JP5091127B2 - Il−18結合タンパク質の精製のための方法 - Google Patents
Il−18結合タンパク質の精製のための方法 Download PDFInfo
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- JP5091127B2 JP5091127B2 JP2008515220A JP2008515220A JP5091127B2 JP 5091127 B2 JP5091127 B2 JP 5091127B2 JP 2008515220 A JP2008515220 A JP 2008515220A JP 2008515220 A JP2008515220 A JP 2008515220A JP 5091127 B2 JP5091127 B2 JP 5091127B2
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- chromatography
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- protein
- affinity chromatography
- ligand
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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
- C07K1/22—Affinity chromatography or related techniques based upon selective absorption processes
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/281—Sorbents specially adapted for preparative, analytical or investigative chromatography
- B01J20/286—Phases chemically bonded to a substrate, e.g. to silica or to polymers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/30—Processes for preparing, regenerating, or reactivating
- B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
- B01J20/3231—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the coating or impregnating layer
- B01J20/3242—Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
- B01J20/3244—Non-macromolecular compounds
- B01J20/3246—Non-macromolecular compounds having a well defined chemical structure
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K9/00—Peptides having up to 20 amino acids, containing saccharide radicals and having a fully defined sequence; Derivatives thereof
- C07K9/006—Peptides having up to 20 amino acids, containing saccharide radicals and having a fully defined sequence; Derivatives thereof the peptide sequence being part of a ring structure
- C07K9/008—Peptides having up to 20 amino acids, containing saccharide radicals and having a fully defined sequence; Derivatives thereof the peptide sequence being part of a ring structure directly attached to a hetero atom of the saccharide radical, e.g. actaplanin, avoparcin, ristomycin, vancomycin
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
- B01D15/38—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving specific interaction not covered by one or more of groups B01D15/265 - B01D15/36
- B01D15/3804—Affinity chromatography
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Analytical Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Description
本発明は、タンパク質、特にIL−18結合タンパク質(IL−18BP)に対する精製方法の開発に基づく。
以下の段落は、本発明の化合物を構成する様々な化学部分の定義を提供し、明確に記載された定義がより広い定義を提供しない限り、明細書及び特許請求の範囲を通して一様に適用されることが意図される。
B1及びB2は、独立に、置換又は非置換C1−C6−アルキル、置換又は非置換C3−C8−シクロアルキル、及び置換又は非置換アリールから成る群から選択され、ここで、B1及びB2は、独立に、任意に、R2、OR2、SR2、又はN(R2)2で置換される;ここで、
各R2は、独立に、水素、置換又は非置換C1−C6−アルキル、置換又は非置換アリール、及び置換又は非置換C1−C6−アルキルアリールの群から選択される;
各Xは、独立に、窒素、水素を担持する炭素、塩素基を担持する炭素、又はシアノ基を担持する炭素を表し;且つ
Zは、マトリクスとの反応が可能な官能基である}。
各R2は、独立に、水素、C1−C6−アルキル、アリール、及びC1−C6−アルキルアリールから成る群から選択される。
(a)アフィニティークロマトグラフィーの溶出液を、固定化金属イオンアフィニティークロマトグラフィーにかける段階;
(b)金属イオンアフィニティークロマトグラフィーの溶出液を、疎水性電荷相互作用クロマトグラフィーにかける段階;
(c)疎水性電荷相互作用クロマトグラフィーの溶出液を、陽イオン交換クロマトグラフィーにかける段階;
(d)陽イオン交換クロマトグラフィーの流出物(flow−through)を、疎水性相互作用クロマトグラフィーにかける段階;
