CN107216383A - A kind of preparation method of human serum albumin and the human serum albumin - Google Patents
A kind of preparation method of human serum albumin and the human serum albumin Download PDFInfo
- Publication number
- CN107216383A CN107216383A CN201710568707.6A CN201710568707A CN107216383A CN 107216383 A CN107216383 A CN 107216383A CN 201710568707 A CN201710568707 A CN 201710568707A CN 107216383 A CN107216383 A CN 107216383A
- Authority
- CN
- China
- Prior art keywords
- serum albumin
- human serum
- human
- finished product
- content
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Zoology (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Toxicology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The human serum albumin that the present invention provides a kind of preparation method of human serum albumin and prepared with this method.The preparation method of this kind of human serum albumin comprises the following steps：Step 1: using cold ethanol method from human plasma the supernatant of extraction components IV as raw material；Step 2: carrying out refined filtration processing to the supernatant of component IV；Step 3: refined filtration processing obtains clarified solution, regulation pH obtains component V and precipitated；Step 4: purification process obtains human serum albumin finished product after the precipitation dissolving of component V.12IU/ml should be not higher than by obtaining Prekallikrein content in human serum albumin finished product with the inventive method, and Content of polymer is not higher than 2%.With easy to operate, process stabilizing in production application, the quality standard of final products is higher than national standard.
Description
Technical field
The invention belongs to biological product technical field, it is related to a kind of preparation method of human serum albumin and is prepared with this method
Human serum albumin.
Background technology
Human serum albumin is the protein formulation extracted from human normal plasma, and human serum albumin is mainly used in because losing blood, creating
Injure and suffered a shock caused by burn etc., the cerebral caused by encephaledema and brain damage increases, to preventing and treating Hypoproteinemia and hepatic sclerosis
It is the indispensable medicine healed the wounded and rescue the dying or oedema caused by nephrosis or ascites have preferable curative effect.The country mainly uses low temperature at present
Ethanol is isolated and purified, and different manufacturers are slightly different using the parameter and processing step of Low-temperature Ethanol Processes, therefore
Human blood albumin products quality on the market is variant.Prekallikrein (PKA) and Content of polymer are people
The biggest factor of adverse reaction is produced in blood albumin preparation clinical practice, therefore, how in preparation technology, reduction is explored
PKA and Content of polymer method, are always the emphasis that practitioner pays special attention to.
Plasma coagulation factors Ⅻ, into after Ⅻ a, activates kallikreinogen (PK) through surface activation, generates kallikrein
(K), K makes kininase discharge slow kinases, causes vasodilation, the increase of capillary permeability and then drop in blood pressure etc. bad anti-
Should.The fragment of the factor Ⅻ of these activation is just called Prekallikrein (PKA).Clinically using containing a large amount of PKA
During blood product (such as human serum albumin), patients' blood's decline, flushed face, expiratory dyspnea etc. can be caused bad anti-
Should.There is corresponding mandatory provision to this national correlation department：《Chinese Pharmacopoeia》The three regulation human serum albumin finished products of version in 2015
In " Prekallikrein (PKA) " content should be not higher than 35IU/ml,
In normal human blood and in the absence of macroaggregates of albumin, because the albumin products containing polymer are injected into human body
Afterwards, albumin colloid osmotic drops in blood can be caused, excretion is accelerated in vivo, therefore many polymer content mistakes of Clinical practice
High albumin products, by the safe and effective of shadow medicine.《Chinese Pharmacopoeia》In the three regulation human serum albumin finished products of version in 2015
" Content of polymer " should be not higher than 5%.
The content of the invention
Prekallikrein in human serum albumin finished product can be effectively reduced it is an object of the invention to provide one kind
(PKA) and polymer albumin preparation method.
