HRP20150639T1 - Postupak za proäśišä†avanje proteina - Google Patents
Postupak za proäśišä†avanje proteina Download PDFInfo
- Publication number
- HRP20150639T1 HRP20150639T1 HRP20150639TT HRP20150639T HRP20150639T1 HR P20150639 T1 HRP20150639 T1 HR P20150639T1 HR P20150639T T HRP20150639T T HR P20150639TT HR P20150639 T HRP20150639 T HR P20150639T HR P20150639 T1 HRP20150639 T1 HR P20150639T1
- Authority
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- Croatia
- Prior art keywords
- host cell
- recombinant
- recombinant protein
- seq
- extract
- Prior art date
Links
- 238000000034 method Methods 0.000 title claims 27
- 102000004169 proteins and genes Human genes 0.000 title 1
- 108090000623 proteins and genes Proteins 0.000 title 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 claims 13
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 claims 13
- 102000006010 Protein Disulfide-Isomerase Human genes 0.000 claims 6
- 108020003519 protein disulfide isomerase Proteins 0.000 claims 6
- 230000001580 bacterial effect Effects 0.000 claims 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims 2
- 108010016626 Dipeptides Proteins 0.000 claims 2
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 claims 2
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 claims 2
- 238000004587 chromatography analysis Methods 0.000 claims 2
- 239000012634 fragment Substances 0.000 claims 2
- 102000014914 Carrier Proteins Human genes 0.000 claims 1
- 241000588724 Escherichia coli Species 0.000 claims 1
- 101000611183 Homo sapiens Tumor necrosis factor Proteins 0.000 claims 1
- 229960000583 acetic acid Drugs 0.000 claims 1
- 108091008324 binding proteins Proteins 0.000 claims 1
- 238000005119 centrifugation Methods 0.000 claims 1
- 239000013604 expression vector Substances 0.000 claims 1
- 239000011536 extraction buffer Substances 0.000 claims 1
- 238000001914 filtration Methods 0.000 claims 1
- 239000012362 glacial acetic acid Substances 0.000 claims 1
- 238000010438 heat treatment Methods 0.000 claims 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims 1
- 102000057041 human TNF Human genes 0.000 claims 1
- 238000004255 ion exchange chromatography Methods 0.000 claims 1
- 238000000746 purification Methods 0.000 claims 1
- 238000000926 separation method Methods 0.000 claims 1
Classifications
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- 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
- C07K1/107—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides
-
- 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
- C07K1/18—Ion-exchange chromatography
-
- 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
- C07K1/22—Affinity chromatography or related techniques based upon selective absorption processes
-
- 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
- C07K1/30—Extraction; Separation; Purification by precipitation
-
- 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
- C07K1/36—Extraction; Separation; Purification by a combination of two or more processes of different types
-
- 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
-
- 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
- C07K14/475—Growth factors; Growth regulators
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/24—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
- C07K16/241—Tumor Necrosis Factors
-
- 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
- C12N9/90—Isomerases (5.)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/10—Immunoglobulins specific features characterized by their source of isolation or production
- C07K2317/14—Specific host cells or culture conditions, e.g. components, pH or temperature
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/55—Fab or Fab'
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Medicinal Chemistry (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- Analytical Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Gastroenterology & Hepatology (AREA)
- Toxicology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Enzymes And Modification Thereof (AREA)
- Peptides Or Proteins (AREA)
Claims (25)
1. Postupak za pročišćavanje rekombinantnog proteina iz gram-negativnog bakterijskog uzorka stanice domaćina ili njezinog ekstrakta pri čemu navedena stanica domaćina izražava rekombinantni protein i rekombinantnu disulfid izomerazu DsbC; pri čemu postupaka sadrži:
a) podešavanje pH uzorka stanice domaćina, ili njezinog ekstrakta do pH 5 ili manje da se istaloži rekombinantna disulfid izomeraza; i
b) odvajanje istaložene rekombinantne disulfid izomeraze iz rekombinantnog proteina da se proizvede uzorak rekombinantnog proteina.
