CN107344965A - A kind of bovine serum albumin(BSA) indexes heat shock extraction process - Google Patents
A kind of bovine serum albumin(BSA) indexes heat shock extraction process Download PDFInfo
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- CN107344965A CN107344965A CN201610290629.3A CN201610290629A CN107344965A CN 107344965 A CN107344965 A CN 107344965A CN 201610290629 A CN201610290629 A CN 201610290629A CN 107344965 A CN107344965 A CN 107344965A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/76—Albumins
- C07K14/765—Serum albumin, e.g. HSA
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Abstract
A kind of bovine serum albumin(BSA) indexes heat shock extraction process, comprises the following steps：1. separate serum：Healthy ox fresh whole blood, is defibrinated, and continuous centrifugal collects serum, and red blood cell is separately used；2. extraction：Serum and the physiological saline of equivalent are added into retort, 20 40rpm are stirred and evenly mixed, and are added 0.32 0.38% Sodium Caprylate while stirring, are allowed to fully be combined with albumin；0.1M watery hydrochloric acid is added, adjusts PH to 5.8 6.0,60 DEG C 75 DEG C of heating is incubated 10 hours, filters off precipitation, and the stillness of night adjusts PH to 4.5 5.0, insulation, filters off precipitation, collects clear liquid；3. concentration, desalination and aseptic filtration in place：Clear liquid adjusts PH to 6.8 7.2 degerming through circulating, and is concentrated by ultrafiltration with 10,000 molecular weight milipore filters, elution salinity, and protein concentrated solution final concentration is no more than 15%；4. freeze-drying is lenticular dry powder.Materials of the present invention are simple, cost-effective, simple process, easily operated.
Description
Technical field
The present invention relates to the preparation method of albumin, more particularly to a kind of bovine serum albumin(BSA) indexing heat shock extraction process.
Background technology
Bovine serum albumin(BSA) (BSA), also known as BSA, it is a kind of globulin in cow's serum, include 583 amino acid residues, molecular weight is 66.430kDa, isoelectric point is 4.7, is biology subjects and doctor, the wide variety of biochemical reagents in each field of pharmacy, it is also possible to make medicines and health protection food, flavouring.
The most frequently used preparation method of bovine serum albumin(BSA) is to isolate serum by raw material of ox blood, after ammonium sulfate precipitation, refines and obtains through octanoic acid processing；Chinese patent literature CN 1544470C disclose a kind of bovine serum albumin hot ethanol extracting process, its method is to obtain blood plasma using 3.8% sodium citrate anticoagulation, heating adds Sodium Caprylate, adjusts after pH value plus ethanol, then filtered, desalination, concentration, it is lyophilized after bovine serum albumin(BSA).Need to add the higher salt of many costs (such as sodium citrate, Sodium Caprylate) and organic chemicals (alcohol solvent etc.) in above-mentioned bovine serum albumin(BSA) preparation method, the purity for preparing albumin is difficult to ensure that, and production technology is more complicated, it is difficult to is produced in enormous quantities.
The content of the invention
It is an object of the invention in place of overcome the deficiencies in the prior art, there is provided a kind of bovine serum albumin(BSA) indexing heat shock extraction process that a kind of purity is high, technological process is simple, cost is low.
The purpose of the present invention is achieved through the following technical solutions：
A kind of bovine serum albumin(BSA) indexes heat shock extraction process, comprises the following steps：
1. separate serum：Healthy ox fresh whole blood, is defibrinated, and continuous centrifugal collects serum, and red blood cell is separately used；
2. extraction：Serum and the physiological saline of equivalent are added into retort, 20-40rpm is stirred and evenly mixed, and is added 0.32-0.38% Sodium Caprylates while stirring, is allowed to fully be combined with albumin；0.1M watery hydrochloric acid is added, adjusts PH to 5.8-6.0,60 DEG C -75 DEG C of heating is incubated 10 hours, filters off precipitation, and the stillness of night adjusts PH to 4.5-5.0, insulation, filters off precipitation, collects clear liquid；
3. concentration, desalination and aseptic filtration in place：Clear liquid adjusts PH to 6.8-7.2 degerming through circulating, and is concentrated by ultrafiltration with 10,000 molecular weight milipore filters, elution salinity, and protein concentrated solution final concentration is no more than 15%；
4. freeze-drying is lenticular dry powder.
Moreover, 60 DEG C -75 DEG C can be heated when the stillness of night adjusting PH to 4.5-5.0 in the step 2, precipitation is filtered off after being incubated 2-3 hours, collects clear liquid.
Advantages of the present invention and economic effect are：
1. in technical process, 60 DEG C -75 DEG C are incubated 10 hours, can effectively inactivation of viruses class exogenous factor, the security of product it is secure；Thorough inactivated proteases simultaneously, avoid the label of user impacted.
2. indexing heating, alleviating coprecipitated situation in albuminous degeneration separation process, albumin yield improves, and obtaining albumin per 100ml bovine serums is no less than 2g.
It is 3. in place degerming so that protein liquid avoids bacteria breed in the concentration of long period, desalination processes, the problem that excluding bacterium metabolite.
4. materials are simple, cost-effective, simple process, easily operated.
Brief description of the drawings
Fig. 1 is the process chart of the present invention.
Embodiment
For present disclosure, feature and effect can be further appreciated that, following examples are hereby enumerated, and it is as follows to coordinate accompanying drawing to describe in detail.It should be noted that the present embodiment is descriptive, it is not limited, it is impossible to thus limit protection scope of the present invention.