(e)疎水性相互作用の溶出液を、逆相クロマトグラフィーにかける段階。
a.疎水性相互作用クロマトグラフィー、
b.逆相クロマトグラフィー、及び
c.陰イオン交換クロマトグラフィー、
を含んで成る。
式(I)又は(II)の固定化アフィニティーリガンドを含んで成るアフィニティーカラムを、精製目的で用いた。
1.Elisa(導入物、流出物及び溶出液)
2.SDS−PAGE/CB(流出物及び溶出液)
3.SDS−PAGE/WB(流出物及び溶出液)
4.ブラッドフォード分析(導入物、流出物及び溶出液)
式(I)又は(II)の固定化アフィニティーリガンドを含んで成るアフィニティーカラムを、精製方法に用いた。
Claims (18)
- Zが−NH−である、請求項1に記載の使用。
- マトリクスがアガロースである、請求項1又は2に記載の使用。
- 流体からIL−18BPを捕獲するための、請求項1〜3のいずれか一項に記載のアフィニティーリガンドの使用。
- 請求項1〜4のいずれか一項に記載のアフィニティーリガンドを用いて、流体をアフィニティークロマトグラフィーにかけることを含んで成る、精製IL−18BPの生成のための方法。
- アフィニティークロマトグラフィーを室温で実施する、請求項5に記載の方法。
- IL−18BPの回収率が、アフィニティークロマトグラフィーの後で、40%超である、請求項5又は6に記載の方法。
- IL−18BPの回収率が、アフィニティークロマトグラフィーの後で、50%超である、請求項7に記載の方法。
- IL−18BPの純度が、アフィニティークロマトグラフィーの後で、60%超である、請求項5〜8のいずれか一項に記載の方法。
- IL−18BPの純度が、アフィニティークロマトグラフィーの後で、70%超である、請求項9に記載の方法。
- IL−18BPの純度が、アフィニティークロマトグラフィーの後で、80%超である、請求項9又は10に記載の方法。
- 単量体IL−18BPの比率が、アフィニティークロマトグラフィー段階の後で、約80%超である、請求項5〜11のいずれか一項に記載の方法。
- 疎水性相互作用クロマトグラフィー、逆相クロマトグラフィー及び陰イオン交換クロマトグラフィーから成る群から選択されるクロマトグラフィー段階を更に含んで成る、請求項5〜12のいずれか一項に記載の方法。
- 1つ以上のウイルス除去ろ過段階を更に含んで成る、請求項5又は6に記載の方法。
- IL−18BPがヒトの組換えIL−18BPである、請求項5〜14のいずれか一項に記載の方法。
- IL−18BPがヒトの組換えIL−18BPである、請求項1〜4のいずれか一項に記載の使用。
- 流体が無血清細胞培養上清である、請求項5〜14のいずれか一項に記載の方法。
- 流体が無血清細胞培養上清である、請求項1〜4のいずれか一項に記載の使用。
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP05105124 | 2005-06-10 | ||
EP05105124.1 | 2005-06-10 | ||
US69041405P | 2005-06-14 | 2005-06-14 | |
US60/690,414 | 2005-06-14 | ||
PCT/EP2006/063029 WO2006131550A1 (en) | 2005-06-10 | 2006-06-08 | Process for the purification of il-18 binding protein |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2008542432A JP2008542432A (ja) | 2008-11-27 |
JP5091127B2 true JP5091127B2 (ja) | 2012-12-05 |
Family
ID=34940147
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2008515220A Active JP5091127B2 (ja) | 2005-06-10 | 2006-06-08 | Il−18結合タンパク質の精製のための方法 |
Country Status (17)
Country | Link |
---|---|
US (1) | US8217152B2 (ja) |
EP (1) | EP1891088B1 (ja) |
JP (1) | JP5091127B2 (ja) |
AT (1) | ATE529433T1 (ja) |
AU (1) | AU2006256763B2 (ja) |
CA (1) | CA2610804C (ja) |
CY (1) | CY1112312T1 (ja) |
DK (1) | DK1891088T3 (ja) |
ES (1) | ES2375831T3 (ja) |
HR (1) | HRP20110948T1 (ja) |
IL (1) | IL187935A (ja) |
NO (1) | NO340957B1 (ja) |
PL (1) | PL1891088T3 (ja) |
PT (1) | PT1891088E (ja) |
RS (1) | RS52273B (ja) |
SI (1) | SI1891088T1 (ja) |
WO (1) | WO2006131550A1 (ja) |
Families Citing this family (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN100429315C (zh) | 2003-03-11 | 2008-10-29 | 雪兰诺实验室有限公司 | 含有mcmv ie2启动子的表达载体 |
JP5074030B2 (ja) * | 2003-05-13 | 2012-11-14 | メルク セローノ ソシエテ アノニム | Il−18結合タンパク質の活性変異体とその医学的応用 |
ATE484591T1 (de) | 2003-10-21 | 2010-10-15 | Merck Serono Sa | Minimale dna sequenz, die als chromatin-isolator wirkt, und deren verwendung für die protein- expression |
PT1689777E (pt) * | 2003-11-05 | 2007-05-31 | Ares Trading Sa | Processo para a purificação da proteína de ligação a il-18 |
RS50531B (sr) | 2004-03-01 | 2010-05-07 | Ares Trading S.A. | Upotreba podloge za ćelijsku kulturu bez sadržaja seruma za produkciju il-18bp u ćelijama sisara |
EP1761552B1 (en) * | 2004-06-29 | 2008-09-17 | Ares Trading S.A. | Process for the purification of il-18 binding protein |