To achieve the above object, the present invention uses following technical scheme：
The present invention provides a kind of human serum albumin preparation method, and this method comprises the following steps：Step 1: with cold ethanol
Method supernatant of extraction components IV from human plasma is raw material；Step 2: carrying out refined filtration processing to the supernatant of component IV；Step 3:
The clarified solution that refined filtration processing is obtained is adjusted to 4.80 ± 0.1 acquisition groups with 0.5mol/L acetic acid (containing 40% ethanol) solution regulation pH
Divide V precipitation；Step 4: purification process obtains human serum albumin finished product after the precipitation dissolving of component V；
Refined filtration processing method is carried out to the supernatant of component IV wherein described in step 2,
(1) cooling is handled：The supernatant of component IV is down to-6-- 10 DEG C；
(2) kieselguhr adsorption：Diatomite is added, dosage is 2~10g/L, stirring and adsorbing time >=30 minute；
(3) filter plate refined filtration：Refined filtration is carried out with filter plate, environment temperature control is below 10 DEG C during filtering, and filter pressure does not surpass
0.3MPa is crossed, clarified solution after filtering is collected；
Wherein, purification process obtains human serum albumin finished product detailed process bag after the precipitation dissolving of component V described in step 4
Include following steps：
(1) purify：30/95 (L), which is precipitated, by final per kilogram adds 95% ethanol and 0 DEG C of water for injection to the precipitation weights of F V
5 times of volumes of amount are stirred dissolving, then adjust pH to 4.50~4.70, and control temperature was stirred at -2~-3 DEG C
Filter.
(2) ultrafiltration：Concentrated with 10KD film bag ultrafilters, then with 1% and 0.7% sodium chloride solution respectively to product
The dialysis that the dialysis of the weight such as progress is 4 times and 5 times.
(3) match somebody with somebody liquid, according to product specification prepare corresponding protein content, Na+ contents≤150mmol/L, K+ content≤
2mmol/L, sad sodium content are about that 0.160 ± 0.02mmol/g. protein, pH value are 6.8~7.0；
(4) pasteurization, pasteurization temperature is 60 ± 0.5 DEG C, and the pasteurization time is 10 hours；
(5) degerming packing, degerming packing is carried out with degerming filter core according to product specification
The present invention also provides a kind of human serum albumin obtained by the above method, wherein, in the human serum albumin finished product
Prekallikrein (PKA) content should be not higher than 12IU/ml；Content of polymer is not higher than 2%；Human serum albumin only with
Anti-human blood plasma produces precipitation line, and precipitation line is not produced with anti-horse, anti-ox, anti-pig, anti-sheep blood plasma；Compared with human normal plasma, it is main
It is albumin to want precipitation line.
The invention has the advantages that：
1st, easy to operate in production application, process stabilizing does not interfere with the recovery rate of final products.2nd, kassinin kinin is released in finished product
Put zymoexcitator (PKA) content ratio《Chinese Pharmacopoeia》Version reduction by 60% in 2015, Content of polymer ratio《Chinese Pharmacopoeia》2015
Year version reduction by 60%, can be greatly improved the security and validity of human serum albumin product, with good clinical value.
Embodiment
With reference to specific embodiment to the present invention.
Embodiment 1：
1. blood plasma is obtained
Human normal plasma is gathered by blood-collecting station of our company
2. blood plasma is got
By production ordering requirement, requirement 7983.585kg qualified blood plasma is extracted.
3. blood plasma merges and melted
In D grades of clean factories, 37 DEG C of control dissolving water temperature ＜ melts method using water-bath and carries out blood plasma thawing.Blood after thawing
4 DEG C of slurry temperature degree ＜.
4. the ethanol precipitation of components I+II+III and it is separated by filtration
It is dilute by 0.1 times of 0.9%NaCl solution of Plasma volumes after the accurate metered volume 7549L of blood plasma after thawing
Release to 8305L, pH6.3 adjusted with pH4.0 buffer solutions, less than -15 DEG C of 95% ethanol 2216L of addition, final temperature control -
5℃；Diatomite is added by 20g/L reaction solutions respectively in reaction solution, separation of solid and liquid is carried out with BECO companies filter plate, component is obtained
I+II+III supernatant.
5. the supernatant of component IV is obtained
By the supernatant 13459L of components I+II+III, pH6.15,95% second of less than -15 DEG C of addition are adjusted with pH4.0 buffer solutions
Alcohol 4895L, final temperature is controlled at -5 DEG C；Diatomite is added by 15g/L reaction solutions respectively in reaction solution, is filtered with BECO companies
Plate carries out separation of solid and liquid, obtains the supernatant of component IV.
