JP5827955B2 - Peg化タンパク質の精製方法 - Google Patents
Peg化タンパク質の精製方法 Download PDFInfo
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- JP5827955B2 JP5827955B2 JP2012540414A JP2012540414A JP5827955B2 JP 5827955 B2 JP5827955 B2 JP 5827955B2 JP 2012540414 A JP2012540414 A JP 2012540414A JP 2012540414 A JP2012540414 A JP 2012540414A JP 5827955 B2 JP5827955 B2 JP 5827955B2
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- protein
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- pegylated
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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
- C07K1/14—Extraction; Separation; Purification
- C07K1/16—Extraction; Separation; Purification by chromatography
- C07K1/18—Ion-exchange chromatography
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/04—Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
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- 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)
- C12N9/644—Coagulation factor IXa (3.4.21.22)
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- 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)
- C12Y304/21022—Coagulation factor IXa (3.4.21.22)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
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- Biotechnology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
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- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
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- Chemical Kinetics & Catalysis (AREA)
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- Diabetes (AREA)
- Animal Behavior & Ethology (AREA)
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- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
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Description
PEG化FIXの精製
(a) 負荷(load)溶液
95%純度を超える純度のPEG化FIXをバッファー交換した。負荷(load)中のpH値は約6である。FIX-濃度は約1 mg/mlである。負荷(load)はおよそ5g産物/L樹脂
(b) カラム
Poros 50 HQ、15.7 mL。
(c) バッファー
- 平衡バッファー: 10 mM酢酸、90 mM酢酸ナトリウム、pH 〜 5.7
- 酸性洗浄: 5 CV酸性洗浄バッファー:酢酸ナトリウム65 mmol/kg、酢酸: 185 mmol/kg pH 〜 4.3; 5 CV
- 溶出液: 100%の平衡バッファーから100%の溶出バッファーまでの、5 CV線密度勾配(linear gradient) (10 mM ヒスチジン、50 mM NaCl、50 mM CaCl2)
- 再生液(Regeneration): 3 CV 1M NaCl
(d) 手順
工程 バッファー CV %-溶出バッファー
平衡 平衡バッファー 5 0%
適用
洗浄 平衡バッファー 3 0%
酸性洗浄 酸性洗浄バッファー 3 0%
(=洗浄2)
洗浄3 平衡バッファー 3 0%
溶出 溶出バッファー 5 0…100 %
再生 再生バッファー 3 0%
流速: 24 CV/h = 6.28 ml/分
温度: 5 ℃
実施態様1: 溶出バッファーを用いて陰イオン交換クロマトグラフィーを行う工程を含む、PEG化タンパク質の精製方法であって、前記クロマトグラフィーは、溶出の前に不純物除去工程を含むことを特徴とし、前記不純物除去工程は、酸性バッファーを用いて前記陰イオン交換カラムを洗浄する工程を含む、方法。
Claims (8)
- 溶出バッファーを用いて陰イオン交換クロマトグラフィーを行う工程を含む、PEG化タンパク質の精製方法であって、前記クロマトグラフィーは、溶出の前に不純物除去工程を含むことを特徴とし、前記不純物除去工程は、酸性バッファーを用いて前記陰イオン交換カラムを洗浄する工程を含み、前記不純物除去工程は、3.9から4.5の間のpHで酸性バッファーを用いて洗浄する工程を含む、方法。
- 前記不純物がST3Gal3である、請求項1に記載の方法。
- 酸性バッファーが、酢酸ナトリウムおよび/または酢酸を含む、請求項1または2に記載の方法。
- 平衡工程が酸性洗浄工程の前に実施される、請求項1から3のいずれか一項に記載の方法。
- 平衡工程が5から7のpHでの平衡バッファーの適用を含む、請求項4に記載の方法。