GB0416699D0 (en) | 2004-07-27 | 2004-09-01 | Prometic Biosciences Ltd | Prion protein ligands and methods of use |
CA2609060C (en) | 2005-06-03 | 2014-07-15 | Urs Weber | Production of recombinant il-18 binding protein |
BRPI0611811A2 (pt) * | 2005-06-10 | 2008-12-09 | Prometic Biosciences Ltd | ligantes de ligaÇço À proteÍna |
DK2683736T3 (en) * | 2011-03-09 | 2018-04-23 | Cell Signaling Technology Inc | METHODS AND REAGENTS FOR CREATING MONOCLONAL ANTIBODIES |
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US4737462A (en) * | 1982-10-19 | 1988-04-12 | Cetus Corporation | Structural genes, plasmids and transformed cells for producing cysteine depleted muteins of interferon-β |
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US4959314A (en) * | 1984-11-09 | 1990-09-25 | Cetus Corporation | Cysteine-depleted muteins of biologically active proteins |
US5116943A (en) * | 1985-01-18 | 1992-05-26 | Cetus Corporation | Oxidation-resistant muteins of Il-2 and other protein |
US5017691A (en) * | 1986-07-03 | 1991-05-21 | Schering Corporation | Mammalian interleukin-4 |
US4879111A (en) * | 1986-04-17 | 1989-11-07 | Cetus Corporation | Treatment of infections with lymphokines |
US4965195A (en) * | 1987-10-26 | 1990-10-23 | Immunex Corp. | Interleukin-7 |
US4904584A (en) * | 1987-12-23 | 1990-02-27 | Genetics Institute, Inc. | Site-specific homogeneous modification of polypeptides |
DE3902948A1 (de) * | 1989-02-01 | 1990-08-09 | Telefunken Fernseh & Rundfunk | Verfahren zur uebertragung eines signals |
DE69230789T3 (de) | 1991-01-18 | 2007-10-31 | Amgen Inc., Thousand Oaks | Methoden zur behandlung von durch den tumor nekrose faktor ausgelösten krankheiten |
US6117996A (en) * | 1995-09-20 | 2000-09-12 | Novo Nordisk A/S | Triazine based ligands and use thereof |
IL121860A0 (en) * | 1997-08-14 | 1998-02-22 | Yeda Res & Dev | Interleukin-18 binding proteins their preparation and use |
US7220717B2 (en) * | 1997-08-14 | 2007-05-22 | Yeda Research And Development Company Ltd. | Interleukin-18 binding proteins, their preparation and use |
GB9910807D0 (en) * | 1999-05-10 | 1999-07-07 | Prometic Biosciences Limited | Novel detoxification agents and their use |
JP4944324B2 (ja) | 1999-07-13 | 2012-05-30 | ボルダー バイオテクノロジー, インコーポレイテッド | 免疫グロブリン融合タンパク質 |
IL131047A0 (en) | 1999-07-22 | 2001-01-28 | Yeda Res & Dev | Use of il-18 inhibitors |
PL206549B1 (pl) | 2000-02-21 | 2010-08-31 | Serono Lab | Zastosowanie inhibitora IL-18, wektora ekspresyjnego zawierającego kodującą go sekwencję, wektora wywołującego lub wzmacniającego endogenną produkcję inhibitora IL-18 oraz genetycznie zmodyfikowanej komórki |
DE60134009D1 (de) | 2000-05-05 | 2008-06-26 | Inst Nat Sante Rech Med | Verwendung von il-18 inhibitoren zur behandlung und/oder prävention von atherosklerose |
EE05534B1 (et) | 2001-01-29 | 2012-04-16 | Applied Research Systems Ars Holding N.V. | IL-18 inhibiitorite kasutamine ravimi valmistamiseks kardiomopaatia raviks ja/v?i ennetamiseks |
IL157800A0 (en) | 2001-03-08 | 2004-03-28 | Ares Trading Sa | Il-18 mutants, their production and use |
DK1425028T3 (da) | 2001-05-16 | 2010-03-01 | Yeda Res & Dev | Anvendelse af IL-18 inhibitorer til behandling eller forebyggelse af sepsis |