Embodiment 1：
A kind of bovine serum albumin(BSA) indexes heat shock extraction process, comprises the following steps：
1. separate serum：Healthy ox fresh whole blood, is defibrinated, and continuous centrifugal collects serum, and red blood cell is separately used；
2. extraction：Serum and the physiological saline of equivalent are added into retort, 20rpm is stirred and evenly mixed, added 0.32% Sodium Caprylate while stirring, be allowed to fully be combined with albumin；0.1M watery hydrochloric acid is added, adjusts PH to 5.8,60 DEG C is heated and is incubated 10 hours, filters off precipitation, the stillness of night adjusts PH to 4.5, heats 60 DEG C, and insulation filters off precipitation after 2 hours, collect clear liquid, filters off precipitation, collects clear liquid；
3. concentration, desalination and aseptic filtration in place：Clear liquid adjusts PH to 6.8 degerming through circulating, and is concentrated by ultrafiltration with 10,000 molecular weight milipore filters, elution salinity, and protein concentrated solution final concentration is no more than 15%；
4. freeze-drying is lenticular dry powder.
Embodiment 2：
A kind of bovine serum albumin(BSA) indexes heat shock extraction process, comprises the following steps：
1. separate serum：Healthy ox fresh whole blood, is defibrinated, and continuous centrifugal collects serum, and red blood cell is separately used；
2. extraction：Serum and the physiological saline of equivalent are added into retort, 40rpm is stirred and evenly mixed, added 0.38% Sodium Caprylate while stirring, be allowed to fully be combined with albumin；0.1M watery hydrochloric acid is added, adjusts PH to 6.0,75 DEG C is heated and is incubated 10 hours, filters off precipitation, the stillness of night adjusts PH to 5.0, heats 75 DEG C, and insulation filters off precipitation after 3 hours, collect clear liquid；
3. concentration, desalination and aseptic filtration in place：Clear liquid adjusts PH to 7.2 degerming through circulating, and is concentrated by ultrafiltration with 10,000 molecular weight milipore filters, elution salinity, and protein concentrated solution final concentration is no more than 15%；
4. freeze-drying is lenticular dry powder.
Embodiment 3：
A kind of bovine serum albumin(BSA) indexes heat shock extraction process, comprises the following steps：
1. separate serum：Healthy ox fresh whole blood, is defibrinated, and continuous centrifugal collects serum, and red blood cell is separately used；
2. extraction：Serum and the physiological saline of equivalent are added into retort, 30rpm is stirred and evenly mixed, added 0.35% Sodium Caprylate while stirring, be allowed to fully be combined with albumin；0.1M watery hydrochloric acid is added, adjusts PH to 6.0,65 DEG C is heated and is incubated 10 hours, filters off precipitation, the stillness of night adjusts PH to 5.0, and 75 DEG C can be heated when adjusting PH to 5.0, and insulation filters off precipitation after 3 hours, collect clear liquid；
3. concentration, desalination and aseptic filtration in place：Clear liquid adjusts PH to 7.0 degerming through circulating, and is concentrated by ultrafiltration with 10,000 molecular weight milipore filters, elution salinity, and protein concentrated solution final concentration is no more than 15%；
4. freeze-drying is lenticular dry powder.
The assay of product of the present invention：
。
Claims (2)
1. a kind of bovine serum albumin(BSA) indexes heat shock extraction process, it is characterised in that：Comprise the following steps：
(1) separates serum：Healthy ox fresh whole blood, is defibrinated, and continuous centrifugal collects serum, and red blood cell is separately used；
(2) is extracted：Serum and the physiological saline of equivalent are added into retort, 20-40rpm is stirred and evenly mixed, added while stirring
0.32-0.38% Sodium Caprylates, it is allowed to fully be combined with albumin；0.1M watery hydrochloric acid is added, adjusts PH to 5.8-6.0, heating
60 DEG C -75 DEG C are incubated 10 hours, filter off precipitation, and the stillness of night adjusts PH to 4.5-5.0, insulation, filters off precipitation, collects clear
Liquid；
(3) concentrations, desalination and aseptic filtration in place：Clear liquid adjusts PH to 6.8-7.2 degerming through circulating, with 10,000 molecular weight ultrafiltration
Film is concentrated by ultrafiltration, elution salinity, and protein concentrated solution final concentration is no more than 15%；
(4) freeze-dryings are lenticular dry powder.
A kind of 2. bovine serum albumin(BSA) indexing heat shock extraction process according to claim 1, it is characterised in that：It is described
60 DEG C -75 DEG C can be heated when the stillness of night adjusting PH to 4.5-5.0 in step 2, precipitation is filtered off after being incubated 2-3 hours, is collected
Clear liquid.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610290629.3A CN107344965A (en) | 2016-05-05 | 2016-05-05 | A kind of bovine serum albumin(BSA) indexes heat shock extraction process |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109734796A (en) * | 2019-02-01 | 2019-05-10 | 广州蕊特生物科技有限公司 | A kind of technique separating albumin from haemolysis serum |
| CN112521488A (en) * | 2020-12-26 | 2021-03-19 | 郑州伊美诺生物技术有限公司 | Preparation method of bovine serum albumin |
-
2016
- 2016-05-05 CN CN201610290629.3A patent/CN107344965A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109734796A (en) * | 2019-02-01 | 2019-05-10 | 广州蕊特生物科技有限公司 | A kind of technique separating albumin from haemolysis serum |
| CN109734796B (en) * | 2019-02-01 | 2022-04-15 | 广州蕊特生物科技有限公司 | Process for separating albumin from haemolytic serum |
| CN112521488A (en) * | 2020-12-26 | 2021-03-19 | 郑州伊美诺生物技术有限公司 | Preparation method of bovine serum albumin |
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Application publication date: 20171114 |