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Preparation method of dog albumin and dog albumin product

An albumin and protein technology, which is applied to the preparation of canine albumin and the preparation of albumin, can solve the problems of unsatisfactory production practice, low albumin purity and yield, etc., and achieves low comprehensive production cost, improved purity and recovery. Efficient, easy-to-use effects

Inactive Publication Date: 2018-09-18
北京博莱得利生物技术有限责任公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Whether it is the salting out method, the organic solvent precipitation method or the heat shock method, all these methods have the defects of low albumin purity and yield, which cannot meet the needs of production practice and need to be improved urgently.

Method used

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  • Preparation method of dog albumin and dog albumin product

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The preparation of embodiment 1 canine albumin preparation

[0037] 1. Preparation method

[0038] (1) Thaw healthy canine plasma or serum, dilute with 0.03mol / L acetate buffer solution with 10 times the amount of pH 5.1, adjust the pH to 5.1 with 1.0mol / L acetic acid, and filter to clarify;

[0039] (2) Equilibrate the anion chromatography column (UniGel-80Q, purchased from Suzhou Nawei Technology Co., Ltd.) with 0.03mol / L acetate buffer solution with a pH value of 5.1 until the pH value of the effluent is 5.1, then dilute and filter Serum or plasma samples after loading;

[0040] (3) After loading the sample, rinse with 0.03mol / L acetate buffer solution with pH value of 5.1 until the absorption of A280 returns to the baseline level, and then wash with 0.03mol / L acetic acid containing 0.4mol / L NaCl Salt buffer solution (pH 6.8) elutes the chromatographic column, and this eluent (I) is the crude product of canine albumin;

[0041] (4) Equilibrate the anion chromatograp...

Embodiment 2

[0049] The preparation of embodiment 2 canine albumin preparations

[0050] 1. Preparation method

[0051] (1) Thaw healthy canine plasma or serum, dilute it with 0.03mol / L acetate buffer solution with 8 times the amount of pH 5.2, adjust the pH value to 5.2 with 1.0mol / L acetic acid, and filter to clarify;

[0052] (2) Equilibrate the anion chromatography column (UniGel-80Q, Suzhou Nawei Technology Co., Ltd.) with a 0.03mol / L acetate buffer solution with a pH value of 5.2 to a pH value of 5.2, and dilute and filter the Serum or plasma sample loading;

[0053] (3) After loading the sample, wash with 0.03mol / L acetate buffer solution with pH value of 5.2 until the absorption of A280 returns to the baseline level, and then wash with 0.03mol / L acetic acid containing 0.4mol / L NaCl Salt buffer solution (pH 7.0) elutes the chromatographic column, and this eluent (I) is the crude product of canine albumin;

[0054] (4) Equilibrate the anion chromatography column (QBestaroseFF, Ber...

Embodiment 3

[0062] The preparation of embodiment 3 canine albumin preparations

[0063] 1. Preparation method

[0064] (1) Thaw healthy canine plasma or serum, dilute tenfold with 0.03mol / L acetate buffer solution with 8 times the pH value of 4.9, adjust the pH value to 4.9 with 1.0mol / L acetic acid, and filter to clarify;

[0065] (2) Equilibrate the anion chromatography column (UniGel-80Q, Suzhou Nawei Technology Co., Ltd.) with 0.03mol / L acetate buffer solution with a pH value of 4.9 until the pH value of the effluent is 4.9, dilute and filter the Serum or plasma sample loading;

[0066] (3) After loading the sample, rinse with 0.03mol / L acetate buffer solution with a pH value of 4.9 until the absorption of A280 returns to the baseline level, and then wash with 0.03mol / L acetic acid containing 0.4mol / L NaCl Salt buffer solution (pH 7.2) elutes the chromatographic column, and this eluent (I) is the crude product of canine albumin;

[0067] (4) Equilibrate the anion chromatography col...

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Abstract

The invention discloses a preparation method of dog albumin. The preparation method comprises the following steps: (I) with serum or plasma of a healthy dog as a raw material, carrying out primary chromatographic separation by virtue of an anion exchange chromatography column UniGel-80Q, so as to obtain a dog albumin crude product; and (II) carrying out secondary chromatographic separation on thedog albumin crude product by virtue of an anion exchange chromatography column QBestaroseFF. According to the preparation method, the serum or plasma of the healthy dog is directly taken as the raw material, the chromatographic separation is carried out twice by virtue of the anion exchange chromatography columns so as to prepare the dog albumin, the content of the dog albumin in a final product is more than 100mg / ml, the purity is more than or equal to 97%, and the content of endotoxin is less than 8EU / ml. According to the preparation method, the loss of the cavity of albumin is furthest reduced, the purity and the recycling rate of the dog albumin are increased, the step is simple and convenient, the safety is high, and the comprehensive production cost is lower than that of a common low-temperature ethanol method.

Description

technical field [0001] The invention relates to a method for preparing albumin, and further relates to a method for preparing canine albumin and a canine albumin product prepared by the method, belonging to the field of canine albumin preparation. Background technique [0002] Plasma albumin is a protein with the most content in plasma, the smallest molecule, high solubility and many functions. The maintenance of plasma colloid osmotic pressure mainly depends on albumin in plasma (plasma albumin constitutes about 75% to 80% of plasma colloid osmotic pressure). The osmotic pressure produced by 1 g of albumin is equivalent to the hemodynamic effect obtained by 20 ml of plasma or 40 ml of whole blood. [0003] There are many methods for preparing albumin, mainly using salting out method, organic solvent precipitation method and heat shock method extraction. It can be obtained by salting-out method after fractional precipitation with ammonium sulfate and treatment with octanoi...

Claims

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Application Information

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IPC IPC(8): C07K14/76C07K1/18
CPCC07K14/76
Inventor 王宏伟
Owner 北京博莱得利生物技术有限责任公司
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