6. the processing of the refined filtration of component IV
Component IV is cooled to -6~-10 DEG C, the diatomite of Yi Rui stones company near a river is added, dosage is 2~10g/L, is stirred
Adsorption time >=30 minute are mixed, with BECO companies filter plate to carrying out refined filtration, environment temperature control is below 10 DEG C during filtering, filtering
Pressure is no more than 0.3MPa, collects clarified solution after filtering.
7. the precipitation of component V is obtained
The clarified solution 21201L of acquisition is adjusted to 4.80 with 0.5mol/L acetic acid (contain 40% ethanol) solution regulation pH ±
0.1, -9~-10 DEG C are cooled to, with BECO companies filter plate to carrying out separation of solid and liquid, component V is obtained and precipitates.
8. human serum albumin is obtained
Component V is precipitated into 1047.8kg, precipitating 30/95 (L) by final per kilogram adds 95% ethanol and 0 DEG C of water for injection
5 times of volumes to the Sediment weights of F V are stirred dissolving, and after fully dissolving 2 hours, lysate is cooled to -3 DEG C, public with BECO
Take charge of filter plate to carry out separation of solid and liquid, supernatant after filtering is concentrated with 10KD film bag ultrafilters, then with 1% and 0.7% chlorine
Change sodium solution and the weight such as carry out to product respectively and dialyse the dialysis of 4 times and 5 times, collect ultrafiltrate and carry out matching somebody with somebody liquid, be finally formulated as：
Protein content 20%, Na+ content 142mmol/L, K+ contents 0.1mmol/L, octanoic acid sodium content 0.158mmol/g. protein, pH
Value 6.8.
It will be carried out with the protein liquid after liquid after 60 DEG C of continuous 10 hours constant temperature pasteurizations, pasteurization, aseptic filtration point
Dress, obtains human serum albumin finished product.
9. detection
According to《Chinese Pharmacopoeia》Version three in 2015, samples 32 bottles, is detected, as a result following as shown in table 1：
The finished product human serum albumin testing result of table 1
It is recognised that the illustrative embodiments that above-described embodiment uses only for explanation inventive principle, but this hair
Bright to be not limited only to this, those skilled in the art can make various improvement and change in the case where not departing from real situation of the present invention, this
A little improvement and change fall within protection scope of the present invention.
Claims (6)
1. a kind of reduce the processing method of human blood albumin products Prekallikrein, it is characterised in that：Step 1: with
Cold ethanol method supernatant of extraction components IV from human plasma is raw material；Step 2: carrying out refined filtration processing to the supernatant of component IV；
Step 3: the clarified solution that refined filtration processing is obtained is adjusted to the pH containing the regulation of the ethanol 0.5mol/L acetums of volume fraction 40%
4.80 ± 0.1 acquisition components V are precipitated；Step 4: purification process obtains human serum albumin finished product after the precipitation dissolving of component V.
2. the processing method of human blood albumin products Prekallikrein is reduced described in claim 1, it is characterised in that
Step 2 comprises the following steps：
(1) cooling is handled：The supernatant of component IV is down to-6-- 10 DEG C；
(2) kieselguhr adsorption：Diatomite is added, dosage is 2~10g/L, stirring and adsorbing time >=30 minute；
(3) filter plate refined filtration：Refined filtration is carried out with filter plate, environment temperature control is below 10 DEG C during filtering, and filter pressure is no more than
0.3MPa, collects clarified solution after filtering.
3. the processing method of human blood albumin products Prekallikrein is reduced described in claim 1, it is characterised in that
Step 4 comprises the following steps：
(1) purify：95% ethanol and 0 DEG C of water for injection are added to the 5 of components precipitate weight by final per kilogram precipitation 30/95L
Times volume is stirred dissolving, then adjusts pH to 4.50~4.70, and control temperature is stirred filtering at -2~-3 DEG C.
(2) ultrafiltration：Concentrated, then product is carried out respectively with 10KD film bag ultrafilters with 1% and 0.7% sodium chloride solution
Dialysis of 4 times and 5 times etc. weight dialysis.
