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Method for separating and purifying molecule recombination collagen protein

A recombinant collagen, separation and purification technology is applied in the field of separation and purification of collagen for molecular recombination, and achieves the effects of high solubility, high biological safety and low price

Inactive Publication Date: 2008-04-09
SHANGHAI FISHERIES UNIV
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  • Abstract
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Problems solved by technology

[0003] Collagen has significant functional properties as a biomedical material, but there are also deficiencies. While maintaining the biological activity of collagen through physical cross-linking, chemical cross-linking and blending with other polymer materials, new functions are given. In recent years, composite materials formed by co-hybridization of different natural polymers have shown advantages and properties that other materials cannot replace, and are currently a hot spot in the research and development of biomaterials.

Method used

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Examples

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Embodiment Construction

[0015] Under mild extraction conditions, the method of separating and purifying collagen for molecular reorganization from blue shark scalp is as follows:

[0016] (1) Soak 30g of frozen blue shark scalp in phosphate buffer (10mM, pH 7.4-7.6) to remove scales and residual meat, cut into small pieces of 0.5cm×0.5cm, then soak and rinse with deionized water 3 to 5 times.

[0017] (2) The cleaned fish skin was soaked in 250ml of phosphate buffer (100mM, pH7.4-7.6) for 72 hours, then homogenized for 5min, centrifuged at 4°C and 10000rpm for 15min, and the supernatant was taken.

[0018] (3) Precipitation Repeat the process of (2) once, combine the supernatants, add ammonium sulfate to a saturation of 70%-80%, centrifuge (8000rpm, 4°C, 15min), and retain the precipitate.

[0019] (4) Dissolve the precipitate in about 20 ml of phosphate buffer (200 mM, pH 7.0, 1.5 g / ml).

[0020] (5) Using phosphate buffer (200mM, pH 7.0) as the eluent, chromatograph the above solution with dextra...

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Abstract

The present invention pertains to the technical field of collagens, in particular to a method for separation, purification and recombination of the collagens. The present invention takes the shark scalps as the raw materials, the soaking, the salt precipitation and the chromatography in the phosphate buffer solutions with the different concentrations are carried out sequentially for extraction and purification of the collagens. The obtained collagens can better maintain the spatial structural features of the collagens which exists naturally, the collagens can be prepared into the biomedical engineering materials with the good anti-leakage capacity and long degradation time after the molecular recombination, which can be used as various medical surgical materials with the higher requirements for reconstruction of veins and arteries and so on. The method of the present invention has rich sources, low price, simple process and no environmental pollution.

Description

technical field [0001] The invention belongs to the technical field of collagen, and in particular relates to a method for separating and purifying collagen for molecular recombination. Background technique [0002] Collagen is a structural protein that exists in the intercellular matrix and is widely distributed in animal connective tissues. Collagen fibers composed of collagen make animal connective tissue have good elasticity, ductility and toughness. Collagen has basic unit amino acids that are easy to design and modify, has no cytotoxicity, can regulate biodegradation speed, can specifically promote or inhibit cell-material interaction, can cause minimal immune response and inflammation, and is easy to purify and process. It has many advantages such as excellent biochemical compatibility with aqueous solution and physiological conditions. Natural biomaterial collagen has broad application prospects in the field of tissue engineering. Collagen obtained from the connect...

Claims

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Application Information

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IPC IPC(8): C07K1/36C07K1/12C07K1/16C07K14/47
Inventor 吴文惠朱夕波张艳包斌王南平李燕缪辉南
Owner SHANGHAI FISHERIES UNIV
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