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Extraction process for bovine serum albumin by graduated heat shock method

A technology for bovine serum albumin and extraction process, which is applied in the field of albumin preparation, can solve the problems of difficulty in guaranteeing albumin purity, difficult mass production, complicated production process, etc. Effect

Inactive Publication Date: 2017-11-14
天津康源生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the above-mentioned preparation method of bovine serum albumin, many high-cost salts (such as sodium citrate, sodium octanoate, etc.) and organic chemical substances (ethanol solvent, etc.) need to be added, the purity of the prepared albumin is difficult to guarantee, and the production process More complex, difficult to mass-produce

Method used

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  • Extraction process for bovine serum albumin by graduated heat shock method

Examples

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Comparison scheme
Effect test

Embodiment 1

[0020] A kind of bovine serum albumin graded heat shock method extraction process, comprises the steps:

[0021] 1. Separation of serum: Fresh whole blood from healthy cattle, defibrinated, collected by continuous centrifugation, and red blood cells are used separately;

[0022] 2. Extraction: Add serum and the same amount of normal saline into the reaction tank, stir at 20 rpm, add 0.32% sodium caprylate while stirring to fully combine with albumin; add 0.1M dilute hydrochloric acid, adjust the pH to 5.8, add Heat at 60°C for 10 hours, filter to remove the precipitate, adjust the pH to 4.5 in the clear night, heat at 60°C, keep warm for 2 hours, filter to remove the precipitate, collect the clear liquid, filter to remove the precipitate, and collect the clear liquid;

[0023] 3. Concentration, desalination and in-situ sterilization and filtration: the clear liquid is adjusted to PH to 6.8 and then sterilized by circulation, concentrated and eluted by ultrafiltration with a 10...

Embodiment 2

[0026] A kind of bovine serum albumin graded heat shock method extraction process, comprises the steps:

[0027] 1. Separation of serum: Fresh whole blood from healthy cattle, defibrinated, collected by continuous centrifugation, and red blood cells are used separately;

[0028] 2. Extraction: Add serum and the same amount of normal saline into the reaction tank, stir at 40rpm, add 0.38% sodium caprylate while stirring to fully combine with albumin; add 0.1M dilute hydrochloric acid, adjust the pH to 6.0, add Incubate at 75°C for 10 hours, filter to remove the precipitate, adjust the pH to 5.0 at night, heat at 75°C, keep warm for 3 hours, filter to remove the precipitate, and collect the supernatant;

[0029] 3. Concentration, desalination and in-situ sterilization and filtration: the clear liquid is adjusted to PH to 7.2 and then sterilized by circulation, concentrated and eluted by ultrafiltration with a 10,000 molecular weight ultrafiltration membrane, and the final concen...

Embodiment 3

[0032] A kind of bovine serum albumin graded heat shock method extraction process, comprises the steps:

[0033] 1. Separation of serum: Fresh whole blood from healthy cattle, defibrinated, collected by continuous centrifugation, and red blood cells are used separately;

[0034] 2. Extraction: Add serum and the same amount of normal saline to the reaction tank, stir at 30rpm, add 0.35% sodium caprylate while stirring to fully combine with albumin; add 0.1M dilute hydrochloric acid, adjust the pH to 6.0, add Incubate at 65°C for 10 hours, filter to remove the precipitate, adjust the pH to 5.0 in the clear night, and heat to 75°C when the pH is adjusted to 5.0, filter to remove the precipitate after 3 hours of heat preservation, and collect the supernatant;

[0035] 3. Concentration, desalination and in-situ sterilization and filtration: the clear liquid is adjusted to PH to 7.0 and then sterilized by circulation, concentrated by ultrafiltration with a 10,000 molecular weight ul...

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Abstract

The invention relates to an extraction process for bovine serum albumin by a graduated heat shock method. The process comprises the following steps: 1. serum separation: removing fibrin from healthy bovine fresh whole blood, carrying out continuous centrifugation to collect serum and erythrocytes for other use; 2. extraction: adding the serum and an equivalent volume of normal saline into a reaction tank, carrying out uniform mixing at the stirring rate of 20rpm to 40rpm, adding 0.32% to 0.38% of sodium caprylate with stirring, so as to enable the sodium caprylate to be thoroughly bonded to albumin; adding 0.1M dilute hydrochloric acid, adjusting the pH to 5.8 to 6.0, heating to the temperature of 60 DEG C to 75 DEG C, carrying out heat preservation for 10 hours, carrying out filtering to remove precipitates, adjusting the pH of supernatant to 4.5 to 5.0, carrying out heat preservation, carrying out filtering to remove precipitates, and collecting clear liquid; 3. concentrating, demineralizing and on-site sterilization filtration: adjusting the pH of the clear liquid to 6.8 to 7.2, carrying out cyclic sterilization, carrying out ultrafiltration concentrating by an ultrafiltration membrane with the molecular weight of 10,000, and carrying out saline matter eluting, wherein the final concentration of a protein concentrate does not exceed 15%; 4. carrying out freeze drying to obtain crystalline dry powder. According to the process, the used materials are simple, the cost is reduced, the process is simple and convenient, and the operation is easy.

Description

technical field [0001] The invention relates to a preparation method of albumin, in particular to a method for extracting bovine serum albumin by a graded heat shock method. Background technique [0002] Bovine serum albumin (BSA), also known as the fifth component, is a globulin in bovine serum, containing 583 amino acid residues, with a molecular weight of 66.430kDa and an isoelectric point of 4.7. , biochemical reagents widely used in various fields of pharmacy, and can also be used as medical and health food and condiment. [0003] The most commonly used preparation method of bovine serum albumin is to separate the serum from bovine blood, use ammonium sulfate for fractional precipitation, and obtain it through caprylic acid treatment; The process comprises the following steps: adopting 3.8% sodium citrate to anticoagulate blood to obtain blood plasma, heating and adding sodium caprylate, adding ethanol after adjusting the pH value, and then obtaining bovine serum album...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/765C07K1/36C07K1/34C07K1/30
CPCC07K14/765
Inventor 杨明张旭群
Owner 天津康源生物技术有限公司
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