A high-purity collagen extraction process

A technology of collagen and extraction process, which is applied in the field of protein extraction, can solve the problems of no virus process verification, collagen cannot be applied to tissue engineering, and no research on the integrity of collagen triple helix structure, so as to achieve the effect of improving purity

Active Publication Date: 2022-03-11
浙江崇山生物制品有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Chinese patent application 201811465406.1 discloses a method for extracting collagen, including: ① thaw the bovine Achilles tendon, remove blood, surface foreign matter and fascia, and perform a pretreatment with the solution; Use the solution for secondary pretreatment; ③The bovine Achilles tendon treated in step 2 is frozen and sliced, and pretreated with the solution; ④The pretreated bovine Achilles tendon slice is continuously enzymatically hydrolyzed, and the supernatant is obtained by centrifugation to prepare a crude collagen extract ; ⑤ Adjust the pH of the crude collagen extract, add neutral salt for salting out, and centrifuge to obtain the collagen precipitate; ⑥ Add phosphate buffer to dissolve the above precipitate to obtain a specific concentration of collagen. In this application, the pretreatment step was carried out twice. The purpose is to remove all the fat tissue, etc., and the reagents used in the pretreatment are ethanol solution, sodium hypochlorite, EDTA-Na 2 , TritionX-114, tributyl phosphate, Triton X-100, sodium hydroxide solution, sodium carbonate solution, the application only focused on the purity of collagen, but did not study the integrity of the triple helix structure of collagen, and Without virus process verification, the extracted collagen cannot be applied in the field of tissue engineering

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  • A high-purity collagen extraction process
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  • A high-purity collagen extraction process

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1 A kind of high-purity collagen protein extraction process

[0054] Include the following steps:

[0055] (1) Achilles tendon pretreatment step;

[0056] Described Achilles tendon pretreatment step also includes following sub-steps:

[0057] ①Thaw the Achilles tendon, remove the blood and superficial fascia, wash it with purified water, soak it in 70% alcohol for 10 minutes, then rinse it with purified water three times, put it in the refrigerator, and freeze it for later use;

[0058] ②Take out the Achilles tendon after it is completely frozen, remove the residual fat film on the surface of the Achilles tendon, and then slice it to obtain slices of the Achilles tendon;

[0059] ③Achilles tendon slices were taken and processed as follows:

[0060] Step 1: Soak with 0.2% peracetic acid for 5 minutes, then wash with purified water three times;

[0061] Step 2: Repeat step a to process once;

[0062] Step 3: Soak in a mixed solution of 0.2% tributyl phosph...

Embodiment 2

[0070] Embodiment 2 A kind of high-purity collagen extraction process

[0071] Include the following steps:

[0072] (2) Achilles tendon pretreatment step;

[0073] Described Achilles tendon pretreatment step also includes following sub-steps:

[0074] ①Thaw the Achilles tendon, remove the blood and superficial fascia, wash it with purified water, soak it in 75% alcohol for 3 minutes, then rinse it with purified water three times, put it in the refrigerator, and freeze it for later use;

[0075] ②Take out the Achilles tendon after it is completely frozen, remove the residual fat film on the surface of the Achilles tendon, and then slice it to obtain slices of the Achilles tendon;

[0076] ③Achilles tendon slices were taken and processed as follows:

[0077] Step 1: Soak in 0.5% peracetic acid for 2 minutes, then wash with purified water three times;

[0078] Step 2: Repeat step a to process once;

[0079] Step 3: Soak in a mixed solution of 0.5% tributyl phosphate and 0.5...

Embodiment 3

[0087] Embodiment 3 A kind of high-purity collagen extraction process

[0088] Include the following steps:

[0089] (1) Achilles tendon pretreatment step;

[0090] Described Achilles tendon pretreatment step also includes following sub-steps:

[0091] ①Thaw the Achilles tendon, remove the blood and superficial fascia, wash it with purified water, soak it in 75% alcohol for 8 minutes, then rinse it with purified water three times, put it in the refrigerator, and freeze it for later use;

[0092] ②Take out the Achilles tendon after it is completely frozen, remove the residual fat film on the surface of the Achilles tendon, and then slice it to obtain slices of the Achilles tendon;

[0093] ③Achilles tendon slices were taken and processed as follows:

[0094] Step 1: Soak in 0.4% peracetic acid for 3 minutes, then wash with purified water three times;

[0095] Step 2: Repeat step a to process once;

[0096] The third step: Soak with 0.4% tributyl phosphate and 0.2% Tween 80...

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Abstract

The invention provides a high-purity collagen extraction process, which belongs to the technical field of protein extraction. The extraction process includes the Achilles tendon pretreatment step and the collagen extraction and purification step. During the implementation process, the reagents in the pretreatment process, as well as the concentration of various reagents, mixing methods and addition sequences are reasonably controlled, and the pretreatment process is obviously improved. The removal efficiency of Achilles tendon fat during the process improves the purity and activity of collagen, and the obtained collagen has a complete triple helix structure, which is safer and can meet the needs of cosmetics, medical devices, and tissue engineering.

Description

technical field [0001] The invention belongs to the technical field of protein extraction, and in particular relates to a process for extracting high-purity collagen from Achilles tendon. Background technique [0002] Collagen is the most abundant type of protein in animals, accounting for more than 30% of the total protein content, and is widely distributed in tissues and organs such as bones, cartilage, tendons, skin, and blood vessels. Among the 28 known collagen types, type I collagen is the most abundant and widely distributed. Because of its good biocompatibility, low immunogenicity, good biomechanical properties, etc., it can be used as a natural biological framework component, and has huge application space in clinical medicine, tissue engineering, medical bioengineering and other fields. [0003] There are many existing collagen extraction methods, but almost all of them have a problem of removing impurities. Removing lipids by adding organic solvents such as aceto...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/78C07K1/36C07K1/34C07K1/30C07K1/14
CPCC07K14/78
Inventor 夏伟张智武唐冬慧
Owner 浙江崇山生物制品有限公司
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