CN100432222C - 一种重组人尿激酶原的纯化方法 - Google Patents
一种重组人尿激酶原的纯化方法 Download PDFInfo
- Publication number
- CN100432222C CN100432222C CNB2004100188356A CN200410018835A CN100432222C CN 100432222 C CN100432222 C CN 100432222C CN B2004100188356 A CNB2004100188356 A CN B2004100188356A CN 200410018835 A CN200410018835 A CN 200410018835A CN 100432222 C CN100432222 C CN 100432222C
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- liquid
- chromatographic column
- buffered saline
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- Expired - Lifetime
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- 108090000435 Urokinase-type plasminogen activator Proteins 0.000 title claims abstract description 67
- 238000000746 purification Methods 0.000 title claims abstract description 34
- 102000003990 Urokinase-type plasminogen activator Human genes 0.000 title claims abstract description 33
- 229960005356 urokinase Drugs 0.000 title claims abstract description 33
- 102000010911 Enzyme Precursors Human genes 0.000 title claims abstract description 31
- 108010062466 Enzyme Precursors Proteins 0.000 title claims abstract description 31
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 34
- 238000004113 cell culture Methods 0.000 claims abstract description 21
- 238000000034 method Methods 0.000 claims abstract description 21
- 229920002684 Sepharose Polymers 0.000 claims abstract description 16
- 239000012506 Sephacryl® Substances 0.000 claims abstract description 15
- 229920002271 DEAE-Sepharose Polymers 0.000 claims abstract description 14
- 238000001042 affinity chromatography Methods 0.000 claims abstract description 7
- 238000005571 anion exchange chromatography Methods 0.000 claims abstract description 4
- 238000005227 gel permeation chromatography Methods 0.000 claims abstract description 4
- 239000000872 buffer Substances 0.000 claims description 74
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 claims description 64
- 239000002953 phosphate buffered saline Substances 0.000 claims description 64
- 239000007788 liquid Substances 0.000 claims description 63
- 238000010521 absorption reaction Methods 0.000 claims description 47
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 claims description 37
- 230000002572 peristaltic effect Effects 0.000 claims description 37
- 102000004169 proteins and genes Human genes 0.000 claims description 28
- 238000012546 transfer Methods 0.000 claims description 22
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 21
- 239000012530 fluid Substances 0.000 claims description 20
- 239000000523 sample Substances 0.000 claims description 20
- 239000012228 culture supernatant Substances 0.000 claims description 19
- 239000006228 supernatant Substances 0.000 claims description 16
- 239000000243 solution Substances 0.000 claims description 13
- 239000000047 product Substances 0.000 claims description 11
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims description 10
- 239000007983 Tris buffer Substances 0.000 claims description 10
- 238000005349 anion exchange Methods 0.000 claims description 10
- 238000013016 damping Methods 0.000 claims description 10
- 238000010828 elution Methods 0.000 claims description 10
- 238000011010 flushing procedure Methods 0.000 claims description 10
- 238000012544 monitoring process Methods 0.000 claims description 10
- 238000002953 preparative HPLC Methods 0.000 claims description 10
- 238000001179 sorption measurement Methods 0.000 claims description 10
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims description 10
- 238000000825 ultraviolet detection Methods 0.000 claims description 10
- 239000012470 diluted sample Substances 0.000 claims description 7
- 125000002091 cationic group Chemical group 0.000 claims description 4
- 230000002485 urinary effect Effects 0.000 claims description 2
- 241000124008 Mammalia Species 0.000 abstract description 3
- 238000004587 chromatography analysis Methods 0.000 abstract description 3
- 210000004102 animal cell Anatomy 0.000 abstract description 2
- 238000005341 cation exchange Methods 0.000 abstract description 2
- 238000011084 recovery Methods 0.000 abstract description 2
- LGKDEBYYRWXEDL-UHFFFAOYSA-N 2-aminobenzenecarboximidamide Chemical compound NC(=N)C1=CC=CC=C1N LGKDEBYYRWXEDL-UHFFFAOYSA-N 0.000 abstract 1
- 102100031358 Urokinase-type plasminogen activator Human genes 0.000 description 29
- 210000004027 cell Anatomy 0.000 description 16
