Low-immunogenicity fish skin collagen and preparation method thereof

A technology of fish skin collagen and immunogenicity, applied in the field of collagen extraction, can solve the problems of limiting the application of collagen-based biomaterials, adverse immune reactions, etc., and achieve the effects of improving adverse reactions, simple preparation methods, and improving fever

Active Publication Date: 2021-09-28
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, substances that can cause immunogenicity in collagen, such as miscellaneous proteins, fats, telopeptides, endotoxins, etc., will cause adverse immune reactions in the human body, limiting the further application of collagen-based biomaterials

Method used

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  • Low-immunogenicity fish skin collagen and preparation method thereof
  • Low-immunogenicity fish skin collagen and preparation method thereof
  • Low-immunogenicity fish skin collagen and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] In this embodiment, the influence of the concentration of hydrogen peroxide and the number of liquid changes on the fat removal rate is determined through experiments, and the specific steps are as follows:

[0038] (1) Pre-treatment of fish skin: After the pangasius skin is thawed, remove residual meat and connective tissue, cut into 1cm pieces, wash with pre-cooled pure water, and drain;

[0039] (2) Removal of impurity protein, fat and part of endotoxin in fish skin: add 0.1M sodium hydroxide solution containing 0%, 1% and 3% hydrogen peroxide to the drained pangasius fish skin, the ratio of solid to liquid is 1:25 (w / v), after stirring for a certain period of time, change the liquid. The treatment method is to wash the hydrogen peroxide with 0.9% sodium chloride solution first, and then stir it with 0.1M sodium hydroxide solution. Conditionally change the liquid, and treat for 9 hours in total. After the treatment, wash the pangasius skin with pure water to neutrali...

Embodiment 2

[0046]In this embodiment, the influence of the solid-liquid ratio on the telopeptide removal rate in the process of removing the telopeptide is determined through experiments, and the specific steps are as follows:

[0047] (1) Pre-treatment of fish skin: After the pangasius skin is thawed, remove residual meat and connective tissue, cut into 1cm pieces, wash with pre-cooled pure water, and drain;

[0048] (2) Removal of miscellaneous protein, fat and part of endotoxin in fish skin: add 0.1M sodium hydroxide solution containing 1% hydrogen peroxide to the drained pangasius fish skin, the ratio of solid to liquid is 1:25 (w / v), stir for 3 hours; wash hydrogen peroxide with 0.9% sodium chloride solution, stir with 0.1M sodium hydroxide solution for 6 hours, change the liquid once during the period, and wash the pangasius skin with pure water until neutral;

[0049] (3) Removal of crude telopeptides from fish skin collagen: Add 0.4M acetic acid solution and 600U / g pepsin to the ...

Embodiment 3

[0055] In this embodiment, the influence of the concentration of acetic acid on the removal rate of telopeptide in the process of removing telopeptide is determined by experiment, and the specific steps are as follows:

[0056] (1) Pre-treatment of fish skin: After the pangasius skin is thawed, remove residual meat and connective tissue, cut into 1cm pieces, wash with pre-cooled pure water, and drain;

[0057] (2) Removal of impurity protein, fat and part of endotoxin in fish skin: add 0.1M sodium hydroxide solution containing 3% hydrogen peroxide to the drained pangasius fish skin, the ratio of solid to liquid is 1:25 (w / v), stir for 4.5 hours; wash hydrogen peroxide with 0.9% sodium chloride solution, add 0.1M sodium hydroxide solution and stir for 4.5 hours, after the treatment, wash pangasius skin with pure water until neutral;

[0058] (3) Crude and telopeptide removal of fish skin collagen: Add different concentrations of acetic acid solutions and 600U / g pepsin to the p...

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Abstract

The invention provides low-immunogenicity fish skin collagen and a preparation method thereof. The preparation method comprises the following steps of by taking fish skin as a raw material, pretreating the fish skin, removing impurities and degreasing, coarsely extracting collagen, removing terminal peptide, refining and drying the collagen, inactivating endotoxin and the like, thereby obtaining the collagen with obviously reduced contents of impure protein, fat, terminal peptide and endotoxin. In the fish skin pretreatment stage, hydrogen peroxide and sodium hydroxide are used for treatment, and the liquid is changed at a certain time point, so that the fish skin pretreatment effect is better; in the stages of extraction and further immunogen removal, acetic acid and protease are used for enzyme digestion terminal peptide treatment, and after a collagen finished product is obtained through refining and freeze-drying, endotoxin is inactivated through irradiation. The extraction method is simple, the extraction efficiency is relatively high, the obtained collagen immunogen substance is low in content and relatively high in purity, the adverse reaction of the collagen in the medical process is effectively reduced, and the application field of the collagen is further expanded.

Description

technical field [0001] The invention belongs to the technical field of collagen extraction, and in particular relates to a fish skin collagen with low immunogenicity and a preparation method thereof. Background technique [0002] Collagen is an extremely abundant structural protein in animals and a natural functional biomacromolecule. Procollagen molecule is the basic structural unit of collagen, which is composed of three left-handed helical α-peptide chains, and the three α-peptide chains are wound into a right-handed superhelical structure of collagen through interaction. Collagen has a tripeptide repeat sequence of (Gly-X-Y)n, where X and Y are usually proline (Pro) and hydroxyproline (Hyp). Glycine, proline and hydroxyproline work together to play an important role in forming hydrogen bonds and maintaining the triple helix structure of collagen. [0003] Collagen is mainly extracted from terrestrial animals such as cowhide, bovine Achilles tendon and pigskin. However,...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K14/78C07K1/14C07K1/30C07K1/34C07K1/36
CPCC12P21/06C07K14/78Y02A40/81
Inventor 刘楚怡侯虎马盼盼李八方王长伟杜芬
Owner OCEAN UNIV OF CHINA
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