(3) match somebody with somebody liquid, protein content, Na+ contents≤150mmol/L, K+ content≤2mmol/L, octanoic acid are prepared according to product specification
Sodium content be about 0.160 ± 0.02mmol/g. protein, pH value be 6.8~7.0；
(4) pasteurization, pasteurization temperature is 60 ± 0.5 DEG C, and the pasteurization time is 10 hours；
(5) degerming packing, degerming packing is carried out with degerming filter core according to product specification.
4. the human serum albumin finished product obtained by any one of the claim 1-3 processing methods, it is characterised in that：The human blood
Prekallikrein content should be not higher than 12IU/ml in albumin finished product.
5. the human serum albumin finished product described in claim 4, it is characterised in that：Content of polymer is not in the human serum albumin finished product
Higher than 2%.
6. the human serum albumin finished product described in any one of claim 5, it is characterised in that：The human serum albumin only with anti-human blood
Slurry produces precipitation line, and precipitation line is not produced with anti-horse, anti-ox, anti-pig, anti-sheep blood plasma；Compared with human normal plasma, major precipitation
Line is albumin.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710568707.6A CN107216383A (en) | 2017-07-13 | 2017-07-13 | A kind of preparation method of human serum albumin and the human serum albumin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710568707.6A CN107216383A (en) | 2017-07-13 | 2017-07-13 | A kind of preparation method of human serum albumin and the human serum albumin |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107216383A true CN107216383A (en) | 2017-09-29 |
Family
ID=59952886
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710568707.6A Pending CN107216383A (en) | 2017-07-13 | 2017-07-13 | A kind of preparation method of human serum albumin and the human serum albumin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107216383A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108864278A (en) * | 2018-07-27 | 2018-11-23 | 珠海宝锐生物科技有限公司 | A method of preparing molecular biosciences grade bovine serum albumin(BSA) |
| CN110317261A (en) * | 2018-03-28 | 2019-10-11 | 发贵科技（贵州）有限公司 | Improved two steps ultrafiltration produces human serum albumin |
| CN110878120A (en) * | 2019-10-31 | 2020-03-13 | 华兰生物工程重庆有限公司 | Method for recovering albumin from component II supernatant |
| CN116589559A (en) * | 2023-06-05 | 2023-08-15 | 广东丹霞生物制药有限公司 | Process for preparing human serum albumin |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4440679A (en) * | 1980-03-05 | 1984-04-03 | Cutter Laboratories, Inc. | Pasteurized therapeutically active protein compositions |
| CN1137528A (en) * | 1995-10-31 | 1996-12-11 | 江西省博达生物工程研究所 | Production process for modified low temp. ethanolic human serum albumin |
| CN1406245A (en) * | 2000-10-24 | 2003-03-26 | 财团法人化学及血清疗法研究所 | Method of eliminating human serum albumin polymers |
| CN1717249A (en) * | 2002-11-25 | 2006-01-04 | 奥克塔法马股份有限公司 | Prekallikrein depleted plasma derived albumin fraction |
| CN103394076A (en) * | 2013-08-15 | 2013-11-20 | 江西博雅生物制药股份有限公司 | Process for preparing human serum albumin |
| CN104558156A (en) * | 2015-01-23 | 2015-04-29 | 郑州莱士血液制品有限公司 | Method for extracting human serum albumin from plasma and increasing yield |
-
2017
- 2017-07-13 CN CN201710568707.6A patent/CN107216383A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4440679A (en) * | 1980-03-05 | 1984-04-03 | Cutter Laboratories, Inc. | Pasteurized therapeutically active protein compositions |
| CN1137528A (en) * | 1995-10-31 | 1996-12-11 | 江西省博达生物工程研究所 | Production process for modified low temp. ethanolic human serum albumin |
| CN1406245A (en) * | 2000-10-24 | 2003-03-26 | 财团法人化学及血清疗法研究所 | Method of eliminating human serum albumin polymers |
| CN1717249A (en) * | 2002-11-25 | 2006-01-04 | 奥克塔法马股份有限公司 | Prekallikrein depleted plasma derived albumin fraction |