- 238000012545 processing Methods 0.000 description 11
- 108010073863 saruplase Proteins 0.000 description 10
- 238000004519 manufacturing process Methods 0.000 description 5
- 239000013612 plasmid Substances 0.000 description 5
- WPANETAWYGDRLL-UHFFFAOYSA-N 4-aminobenzenecarboximidamide Chemical compound NC(=N)C1=CC=C(N)C=C1 WPANETAWYGDRLL-UHFFFAOYSA-N 0.000 description 4
- 230000003321 amplification Effects 0.000 description 4
- 238000005277 cation exchange chromatography Methods 0.000 description 4
- 239000002299 complementary DNA Substances 0.000 description 4
- 238000003199 nucleic acid amplification method Methods 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 239000012634 fragment Substances 0.000 description 3
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 description 2
- 101150074155 DHFR gene Proteins 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 239000002356 single layer Substances 0.000 description 2
- 238000001890 transfection Methods 0.000 description 2
- 240000008791 Antiaris toxicaria Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 244000270834 Myristica fragrans Species 0.000 description 1
- 235000009421 Myristica fragrans Nutrition 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- 108010022394 Threonine synthase Proteins 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 238000005377 adsorption chromatography Methods 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 125000002057 carboxymethyl group Chemical group [H]OC(=O)C([H])([H])[*] 0.000 description 1
- 238000000975 co-precipitation Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 239000003527 fibrinolytic agent Substances 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000001702 nutmeg Substances 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 238000005096 rolling process Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 229960000103 thrombolytic agent Drugs 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 230000003245 working effect Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
- C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
- C12N9/6424—Serine endopeptidases (3.4.21)
- C12N9/6456—Plasminogen activators
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/16—Extraction; Separation; Purification by chromatography
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
- C12Y304/21—Serine endopeptidases (3.4.21)
- C12Y304/21073—Serine endopeptidases (3.4.21) u-Plasminogen activator (3.4.21.73), i.e. urokinase
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- General Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
Description
Claims (7)
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CNB2004100188356A CN100432222C (zh) | 2004-04-05 | 2004-04-05 | 一种重组人尿激酶原的纯化方法 |
PCT/CN2005/000259 WO2005111207A1 (fr) | 2004-04-05 | 2005-03-03 | Procede de purification de prourokinase |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CNB2004100188356A CN100432222C (zh) | 2004-04-05 | 2004-04-05 | 一种重组人尿激酶原的纯化方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN1680550A CN1680550A (zh) | 2005-10-12 |
CN100432222C true CN100432222C (zh) | 2008-11-12 |
Family
ID=35067308
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CNB2004100188356A Expired - Lifetime CN100432222C (zh) | 2004-04-05 | 2004-04-05 | 一种重组人尿激酶原的纯化方法 |
Country Status (2)
Country | Link |
---|---|
CN (1) | CN100432222C (zh) |
WO (1) | WO2005111207A1 (zh) |
Families Citing this family (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101880655B (zh) * | 2009-05-06 | 2013-12-04 | 中国人民解放军军事医学科学院生物工程研究所 | 一种纯化cho细胞表达尿激酶原的方法 |
CN103717284B (zh) * | 2011-06-08 | 2016-02-24 | 新加坡科技研究局 | 通过约束共水合色谱纯化生物制品 |
CN103159849B (zh) * | 2011-12-08 | 2015-05-13 | 鲁南新时代生物技术有限公司 | 一种制备重组人胰岛素原的方法 |
CN103789291B (zh) * | 2014-02-24 | 2016-08-17 | 东北制药集团股份有限公司 | 一种重组大肠杆菌发酵液中分离纯化重组人尿激酶原的制备工艺 |
CN106867984B (zh) * | 2015-12-11 | 2021-11-12 | 天士力生物医药股份有限公司 | 一种重组人尿激酶原的纯化方法 |
CN106867985A (zh) * | 2015-12-11 | 2017-06-20 | 上海天士力药业有限公司 | 一种重组人尿激酶原的纯化及去除病毒方法 |
CN106867983A (zh) * | 2015-12-11 | 2017-06-20 | 上海天士力药业有限公司 | 一种重组人尿激酶原的病毒去除方法 |
CN108226363B (zh) * | 2017-12-04 | 2019-01-18 | 赵欣雨 | 一种利用hplc检测尿激酶分子组分比的方法 |
CN114191385B (zh) * | 2021-12-22 | 2023-06-30 | 天士力生物医药股份有限公司 | 一种重组人尿激酶原注射液及其制备方法 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH03164172A (ja) * | 1989-11-21 | 1991-07-16 | Tosoh Corp | ウロキナーゼ前駆体様蛋白質の精製方法 |