2. Postupak prema patentnom zahtjevu 1, pri čemu je pH uzorka stanice domaćina ili njezinog ekstrakta podešen na pH od 4.5 ili manje.
3. Postupak prema patentnom zahtjevu 2, pri čemu je pH uzorka stanice domaćina ili njezinog ekstrakta podešen na pH od 4.5 do 3.0.
4. Postupak prema patentnom zahtjevu 2, pri čemu je pH uzorka stanice domaćina ili njezinog ekstrakta podešen na pH od 3 ili manje.
5. Postupak prema patentnom zahtjevu 4, pri čemu je u koraku a) dipeptid stanice domaćina vezujućeg proteina dalje istaložen i u koraku b) dipeptid stanice domaćina je dalje odvojen od rekombinantnog proteina.
6. Postupak prema bilo kojem od prethodnih patentnih zahtjeva, pri čemu disulfid izomeraza sadrži dodatak histidina.
7. Postupak prema bilo kojem od prethodnih patentnih zahtjeva, pri čemu je gram-negativna bakterijska stanica domaćina E. coli.
8. Postupak prema bilo kojem od prethodnih patentnih zahtjeva, pri čemu stanica domaćina sadrži Fkpa i/ili Skp.
9. Postupak prema bilo kojem od prethodnih patentnih zahtjeva, pri čemu u koraku a) pH uzorka stanice domaćina ili njezinog ekstrakta je podešen na pH od manje od 5 upotrebom ledene octene kiseline.
10. Postupak prema bilo kojem od prethodnih patentnih zahtjeva, pri čemu uzorak stanice domaćina ili njezinog ekstrakta se drži na pH od 5 ili manje tokom jednog sata ili manje.
11. Postupak prema bilo kojem od prethodnih patentnih zahtjeva, pri čemu separacija u koraku b) sadrži centrifugiranje i/ili filtriranje.
12. Postupak prema bilo kojem od prethodnih patentnih zahtjeva, pri čemu nakon koraka b) postupak sadrži dalji korak pročišćavanja uzorka rekombinantnog proteina kromatografijom.
13. Postupak prema patentnom zahtjevu 11, pri čemu je kromatografija kromatografija ionske izmjene.
14. Postupak prema bilo kojem od prethodnih patentnih zahtjeva, pri čemu nakon koraka b) pH uzorka rekombinantnog proteina je podešena na pH od 5 do 7.
15. Postupak prema bilo kojem od prethodnih patentnih zahtjeva, pri čemu pre koraka a) pufer za ekstrakciju je dodat uzorku stanice domaćina ili njegovom ekstraktu i uzorak stanice domaćina ili njegov ekstrakt je podvrgnut koraku termičke obrade.
16. Postupak prema bilo kojem od prethodnih patentnih zahtjeva, pri čemu rekombinantni protein ima pl od 6 do 9.
17. Postupak prema patentnom zahtjevu 16, pri čemu rekombinantni protein ima pl od 8 do 9.
18. Postupak prema bilo kojem od prethodnih patentnih zahtjeva, pri čemu rekombinantni protein je rekombinantno antitijelo.
19. Postupak prema patentnom zahtjevu 18, pri čemu je rekombinantno antitijelo monoklonalno, humanizirano ili himerno antitijelo, ili njihov fragment, kao što je njihov vezujući fragment.
20. Postupak prema patentnom zahtjevu 18 ili zahtjevu 19, pri čemu je rekombinantno antitijelo Fab ili Fab fragment.
21. Postupak prema bilo kojem od patentnih zahtjeva 18 do 20, pri čemu je rekombinantno antitijelo specifično za humani TNFɑ.
22. Postupak prema patentnom zahtjevu 21, pri čemu antitijelo sadrži teški lanac gdje varijabilna domena sadrži CDR koji ima sekvencu danu u SEQ ID NO:1 za CDRH1, SEQ ID NO:2, za CDRH2 i SEQ ID NO:3 za CDRH3 i laki lanac pri čemu varijabilna domena sadrži CDR koji ima sekvencu danu u SEQ ID NO:4 za CDRL1, SEQ ID NO:5 za CDRL2 i SEQ ID NO:6 za CDRL3.