| CN103394076A (en) * | 2013-08-15 | 2013-11-20 | 江西博雅生物制药股份有限公司 | Process for preparing human serum albumin |
| CN104558156A (en) * | 2015-01-23 | 2015-04-29 | 郑州莱士血液制品有限公司 | Method for extracting human serum albumin from plasma and increasing yield |
Non-Patent Citations (2)
| Title |
|---|
| 方松: "FⅤ制作中醋酸溶液使用量对白蛋白回收率影响的观察", 《中国输血杂志》 * |
| 陈通威等: "降低人血白蛋白制品中PKA的方法初探", 《中国医药生物技术》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110317261A (en) * | 2018-03-28 | 2019-10-11 | 发贵科技（贵州）有限公司 | Improved two steps ultrafiltration produces human serum albumin |
| CN108864278A (en) * | 2018-07-27 | 2018-11-23 | 珠海宝锐生物科技有限公司 | A method of preparing molecular biosciences grade bovine serum albumin(BSA) |
| CN108864278B (en) * | 2018-07-27 | 2021-02-19 | 珠海宝锐生物科技有限公司 | Method for preparing molecular biological grade bovine serum albumin |
| CN110878120A (en) * | 2019-10-31 | 2020-03-13 | 华兰生物工程重庆有限公司 | Method for recovering albumin from component II supernatant |
| CN110878120B (en) * | 2019-10-31 | 2023-06-13 | 华兰生物工程重庆有限公司 | Method for recovering albumin from component II supernatant |
| CN116589559A (en) * | 2023-06-05 | 2023-08-15 | 广东丹霞生物制药有限公司 | Process for preparing human serum albumin |
| CN116589559B (en) * | 2023-06-05 | 2024-02-27 | 广东丹霞生物制药有限公司 | Process for preparing human serum albumin |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107216383A (en) | A kind of preparation method of human serum albumin and the human serum albumin | |
| CN103333240B (en) | Method for reclaiming human albumin from component IV precipitate | |
| CN102532304B (en) | Preparation method of human serum albumin | |
| CN101972479B (en) | Preparation process of intravenous injection human immunoglobulin | |
| CN102532307B (en) | Method for preparing human immunoglobulin | |
| CN103285135B (en) | Preparation technology for increasing clarity of poplar flower injection | |
| CN102161702A (en) | Method for producing human blood albumin | |
| CN104231075B (en) | Preparation process of human hepatitis B immunoglobulin | |
| CN104004084B (en) | A kind of production method of human albumin | |
| CN105254754A (en) | Preparation method for intravenous injection human immune globulin (PH4) | |
| CN104479011B (en) | A kind of preparation method of quiet note human normal immunoglobulin | |
| CN102816230B (en) | Preparation method of human serum albumin | |
| CN102311496B (en) | Method for recovering albumin from deposited components I+II+III from cold ethanol method | |
| CN116731162B (en) | Human immunoglobulin production process | |
| CN112521487A (en) | Improved production process of human serum albumin | |
| CN102212129B (en) | Method for extracting human fibrinogen from component I through column chromatography | |
| CN101161232A (en) | Method of producing intravenous injection human hepatitis b immune globulin | |
| CN103103170B (en) | Production process for cow or sheep hyaluronidase | |
| CN104193822B (en) | Process for preparing rabies human immune globulin | |
| CN113735963A (en) | Method for removing pigment in purification process of recombinant human serum albumin | |
| CN102816237A (en) | Preparation method of human anti-D immunoglobulin | |
| CN107383185A (en) | A kind of extracting method of high-purity bovine serum albumin | |
| CN104771713A (en) | Mulberry-ginger extract, and preparation process and application thereof | |
| CN112521486A (en) | Production method for separating human serum albumin from ethanol at low temperature under real-time control of ethanol | |
| CN105126100B (en) | IgM-rich human immunoglobulin preparation and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB03 | Change of inventor or designer information | ||
| CB03 | Change of inventor or designer information |
Inventor after: Mi Changming Inventor after: Yang Jingpeng Inventor after: Xu Xiaonan Inventor after: Xie Changfu Inventor after: Hu Huiheng Inventor before: Mi Changming Inventor before: Xie Changfu Inventor before: Hu Huiheng |
|
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170929 |