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Method for extracting type I collagen

A collagen, type I technology, applied in the field of medical biomaterials, can solve the problems of affecting the quality of collagen, low extraction yield, and many collagen impurities, so as to improve the extraction yield, simple preparation process, and promote collagenase The generated effect of

Inactive Publication Date: 2017-05-24
WUHAN YIJIABAO BIOMATERIAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Type I collagen currently on the market is mainly extracted from animal skin and Achilles tendon tissue. Since the type I collagen prepared by the existing extraction method is affected by the process conditions, the extraction yield is low, and the extraction There are many miscellaneous proteins contained in collagen, which seriously affects the quality of collagen, and the demand for widely used type I collagen in the fields of biomedicine and cosmetics industry is increasing day by day. Therefore, a kind of type I collagen extraction is designed It is very necessary to improve the extraction yield of collagen to meet the application of type I collagen in the fields of biomedicine and cosmetics.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment example 1

[0034] (1) Cleaning process: Clean the fresh beef Achilles tendon with ultra-pure water, remove fascia, broken bones, lean meat and fat with a scalpel blade, freeze the beef Achilles tendon in a -20°C refrigerator until hardened, and take it out Cut the bovine Achilles tendon into 1mm-thick Achilles tendon slices along the tissue fiber direction.

[0035] (2) Sterilization and degreasing process: Weigh 100g Achilles tendon slices and soak them in a mixed solution of 75% ethanol: n-hexane = 3:1 for 12 hours, the temperature of the solution is 15~25°C, stir mechanically, filter the solution with stainless steel mesh, take out Then wash 5 times with ultrapure water.

[0036] (3) Impurity protein removal process: Soak in 0.05mol / L sodium carbonate solution for 24 hours, the solution temperature is 15~25°C, mechanically stir, replace the sodium carbonate solution every 8 hours, filter the solution with stainless steel mesh, take it out and use ultra-pure Wash with water several ti...

Embodiment example 2

[0043] (1) Cleaning process: Clean the fresh beef Achilles tendon with ultra-pure water, remove fascia, broken bones, lean meat and fat with a scalpel blade, freeze the beef Achilles tendon in a -20°C refrigerator until hardened, and take it out Cut the bovine Achilles tendon into 1mm-thick Achilles tendon slices along the tissue fiber direction.

[0044](2) Sterilization and degreasing process: Weigh 100g Achilles tendon slices and soak them in a mixed solution of 75% ethanol: n-hexane = 3:1 for 12 hours, the temperature of the solution is 15~25°C, stir mechanically, filter the solution with stainless steel mesh, take out Then wash 5 times with ultrapure water.

[0045] (3) Impurity protein removal process: Soak in 0.5mol / L sodium carbonate solution for 24 hours, the solution temperature is 15~25°C, mechanically stir, replace the sodium carbonate solution every 8 hours, filter the solution with stainless steel mesh, take it out and use ultra-pure Wash with water several time...

Embodiment example 3

[0051] (1) Cleaning process: Clean the fresh beef Achilles tendon with ultra-pure water, remove fascia, broken bones, lean meat and fat with a scalpel blade, freeze the beef Achilles tendon in a -20°C refrigerator until hardened, and take it out Cut the bovine Achilles tendon into 1mm-thick Achilles tendon slices along the tissue fiber direction.

[0052] (2) Sterilization and degreasing process: Weigh 100g Achilles tendon slices and soak them in a mixed solution of 75% ethanol: n-hexane = 3:1 for 12 hours, the temperature of the solution is 15~25°C, stir mechanically, filter the solution with stainless steel mesh, take out Then wash 5 times with ultrapure water.

[0053] (3) Impurity protein removal process: Soak in 0.05mol / L sodium carbonate solution for 24 hours, the solution temperature is 15~25°C, mechanically stir, replace the sodium carbonate solution every 8 hours, filter the solution with stainless steel mesh, take it out and use ultra-pure Wash with water several ti...

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Abstract

The invention relates to a method for extracting type I collagen. The method comprises the following steps: cleaning bovine tendon, refrigerating and slicing; disinfecting and degreasing the sliced bovine tendon with 75-percent alcohol and normal hexane; adding a proper amount of active enzyme into homogenate for performing enzymatic digestion after removing impure proteins and smashing the homogenate; performing salting-out and dialysis treatment on supernatant; performing freeze drying and vacuum packaging on a collagen stock solution obtained after the dialysis to obtain a type I collagen product. The method is simple in preparation process, can be used for realizing large-scale production, and is an economical and effective extraction method. The type I collagen extracted by the method has superior biocompatibility and bioactivity, can be applied to biomedicine, and has an important application value in the field of makeups; meanwhile, epithelial cells can be activated, the hyperplasia of the epithelial cells and the generation of collagenase are facilitated, and the skin becomes tight and elastic.

Description

technical field [0001] The invention relates to the field of medical biomaterials, in particular to a method for extracting type I collagen. Background technique [0002] Collagen is the most abundant protein in mammals, accounting for 25% to 30% of the total protein in the body. Collagen is a ubiquitous component of connective tissue and an essential structural skeleton of organs and tissues. More than 27 different types of collagen have been discovered so far. Type I collagen widely exists in animal bones, corneas, skin, tendons and other tissues, with a molecular weight of about 300KD, a molecular length of about 300nm, a diameter of about 1.5nm, and a rod shape. Microstructurally, type I collagen consists of three peptide chains, including two α(I) chains and one α(II) chain. Because type I collagen has many excellent biological properties such as degradability, biocompatibility and hemostasis, it is widely used as a natural biomaterial in the fields of biomedicine and...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K14/78C07K1/36C07K1/34C07K1/30
CPCC07K14/78C12P21/06
Inventor 柯林松陈鹏唐小雄
Owner WUHAN YIJIABAO BIOMATERIAL CO LTD
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