JPH0471488A (ja) * | 1990-07-13 | 1992-03-06 | Tosoh Corp | ウロキナーゼ前駆体の精製法 |
CN1164536A (zh) * | 1996-05-03 | 1997-11-12 | 中国人民解放军军事医学科学院生物工程研究所 | 溶血栓新药-重组人尿激酶原的纯化工艺 |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR100454891B1 (ko) * | 2001-06-08 | 2004-11-09 | 씨제이 주식회사 | 활성형 재조합 인체 프로유로키나제의 정제 방법 |
-
2004
- 2004-04-05 CN CNB2004100188356A patent/CN100432222C/zh not_active Expired - Lifetime
-
2005
- 2005-03-03 WO PCT/CN2005/000259 patent/WO2005111207A1/zh active Application Filing
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH03164172A (ja) * | 1989-11-21 | 1991-07-16 | Tosoh Corp | ウロキナーゼ前駆体様蛋白質の精製方法 |
JPH0471488A (ja) * | 1990-07-13 | 1992-03-06 | Tosoh Corp | ウロキナーゼ前駆体の精製法 |
CN1164536A (zh) * | 1996-05-03 | 1997-11-12 | 中国人民解放军军事医学科学院生物工程研究所 | 溶血栓新药-重组人尿激酶原的纯化工艺 |
Also Published As
Publication number | Publication date |
---|---|
WO2005111207A1 (fr) | 2005-11-24 |
CN1680550A (zh) | 2005-10-12 |
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Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
ASS | Succession or assignment of patent right |
Owner name: SHANGHAI DAYS TASLY PHARMACEUTICAL CO., LTD. Free format text: FORMER OWNER: TIANJIN TASLY PHARMACEUTICAL CO., LTD. Free format text: FORMER OWNER: SHANGHAI DAYS TASLY PHARMACEUTICAL CO., LTD. Effective date: 20050902 |
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C41 | Transfer of patent application or patent right or utility model | ||
TA01 | Transfer of patent application right |
Effective date of registration: 20050902 Address after: No. 280 Curie Road, Zhangjiang hi tech park, Shanghai Applicant after: SHANGHAI TASLY PHARMACEUTICAL Co.,Ltd. Address before: White road, Beichen science and Technology Park in Tianjin City Xinyi Liaohe Road No. 1 Applicant before: TIANJIN TASLY PHARMACEUTICAL Co.,Ltd. Co-applicant before: SHANGHAI TASLY PHARMACEUTICAL Co.,Ltd. |
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C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
C41 | Transfer of patent application or patent right or utility model | ||
TR01 | Transfer of patent right |
Effective date of registration: 20160304 Address after: 300410 Tianjin City Hedong District of Beichen Puji Road No. 2 city of the modern Chinese Medicine Patentee after: TASLY PHARMACEUTICAL GROUP Co.,Ltd. Address before: 201203, Curie road 280, Zhangjiang hi tech park, Shanghai Patentee before: SHANGHAI TASLY PHARMACEUTICAL Co.,Ltd. |
|
CP03 | Change of name, title or address |
Address after: 300410 Tianjin city Beichen District Huaihe road and road intersection Dingjiang tianzhijiao Park forensic Center for Intellectual Property Department Patentee after: TASLY PHARMACEUTICAL GROUP Co.,Ltd. Address before: 300410 Tianjin City Hedong District of Beichen Puji Road No. 2 city of the modern Chinese Medicine Patentee before: TASLY PHARMACEUTICAL GROUP Co.,Ltd. |
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CP03 | Change of name, title or address | ||
TR01 | Transfer of patent right |
Effective date of registration: 20180301 Address after: 201203 China (Shanghai) pilot free trade zone Curie road 280, 1, 2 Patentee after: SHANGHAI TASLY PHARMACEUTICAL Co.,Ltd. Address before: 300410 Tianjin city Beichen District Huaihe road and road intersection Dingjiang tianzhijiao Park forensic Center for Intellectual Property Department Patentee before: TASLY PHARMACEUTICAL GROUP Co.,Ltd. |
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TR01 | Transfer of patent right | ||
CP03 | Change of name, title or address |
Address after: 201203 No. 1 and No. 2 Curie Road, China (Shanghai) Free Trade Pilot Area, Shanghai Patentee after: TASLY PHARMACEUTICAL GROUP Co.,Ltd. Address before: China (Shanghai) free trade pilot area 280 Curie Road 1, 2 Patentee before: SHANGHAI TASLY PHARMACEUTICAL Co.,Ltd. |
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CP03 | Change of name, title or address | ||
PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: A purification method of recombinant human prourokinase Effective date of registration: 20210629 Granted publication date: 20081112 Pledgee: Bank of Shanghai Limited by Share Ltd. Pudong branch Pledgor: TASLY PHARMACEUTICAL GROUP Co.,Ltd. Registration number: Y2021310000043 |
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PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
CX01 | Expiry of patent term | ||
CX01 | Expiry of patent term |
Granted publication date: 20081112 |