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Method for preparing medical III type collagen

A collagen and nascent technology, applied in the field of preparation of medical grade III collagen, can solve the problems of complicated operation, many limiting factors, low yield, etc., and achieve the effects of abundant sources, scientific and advanced preparation methods, and high product recovery rate.

Inactive Publication Date: 2012-07-04
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

High performance liquid phase ion exchange column chromatography, CM-cellulose column chromatography and agarose molecular sieve gel filtration column chromatography have a high degree of purification, but the yield is low and the operation is complicated, so there are more limiting factors

Method used

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  • Method for preparing medical III type collagen
  • Method for preparing medical III type collagen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Preparation method of medical grade type III collagen

[0029] 1. Pretreatment of newborn calves

[0030] Shave off the hair and hair follicles of fresh newborn calves, remove the subcutaneous tissues and fat, and then finely cut them. Take 200g of the finely sliced ​​skin and add 2000ml of a mixed solution of alcohol: haloalkane = 0.5 by volume, soak it for degreasing, and replace the above solution every 5 hours , Pour out the solution in 15 hours; soak in 2000ml of 98% ethanol aqueous solution, change once in 5 hours, and pour out the concentration; wash once with 2000ml of 20% ethanol aqueous solution; then use 0.1mol / L EDTA solution, pH=10.5~11.5, soak at 4℃ for 6 hours, then wash 3 times with 2000ml of ultrapure water; then soak with 2000ml of 10% NaCl aqueous solution, change once every 4 hours, pour out the solution and use ultrapure water Wash twice with 2000ml of pure water, then put the cowhide in 2000ml of ultrapure water to soak overnight, and pour ou...

Embodiment 2

[0038] Example 2 Preparation method of medical grade type III collagen

[0039] 1. Pretreatment of newborn calves

[0040] The fresh newborn calves are shaved off the hair and follicles, and the subcutaneous tissues and fat are removed and finely cut. Take 300g of finely sliced ​​skins and add 5000ml of mixed solution of alcohol: haloalkane = 1.5 volume ratio, soak for degreasing, replace the above solution every 15 hours, pour out the solution; soak in 5000ml of 98% ethanol aqueous solution for 15 hours Replace it once, pour out the concentration in 30 hours; wash once with 5000ml of 20% ethanol aqueous solution; then use 0.2mol / L EDTA solution, pH=10.5~11.5, soak at 6℃ for 12 hours, then use ultrapure water Wash 3 times with 5000ml; then soak with 5000ml of 30% NaCl aqueous solution, change once every 6 hours, pour out the solution, wash 2 times with 5000ml ultrapure water, and then put the cowhide into 5000ml ultrapure water to soak overnight. Pour out the water.

[0041] 2. Pe...

Embodiment 3II

[0048] Example 3 Preparation method of type III collagen

[0049] 1. Pretreatment of newborn calves

[0050] The fresh newborn calves are shaved off the hair and follicles, and the subcutaneous tissues and fat are removed and finely cut. Take 250g of shredded skin and add 3000ml of mixed solution of alcohol: haloalkane = 1 volume ratio, soak for degreasing, replace the above solution every 10 hours, pour out the solution; soak in 3000ml of 98% ethanol aqueous solution for 10 hours Replace it once, discard the concentration in 20 hours; wash once with 3000ml of 20% ethanol aqueous solution; then use 0.15mol / L EDTA solution, pH=10.5~11.5, soak at 4℃ for 9 hours, then use ultrapure water Wash 3000ml for 3 times; then soak it with 3000ml of 20% NaCl aqueous solution, change it every 5 hours, pour out the solution, wash twice with 3000ml ultrapure water, and soak the cowhide in 3000ml ultrapure water overnight. Pour out the water.

[0051] 2. Pepsin digestion

[0052] In the above solid...

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Abstract

The invention discloses a method for preparing a medical III type collagen, which is characterized by comprising the following steps of: pre-treating fresh newborn kip as a raw material and then extracting a mixture of I type and II type collagens by using a pepsin degradation limiting method and a salting out method; dissolving the mixture with 1.5 mol / L NaCl and 0.05mol / L Tris buffer solution the pH value of which is 7.5 for 2-5 times to remove most of I type collagen; heating the mixture with 2-6mol / L guanidine hydrochloride and 0.05mol / L Tris-HCl buffer solution the pH value of which is 7.5 in a water bath with the temperature of 45-95 DEG C for 30-60 minutes to modify the collagens; reducing the temperature and the concentration of the guanidine hydrochloride; renaturing the II type collagen after 2-6 hours; and centrifuging the mixture at high speed to obtain a III type collagen.

Description

Technical field [0001] The invention relates to a method for preparing medical grade type III collagen, which belongs to the field of preparation of biomedical materials. Background technique [0002] Type III Collagen (CIII) is a typical interstitial collagen, which is mainly found in tissues such as skin, blood vessels, heart, lung, liver and kidney. Type III collagen in the form of fine fibers endows the tissue with a strong ability to expand, and plays an important role in maintaining the elasticity of skin, blood vessels, liver and other tissues and organs, and maintaining their normal physiological functions. Recent studies have shown that type III collagen plays an important role in the pathological study of Ehlers-Danlos syndrome IV (EDS IV) and aneurysms (Kroes HY, Pals G and van Essen AJ. Ehlers-Danlos syndrome type IV: unusual congenital anomalies in a mother and son with a COL3A1 mutation and a normalcollagen III protein profile.Clin Genet.2003:63:224-227 and Brega K...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/06C07K14/78A61K38/39A61L27/24
Inventor 蒋波李霞陈露万法盛
Owner SICHUAN UNIV
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