogenize, then add 600 mL of collagen solution, and stir for 48 h;

[00...

Embodiment 2

[0051] First remove the fascia, fat, and blood vessels from the fresh beef Achilles tendon, then clean it, drain the water, and cut it into small pieces;

[0052] Take 900 mL of distilled water, add 58.5 g of NaCl and 6.057 g of Tris-HCl, after completely dissolving, adjust the pH to 7.4, and dilute to 1000 mL with distilled water to prepare a non-collagen protein removal solution; take 0.3 g of freeze-dried bovine Achilles tendon , soak in 900 mL non-collagen protein removal solution and stir continuously for 36 hours, replace the solution every 9 hours, then discard the supernatant, wash with distilled water until the pH is neutral, and drain;

[0053] Then take 4800 mL of distilled water, add 120 mL of glacial acetic acid, and then distilled water to 5000 mL to prepare a collagen solution; add 300 mL of collagen solution, and stir for 18 h;

[0054] Then put the above mixed solution into a homogenizer to fully homogenize, then add 600 mL of collagen solution, and stir for 3...

Embodiment 3

[0062] First remove the fascia, fat, and blood vessels from the fresh beef Achilles tendon, then clean it, drain the water, and cut it into small pieces;

[0063] Take 900 mL of distilled water, add 117 g of NaCl and 6.057 g of Tris-HCl, after completely dissolving, adjust the pH to 7.4, and dilute to 1000 mL with distilled water to prepare a non-collagen protein removal solution; take 0.3 g of freeze-dried bovine Achilles tendon , soak in 900 mL non-collagen protein removal solution and stir continuously for 24 h, replace the solution every 8 h, then discard the supernatant, wash with distilled water until the pH is neutral, and drain;

[0064] Then take 4800 mL of distilled water, add 180 mL of glacial acetic acid, and then distilled water to 5000 mL to prepare a collagen solution; add 300 mL of collagen solution, and stir for 12 h;

[0065] Put the above mixed solution into a homogenizer to fully homogenize, then add 600 mL of collagen solution, and stir for 24 h;

[0066]...

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Abstract

The invention discloses a production method of a complete peptide chain bovine collagen. The production method is characterized in that the operations are conducted under a low temperature condition after a bovine achilles tendon is pretreated, and the method comprises the following steps: firstly processing by using a non-collagen removing solution, then dissolving by using a collagen dissolvingsolution, supplementing the collagen dissolving solution to continue to dissolve the collagen after fully homogenating, then adding collagen enzymolysis liquid to implement enzymolysis, collecting supernate after centrifuging, then adding a collagen precipitation solution to precipitate the collagen, collecting a precipitate after centrifuging, repeating the processing steps of dissolving the collagen by using the collagen dissolving solution, centrifuging to collect the supernate, precipitating the collagen by using the collagen precipitation solution, centrifuging to collect the precipitate,dissolving by using the collagen dissolving solution and centrifuging to collect the supernate, finally implementing dialysis on the supernate to obtain the complete peptide chain bovine collagen, and storing after freeze-drying. The complete peptide chain bovine collagen can be applied to the preparation of tissue engineering carrier supports, surgical sutures, wound-protecting dressings and thelike as a medical biological material.

Description

technical field [0001] The invention belongs to medical biomaterials, in particular to a method for producing complete peptide chain bovine collagen. Background technique [0002] As a natural biomacromolecule, collagen has been widely used in food, biomedicine and cosmetics because of its good biocompatibility, degradability and biological activities such as wound repair, hemostasis, anti-radiation and anti-aging. and other fields. At present, collagen extraction mainly comes from animal bones, but animal bones contain a large amount of inorganic substances, so the extraction of collagen from animal bones is cumbersome and the yield is low, especially the collagen obtained by high temperature combined with enzyme treatment. The peptide chain will be degraded to varying degrees, that is, the obtained collagen does not have a complete peptide chain; the existing technology uses alkali extraction method to easily cause collagen denaturation, and severe hydrolysis may even lea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/78C07K1/14C12P21/06A61L27/24A61L17/10A61L15/32
CPCA61L15/325A61L17/10A61L27/24A61L2430/16C07K14/78C12P21/06
Inventor 徐彬樊廷俊姜国建
Owner OCEAN UNIV OF CHINA
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