23. Postupak prema patentnom zahtjevu 22, pri čemu antitijelo sadrži sekvencu varijabilne regije lakog lanca danu u SEQ ID NO: 7 i varijabilna regija teškog lanca je dana u SEQ ID NO: 8.
24. Postupak prema patentnom zahtjevu 23, pri čemu je antitijelo Fab fragment i sadrži teški lanac koji ima sekvencu danu u SEQ ID NO: 10 i laki lanac koji ima sekvencu danu u SEQ ID NO: 9.
25. Upotreba koraka podešavanja pH gram-negativnog bakterijskog uzorka stanice domaćina ili njezinog ekstrakta transformiranog ekspresijskim vektorom koji kodira rekombinantni protein na pH 5 ili manje kako bi se istaložila rekombinantna disulfid izomeraza stanice domaćina, odvojila istaložena rekombinantna disulfid izomeraza stanice domaćina iz rekombinantnog proteina i proizveo pročišćeni uzorak rekombinantnog proteina.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GBGB1012599.5A GB201012599D0 (en) | 2010-07-27 | 2010-07-27 | Process for purifying proteins |
| PCT/GB2011/001129 WO2012013930A2 (en) | 2010-07-27 | 2011-07-27 | Process for purifying proteins |
| EP11745810.9A EP2598516B8 (en) | 2010-07-27 | 2011-07-27 | Process for purifying proteins |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| HRP20150639T1 true HRP20150639T1 (hr) | 2015-07-17 |
Family
ID=42752868
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| HRP20150639TT HRP20150639T1 (hr) | 2010-07-27 | 2015-06-15 | Postupak za proäśišä†avanje proteina |
Country Status (24)
| Country | Link |
|---|---|
| US (1) | US9751930B2 (hr) |
| EP (1) | EP2598516B8 (hr) |
| JP (1) | JP5935222B2 (hr) |
| KR (1) | KR101944121B1 (hr) |
| CN (1) | CN103189385B (hr) |
| AU (1) | AU2011284548B2 (hr) |
| BR (1) | BR112013000177B1 (hr) |
| CA (1) | CA2804099C (hr) |
| CY (1) | CY1116526T1 (hr) |
| DK (1) | DK2598516T3 (hr) |
| ES (1) | ES2537877T3 (hr) |
| GB (1) | GB201012599D0 (hr) |
| HR (1) | HRP20150639T1 (hr) |
| HU (1) | HUE026049T2 (hr) |
| IL (1) | IL224044A (hr) |
| ME (1) | ME02162B (hr) |
| MX (1) | MX337184B (hr) |
| PL (1) | PL2598516T3 (hr) |
| PT (1) | PT2598516E (hr) |
| RS (1) | RS54085B1 (hr) |
| SG (2) | SG186974A1 (hr) |
| SI (1) | SI2598516T1 (hr) |
| SM (1) | SMT201500142B (hr) |
| WO (1) | WO2012013930A2 (hr) |
Families Citing this family (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP6132551B2 (ja) | 2009-09-24 | 2017-05-24 | ユセベ ファルマ ソシエテ アノニム | シャペロン活性を保持するプロテアーゼ欠損ＤｅｇＰ並びにノックアウトＴｓｐ及びｐｔｒ遺伝子を持つ、組換えタンパク質発現のための細菌株 |
| GB201000587D0 (en) | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial hoist strain |
| GB201000591D0 (en) | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial hoist strain |
| GB201000590D0 (en) | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial host strain |
| GB201001791D0 (en) | 2010-02-03 | 2010-03-24 | Ucb Pharma Sa | Process for obtaining antibodies |
| KR102023786B1 (ko) | 2011-07-13 | 2019-09-20 | 유씨비 파마, 에스.에이. | 재조합 dsbc를 발현하는 세균 숙주 균주 |
| GB201208367D0 (en) * | 2012-05-14 | 2012-06-27 | Ucb Pharma Sa | Biological product |
| SG11201407801VA (en) * | 2012-05-31 | 2014-12-30 | Agency Science Tech & Res | Methods for use of mixed multifunctional surfaces for reducing aggregate content in protein preparations |
