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Biomedical mouse tail collagen extraction method

A rat tail collagen, biomedical technology, applied in peptide preparation methods, animal/human proteins, chemical instruments and methods, etc., can solve the problems of no extraction and purification process of biomedical rat tail collagen, and improve application safety , improve biocompatibility, and ensure the effect of biological functionality

Active Publication Date: 2015-07-22
HEILONGJIANG LIHAIXIN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In response to these problems, there has been no report on the extraction and purification process of biomedical rat tail collagen

Method used

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  • Biomedical mouse tail collagen extraction method
  • Biomedical mouse tail collagen extraction method
  • Biomedical mouse tail collagen extraction method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Extraction of Rat Tail Collagen for Biomedical Use

[0034] A method for extracting biomedical rat tail collagen, the steps are as follows:

[0035] (1) Take 1 kg of fresh rat tail, soak it in 1:1000 bromogeramine solution for 10-15 minutes, wash it with normal saline repeatedly for more than 5 times after taking it out; separate the rat tail tendon, and remove the residual tissue fibers of the rat tail tendon. NH 4 Cl - Soak in physiological saline solution for 24 hours, keep stirring at a low speed for 24 hours at 4°C, and change the solution twice in the middle to remove residual fat and polysaccharides. Rinse five times with deionized water, pre-freeze at -20°C for 4 hours, and freeze-dry in a vacuum for later use; the rat tails are from adult nutria, rat tails of various strains or mouse tails of various strains.

[0036] (2) Freeze-dried rat tail tendon was freeze-dried at low temperature and crushed to 200-400 mesh;

[0037] (3) Swell the rat tail tendon in 0...

Embodiment 2

[0044] A method for extracting biomedical rat tail collagen, the steps are as follows:

[0045] (1) Take 1 kg of fresh rat tail, soak it in 1:1000 bromogeramine solution for 10-15 minutes, rinse it with normal saline repeatedly for more than 5 times after taking it out; separate the rat tail tendon, and remove the residual tissue fibers of the rat tail tendon. NH 4 Cl - Soak in physiological saline solution for 24 hours, keep stirring at a low speed for 24 hours at 4°C, and change the solution twice in the middle to remove residual fat and polysaccharides. Rinse with deionized water 5 times, pre-freeze at -40°C for 2 hours, and freeze-dry in vacuum for later use;

[0046] (2), freeze-dried rat tail is crushed to 200 mesh through low-temperature freeze-drying;

[0047](3) Swell the crushed rat tail tendon in acetic acid solution in an ice-water bath, the solid-to-liquid ratio of rat tail tendon to acetic acid is 1 g: 100 mL, the concentration of acetic acid solution is 0.04 ...

Embodiment 3

[0054] A method for extracting biomedical rat tail collagen, the steps are as follows:

[0055] (1) Take 1 kg of fresh rat tail, soak it in 1:1000 bromogeramine solution for 10-15 minutes, wash it with normal saline repeatedly for more than 5 times after taking it out; separate the rat tail tendon, and remove the residual tissue fibers of the rat tail tendon. NH 4 Cl - Soak in physiological saline solution for 24 hours, keep stirring at a low speed for 24 hours at 4°C, and change the solution twice in the middle to remove residual fat and polysaccharides. Rinse with deionized water 5 times, pre-freeze at -40°C for 2 hours, and vacuum freeze-dry for later use;

[0056] (2), freeze-dried rat tail is crushed to 400 mesh through low-temperature freeze-drying;

[0057] (3) Swell the crushed rat tail tendon in acetic acid solution in an ice-water bath, the solid-liquid ratio of rat tail tendon to acetic acid is 1 g: 120 mL, the concentration of acetic acid solution is 0.1 mol / L, ...

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Abstract

The invention discloses a biomedical mouse tail collagen extraction method. A process combining an acid method with an enzymic method is adopted, a constant low-temperature aseptic environment is kept, a salt solution at the same concentration is used for twice salting out and centrifugation, so that purification steps are simplified. Adopted acid is a low-concentration acid solution, and citric acid is used for purification instead of acetic acid. From raw material extraction to sample generation, vacuum freeze drying is performed for many times, collagen micro powder with water content of lower than 3 percent is obtained by further treatment, and is finally sterilized and aseptically packaged, and the purity of a collagen powder finished product is over 98 percent. According to the method, the immunogenicity of the collagen is reduced, yield and biocompatibility are improved, the cost is reduced, the method is applied to a biomedical material, and a peculiar triple helix structure of the obtained collagen can be completely preserved.

Description

technical field [0001] The invention relates to a method for extracting biomedical rat tail collagen. Background technique [0002] Collagen is the main component of the extracellular matrix, and it is the most abundant and widely distributed protein in animals. Collagen has a unique triple helix structure and contains a unique hydroxyproline, which has good biocompatibility and biodegradability. It is closely related to the formation and maturation of tissues, the transmission of information between cells, joint lubrication, wound healing, calcification, blood coagulation and aging. Rat tail collagen is type I collagen, and type I collagen is widely found in tendon, skin, bone and other tissues of animals. Type I collagen has the characteristics of self-assembly into fibers, which can be used in cell culture and three-dimensional scaffold materials in tissue engineering, so that cells can grow in a close to real environment. At present, the extraction methods of collagen...

Claims

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Application Information

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IPC IPC(8): C07K14/78C07K1/18C07K1/14
CPCC07K14/78
Inventor 张志平杨宁
Owner HEILONGJIANG LIHAIXIN BIOTECH CO LTD
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