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Preparation process for collagen sponge

A collagen sponge and pigskin technology, applied in the field of protein engineering, can solve the problems of inability to retain active collagen triple helix structure, low extraction efficiency and purity of collagen, and destruction of triple helix structure, and achieve low immunogenicity and high biological efficiency. Compatibility, the effect of improving extraction efficiency

Active Publication Date: 2013-03-13
无锡贝迪生物工程股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Disclosed in CN101005865A is a kind of preparation method of collagen protein sponge, carries out cross-linking with glutaraldehyde, and its collagen extraction efficiency and purity are all low, and cross-linking agent uses glutaraldehyde safety lower
The methods disclosed in the prior art often destroy the triple helix structure of collagen during the extraction and preparation of collagen, or the extraction efficiency is low, and it is impossible to preserve the triple helix structure of active collagen while improving the extraction efficiency and purity. Preparation of safe collagen sponge by enzymatic cross-linking of collagen

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] (1) Take fresh cowhide, cut and remove the grease layer and fur layer, remove impurities, wash, break the cowhide into small particles, add 0.03mol / L sodium hydroxide aqueous solution according to the ratio of solid to liquid ratio of 1:30, and Soak at 6-8°C for 2 hours, filter, and set aside;

[0027] (2) Add anhydrous ether or acetone with a quality 8 times that of the cowhide to the cowhide after the above pretreatment, reflux at 35°C for 8 hours, then rinse with distilled water until there is no peculiar smell, and set aside;

[0028] (3) immerse the above-mentioned cowhide in the mixed solution of 0.6mol / L glacial acetic acid and 700mg / L pepsin, and keep stirring for about 25 hours;

[0029] (4) Adopt progressive ultrasonic treatment to the above mixture, first use 100W ultrasonic treatment for 30 minutes, then use 300W ultrasonic treatment for 30 minutes, and finally use 400W ultrasonic treatment for 1 hour;

[0030] (5) Add 2% H 2 o 2 Solution, mix and stand f...

Embodiment 2

[0036] (1) Take fresh beef Achilles tendon, cut and remove the grease layer and fur layer, remove impurities, wash, break the beef Achilles tendon into small particles, and add 0.01mol / L of hydrogen peroxide according to the solid-liquid ratio of 1:40 Sodium aqueous solution, soak at 6-8°C for 1 hour, filter, and set aside;

[0037] (2) Add anhydrous ether or acetone with a mass 6 times the mass of the Achilles tendon to the Achilles tendon after the above pretreatment, reflux at 40°C for 5 hours, then rinse with distilled water until there is no peculiar smell, and set aside;

[0038] (3) immerse the above-mentioned bovine Achilles tendon in the mixed solution of 0.7mol / L glacial acetic acid and 600mg / L pepsin, and keep stirring for about 30 hours;

[0039] (4) Adopt progressive ultrasonic treatment to the above mixture, first use 100W ultrasonic treatment for 30 minutes, then use 200W ultrasonic treatment for 30 minutes, and finally use 300W ultrasonic treatment for 1 hour;

...

Embodiment 3

[0046] (1) Take fresh pigskin, cut and remove the grease layer and fur layer, remove impurities, wash, break the pigskin into small particles, and add 0.02mol / L sodium hydroxide aqueous solution according to the ratio of solid to liquid 1:35 , soaked at 6-8°C for 1.5 hours, filtered, and set aside;

[0047] (2) Add anhydrous ether or acetone whose mass is 7 times that of the pigskin to the pigskin after the above pretreatment, reflux at 37° C. for 6 hours, then rinse with distilled water until there is no peculiar smell, and set aside;

[0048] (3) immerse the above-mentioned pigskin in the mixed solution of 0.65mol / L glacial acetic acid and 650mg / L pepsin, and keep stirring for about 28 hours;

[0049] (4) Adopt progressive ultrasonic treatment to the above mixture, first use 150W ultrasonic treatment for 30 minutes, then use 250W ultrasonic treatment for 30 minutes, and finally use 350W ultrasonic treatment for 1 hour;

[0050] (5) Add 2.5% H 2 o 2 solution, mix and stand...

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Abstract

The invention relates to a preparation method for collagen sponge. The preparation method comprises the following steps of: pretreating fresh pig skin serving as a raw material to remove fat; treating by an acid enzyme method; performing ultrasonic treatment so as to further improve extraction efficiency; purifying to prepare a high-purity collagen product; and preparing the collagen sponge through enzyme method crosslinking and freeze drying. By the preparation method for the collagen sponge, the extraction efficiency is high and the product purity reaches over 98 percent. The product comprises the collagen sponge which is prepared by freeze-drying active collagen and maintains a triple helix structure, has low immunogenicity, high biocompatibility and biodegradability, and is applicable to medical treatment.

Description

technical field [0001] The invention relates to a preparation method of a collagen sponge, which belongs to the field of protein engineering. The product is prepared by cross-linking active collagen maintaining a triple helix structure, has low immunogenicity, high biocompatibility, is biodegradable, and is suitable for for medical use. Background technique [0002] Collagen is a group of hard proteins with the most abundant content and widest distribution in vertebrates. 25%-33% of the total protein content of the animal body. Several types of collagen are often contained in the same tissue, and a certain type is often the main one. Type I collagen is distributed in all parts of the body, mainly in the skin, tendons and ligaments, and has strong tensile strength. Type II collagen mainly exists in hyaline cartilage and vitreous body, and has a strong ability to resist pressure. Type III collagen is widely distributed in tissues with high extensibility, such as loose conn...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/02C07K14/78C07K1/36
Inventor 任伟业
Owner 无锡贝迪生物工程股份有限公司
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