- 平衡バッファーが、酢酸および酢酸ナトリウムを含む、請求項4または5に記載の方法。
- PEG化タンパク質が、第II因子(FII), 第VII因子(FVII), 第IX因子(FIX), 第X因子(FX), プロテインSおよびプロテインCから選択される、請求項1から6のいずれか一項に記載の方法。
- 陰イオン交換物質がHQを含む、請求項1から7のいずれか一項に記載の方法。
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP09176881 | 2009-11-24 | ||
EP09176881.2 | 2009-11-24 | ||
US26448609P | 2009-11-25 | 2009-11-25 | |
US61/264,486 | 2009-11-25 | ||
PCT/EP2010/068112 WO2011064247A1 (en) | 2009-11-24 | 2010-11-24 | Method of purifying pegylated proteins |
Publications (3)
Publication Number | Publication Date |
---|---|
JP2013511566A JP2013511566A (ja) | 2013-04-04 |
JP2013511566A5 JP2013511566A5 (ja) | 2014-01-16 |
JP5827955B2 true JP5827955B2 (ja) | 2015-12-02 |
Family
ID=41401820
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2012540414A Active JP5827955B2 (ja) | 2009-11-24 | 2010-11-24 | Peg化タンパク質の精製方法 |
Country Status (5)
Country | Link |
---|---|
US (1) | US8697844B2 (ja) |
EP (1) | EP2504349B1 (ja) |
JP (1) | JP5827955B2 (ja) |
CN (2) | CN102639553A (ja) |
WO (1) | WO2011064247A1 (ja) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2019088608A1 (ko) * | 2017-10-30 | 2019-05-09 | 한국코러스 주식회사 | 콜로니자극인자와 폴리올이 접합된 접합물을 고수율로 제조하는 방법 |
Families Citing this family (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2532369B1 (en) | 2006-12-15 | 2017-11-01 | Baxalta GmbH | Factor VIIa-(poly)sialic acid conjugate having prolonged in vivo half-life |
ES2597954T3 (es) | 2009-07-27 | 2017-01-24 | Baxalta GmbH | Conjugados de proteína de la coagulación sanguínea |
US8642737B2 (en) | 2010-07-26 | 2014-02-04 | Baxter International Inc. | Nucleophilic catalysts for oxime linkage |
HUE028056T2 (en) | 2009-07-27 | 2016-11-28 | Baxalta GmbH | Blood coagulation protein conjugates |
US8809501B2 (en) | 2009-07-27 | 2014-08-19 | Baxter International Inc. | Nucleophilic catalysts for oxime linkage |
NZ597600A (en) | 2009-07-27 | 2014-05-30 | Lipoxen Technologies Ltd | Glycopolysialylation of non-blood coagulation proteins |
KR102025442B1 (ko) | 2010-12-22 | 2019-09-25 | 박스알타 인코퍼레이티드 | 단백질에 수용성 지방산 유도체를 접합하기 위한 물질 및 방법 |
EP3731871B1 (en) * | 2017-12-29 | 2023-10-04 | F. Hoffmann-La Roche AG | Process for providing pegylated protein composition |
CN111801120A (zh) | 2017-12-29 | 2020-10-20 | 豪夫迈·罗氏有限公司 | 用于提供聚乙二醇化蛋白质组合物的方法 |
JP7227633B2 (ja) * | 2017-12-29 | 2023-02-22 | エフ. ホフマン-ラ ロシュ エージー. | Peg化タンパク質組成物を提供するための方法 |
Family Cites Families (18)
Publication number | Priority date | Publication date | Assignee | Title |
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US5714583A (en) | 1995-06-07 | 1998-02-03 | Genetics Institute, Inc. | Factor IX purification methods |
WO1996040731A1 (en) | 1995-06-07 | 1996-12-19 | Mount Sinai School Of Medicine Of The City University Of New York | Pegylated modified proteins |
ES2198913T5 (es) * | 1998-05-06 | 2013-11-18 | Genentech, Inc. | Purificación de proteínas mediante cromatografía de intercambio iónico |
