Method for preparing medical III type collagen

A collagen and raw material technology, applied in the field of preparation of medical grade III collagen, can solve the problems of many limiting factors, complicated operation, low yield, etc., and achieve the effect of abundant sources, simple process and high product recovery rate

Inactive Publication Date: 2010-06-02
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

High performance liquid phase ion exchange column chromatography, CM-cellulose column chromatography and agarose molecular sieve gel filtratio...

Method used

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

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Experimental program
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Embodiment 1

[0028] The preparation method of embodiment 1 medical grade type III collagen

[0029] 1. Pretreatment of newborn calf hide

[0030] Scrape off the hair and hair follicles of the fresh newborn calf hide, remove the subcutaneous tissue and fat, and finely cut it. Take 200 g of the shredded skin and add 2000 ml of a mixed solution of alcohol: haloalkane=0.5 volume ratio, soak and degrease, and replace the above solution every 5 hours , pour off the solution in 15 hours; soak it with 2000ml of 98% ethanol aqueous solution, replace once in 5 hours, and pour off the concentration in 10 hours; wash once with 2000ml of 20% ethanol aqueous solution; then use 0.1mol / L EDTA solution, pH=10.5~11.5, soak at 4°C for 6 hours, then wash 3 times with 2000ml of ultrapure water; then soak with 2000ml of 10% NaCl aqueous solution, replace once every 4 hours, pour out the solution after 8 hours, and use ultra-pure water Wash twice with 2000ml of pure water, then soak the cowhide in 2000ml of ult...

Embodiment 2

[0038] The preparation method of embodiment 2 medical grade type III collagen

[0039] 1. Pretreatment of newborn calf hide

[0040] The fresh newborn calf hide is shaved to remove the hair and hair follicles, and the subcutaneous tissue and fat are removed, and then finely cut. Get 300g shredded skin and add 5000ml of mixed solution of alcohol: haloalkane=1.5 volume ratio, soak and degrease, change the above solution every 15 hours, pour off the solution in 45 hours; soak in 5000ml of ethanol aqueous solution with a concentration of 98%, for 15 hours Replace it once, pour out the concentration in 30 hours; wash once with 5000ml of ethanol aqueous solution with a concentration of 20%; then use 0.2mol / L EDTA solution, pH=10.5~11.5, soak for 12 hours at a temperature of 6°C, and then use ultrapure water Wash 3 times with 5000ml; then soak in 5000ml of NaCl aqueous solution with a concentration of 30%, replace it once every 6 hours, pour off the solution after 12 hours, wash twi...

Embodiment 3II

[0048] The preparation method of embodiment 3 type III collagen

[0049] 1. Pretreatment of newborn calf hide

[0050] The fresh newborn calf hide is shaved to remove the hair and hair follicles, and the subcutaneous tissue and fat are removed, and then finely cut. Get 250g shredded skin and add 3000ml of mixed solution of alcohol: haloalkane=1 volume ratio, soak and degrease, change the above solution every 10 hours, pour off the solution in 30 hours; soak in 3000ml of ethanol aqueous solution with a concentration of 98% for 10 hours Replace once, pour out 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°C for 9 hours, then use ultrapure water Wash 3 times with 3000ml; then soak with 3000ml of NaCl aqueous solution with a concentration of 20%, replace once every 5 hours, pour off the solution after 10 hours, wash twice with 3000ml of ultrapure water, then soak the cowhide in 3000ml...

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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 preparation method of medical grade 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 mainly exists in tissues such as skin, blood vessels, heart, lung, liver and kidney. Type III collagen in the form of thin fibers endows tissues with strong expandability, 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 collagen type III plays an important role in Ehlers-Danlos syndrome IV (EDS IV) and the pathological study of 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 normal collagen III protein profile. Clin Genet. 2003: 63: 224-227 and Brega KE, Se...

Claims

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Application Information

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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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