| JP6297029B2 (ja) | 2012-05-31 | 2018-03-20 | エイジェンシー・フォー・サイエンス，テクノロジー・アンド・リサーチ | 多様な官能基を有する粒子による免疫グロブリンｇ調製物のクロマトグラフィー精製 |
| CA2966558C (en) * | 2014-11-05 | 2024-03-12 | Genentech, Inc. | Methods of producing two chain proteins in bacteria |
| EP3237432B1 (en) | 2014-12-22 | 2023-09-06 | UCB Biopharma SRL | Protein manufacture |
| US10626143B2 (en) | 2014-12-22 | 2020-04-21 | Ucb Biopharma Sprl | Method of protein manufacture |
| SG10202005917SA (en) * | 2015-03-06 | 2020-07-29 | Genentech Inc | Ultrapurified dsba and dsbc and methods of making and using the same |
| CN110272491B (zh) * | 2018-03-13 | 2023-01-24 | 江苏恒瑞医药股份有限公司 | 一种抗pd-1抗体的纯化工艺 |
| JP7259131B2 (ja) * | 2019-08-05 | 2023-04-17 | ワッカー ケミー アクチエンゲゼルシャフト | 発酵法で組換えタンパク質を放出する細菌株 |
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| JP5721951B2 (ja) | 2007-03-22 | 2015-05-20 | バイオジェン アイデック マサチューセッツ インコーポレイテッド | 抗体、抗体誘導体、および抗体断片を含む、ｃｄ１５４に特異的に結合する結合タンパク質ならびにその使用 |
| US7662587B1 (en) | 2009-03-05 | 2010-02-16 | E. I. Du Pont De Nemours And Company | Gene knockout mutations that increase peptide production |
| JP6132551B2 (ja) | 2009-09-24 | 2017-05-24 | ユセベ ファルマ ソシエテ アノニム | シャペロン活性を保持するプロテアーゼ欠損ＤｅｇＰ並びにノックアウトＴｓｐ及びｐｔｒ遺伝子を持つ、組換えタンパク質発現のための細菌株 |
| US8470552B2 (en) | 2009-10-12 | 2013-06-25 | Keck Graduate Institute | Strategy to reduce lactic acid production and control PH in animal cell culture |
| SI2496601T1 (sl) | 2009-11-05 | 2017-09-29 | F. Hoffmann-La Roche Ag | Tehnike in sestavek za sekrecijo heterolognih polipeptidov |
| GB201000587D0 (en) | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial hoist strain |
| GB201000588D0 (en) | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial host strain |
| GB201000590D0 (en) | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial host strain |
| GB201000591D0 (en) | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial hoist strain |
| GB201001791D0 (en) | 2010-02-03 | 2010-03-24 | Ucb Pharma Sa | Process for obtaining antibodies |
| EP2546267A1 (en) | 2011-07-13 | 2013-01-16 | UCB Pharma S.A. | Bacterial host strain expressing recombinant DsbC |
| KR102023786B1 (ko) | 2011-07-13 | 2019-09-20 | 유씨비 파마, 에스.에이. | 재조합 dsbc를 발현하는 세균 숙주 균주 |
| GB201208367D0 (en) | 2012-05-14 | 2012-06-27 | Ucb Pharma Sa | Biological product |
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2010
- 2010-07-27 GB GBGB1012599.5A patent/GB201012599D0/en not_active Ceased
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2011
- 2011-07-27 CN CN201180036417.8A patent/CN103189385B/zh active Active
- 2011-07-27 ES ES11745810.9T patent/ES2537877T3/es active Active
- 2011-07-27 PL PL11745810T patent/PL2598516T3/pl unknown
- 2011-07-27 WO PCT/GB2011/001129 patent/WO2012013930A2/en active Application Filing
- 2011-07-27 SI SI201130516T patent/SI2598516T1/sl unknown
- 2011-07-27 SG SG2013001318A patent/SG186974A1/en unknown
- 2011-07-27 SG SG2014005441A patent/SG196862A1/en unknown
- 2011-07-27 AU AU2011284548A patent/AU2011284548B2/en active Active
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