EP1084136B1 (en) | 1998-06-01 | 2004-08-25 | Genentech, Inc. | Separation of antibody monomers from its multimers by use of ion-exchange chromatography |
ES2564688T3 (es) | 2001-10-10 | 2016-03-28 | Ratiopharm Gmbh | Remodelación y glicoconjugación de poli(éter) a eritropoyetina |
US7265084B2 (en) | 2001-10-10 | 2007-09-04 | Neose Technologies, Inc. | Glycopegylation methods and proteins/peptides produced by the methods |
US7179617B2 (en) | 2001-10-10 | 2007-02-20 | Neose Technologies, Inc. | Factor IX: remolding and glycoconjugation of Factor IX |
US7795210B2 (en) | 2001-10-10 | 2010-09-14 | Novo Nordisk A/S | Protein remodeling methods and proteins/peptides produced by the methods |
US7473680B2 (en) | 2001-11-28 | 2009-01-06 | Neose Technologies, Inc. | Remodeling and glycoconjugation of peptides |
CN1703421B (zh) * | 2002-09-20 | 2010-06-23 | 法玛西亚公司 | 降低peg化蛋白的聚集体水平的方法 |
WO2006127896A2 (en) | 2005-05-25 | 2006-11-30 | Neose Technologies, Inc. | Glycopegylated factor ix |
EP2338333B1 (en) | 2003-04-09 | 2017-09-06 | ratiopharm GmbH | Glycopegylation methods and proteins/peptides produced by the methods |
US20060040856A1 (en) * | 2003-12-03 | 2006-02-23 | Neose Technologies, Inc. | Glycopegylated factor IX |
AU2004296860B2 (en) | 2003-12-03 | 2010-04-22 | Novo Nordisk A/S | Glycopegylated factor IX |
US20070105755A1 (en) * | 2005-10-26 | 2007-05-10 | Neose Technologies, Inc. | One pot desialylation and glycopegylation of therapeutic peptides |
AU2006280932A1 (en) * | 2005-08-19 | 2007-02-22 | Novo Nordisk A/S | Glycopegylated factor VII and factor Vila |
US8399632B2 (en) | 2007-02-28 | 2013-03-19 | Baxter International Inc. | Method for the purification of recombinant blood coagulation factor IX enriched in sulfated and/or phosphorylated molecules |
WO2008119815A1 (en) * | 2007-04-02 | 2008-10-09 | Novo Nordisk A/S | Subcutaneous administration of coagulation factor ix |
-
2010
- 2010-11-24 CN CN2010800532840A patent/CN102639553A/zh active Pending
- 2010-11-24 US US13/509,867 patent/US8697844B2/en active Active
- 2010-11-24 EP EP10784769.1A patent/EP2504349B1/en active Active
- 2010-11-24 CN CN201710713355.9A patent/CN107383158A/zh active Pending
- 2010-11-24 WO PCT/EP2010/068112 patent/WO2011064247A1/en active Application Filing
- 2010-11-24 JP JP2012540414A patent/JP5827955B2/ja active Active
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2019088608A1 (ko) * | 2017-10-30 | 2019-05-09 | 한국코러스 주식회사 | 콜로니자극인자와 폴리올이 접합된 접합물을 고수율로 제조하는 방법 |
KR20190047951A (ko) * | 2017-10-30 | 2019-05-09 | 한국코러스 주식회사 | 콜로니자극인자와 폴리올이 접합된 접합물을 고수율로 제조하는 방법 |
Also Published As
Publication number | Publication date |
---|---|
US20120244137A1 (en) | 2012-09-27 |
US8697844B2 (en) | 2014-04-15 |
WO2011064247A1 (en) | 2011-06-03 |
EP2504349B1 (en) | 2020-02-26 |
EP2504349A1 (en) | 2012-10-03 |
JP2013511566A (ja) | 2013-04-04 |
CN102639553A (zh) | 2012-08-15 |
CN107383158A (zh) | 2017-11-24 |
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