BR0210007A (pt) | 2001-05-25 | 2004-08-10 | Ares Trading Sa | Uso de inibidores de il-18 para tratamento ou prevenção de lesões do sistema nervoso central |
UA78516C2 (en) | 2001-08-10 | 2007-04-10 | Applied Research Systems | Use of inhibitors of il-18 for treatment and/or prevention of hypersensitivity disorders, and in particular of delayed-type hypersensitivity |
US9592267B2 (en) | 2002-03-22 | 2017-03-14 | Merck Serono Sa | Use of IL-18BP for treatment of peripheral vascular diseases |
CN100429315C (zh) * | 2003-03-11 | 2008-10-29 | 雪兰诺实验室有限公司 | 含有mcmv ie2启动子的表达载体 |
JP5074030B2 (ja) | 2003-05-13 | 2012-11-14 | メルク セローノ ソシエテ アノニム | Il−18結合タンパク質の活性変異体とその医学的応用 |
ATE484591T1 (de) * | 2003-10-21 | 2010-10-15 | Merck Serono Sa | Minimale dna sequenz, die als chromatin-isolator wirkt, und deren verwendung für die protein- expression |
PT1689777E (pt) * | 2003-11-05 | 2007-05-31 | Ares Trading Sa | Processo para a purificação da proteína de ligação a il-18 |
RS50531B (sr) | 2004-03-01 | 2010-05-07 | Ares Trading S.A. | Upotreba podloge za ćelijsku kulturu bez sadržaja seruma za produkciju il-18bp u ćelijama sisara |
EP1761552B1 (en) | 2004-06-29 | 2008-09-17 | Ares Trading S.A. | Process for the purification of il-18 binding protein |
CA2609060C (en) | 2005-06-03 | 2014-07-15 | Urs Weber | Production of recombinant il-18 binding protein |
BRPI0611811A2 (pt) * | 2005-06-10 | 2008-12-09 | Prometic Biosciences Ltd | ligantes de ligaÇço À proteÍna |
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2006
- 2006-06-08 JP JP2008515220A patent/JP5091127B2/ja active Active
- 2006-06-08 PL PL06763598T patent/PL1891088T3/pl unknown
- 2006-06-08 WO PCT/EP2006/063029 patent/WO2006131550A1/en active Application Filing
- 2006-06-08 RS RS20120013A patent/RS52273B/en unknown
- 2006-06-08 PT PT06763598T patent/PT1891088E/pt unknown
- 2006-06-08 AT AT06763598T patent/ATE529433T1/de active
- 2006-06-08 SI SI200631226T patent/SI1891088T1/sl unknown
- 2006-06-08 AU AU2006256763A patent/AU2006256763B2/en active Active
- 2006-06-08 CA CA2610804A patent/CA2610804C/en active Active
- 2006-06-08 US US11/916,087 patent/US8217152B2/en active Active
- 2006-06-08 ES ES06763598T patent/ES2375831T3/es active Active
- 2006-06-08 DK DK06763598.7T patent/DK1891088T3/da active
- 2006-06-08 EP EP06763598A patent/EP1891088B1/en active Active
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- 2007-12-06 IL IL187935A patent/IL187935A/en active IP Right Grant
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- 2008-01-08 NO NO20080131A patent/NO340957B1/no unknown
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2011
- 2011-12-17 HR HR20110948T patent/HRP20110948T1/hr unknown
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- 2012-01-18 CY CY20121100067T patent/CY1112312T1/el unknown
Also Published As
Publication number | Publication date |
---|---|
US20080200658A1 (en) | 2008-08-21 |
IL187935A (en) | 2015-10-29 |
CA2610804A1 (en) | 2006-12-14 |
WO2006131550A1 (en) | 2006-12-14 |
PT1891088E (pt) | 2011-12-19 |
CA2610804C (en) | 2013-11-19 |
CY1112312T1 (el) | 2015-12-09 |
ATE529433T1 (de) | 2011-11-15 |
ES2375831T3 (es) | 2012-03-06 |
SI1891088T1 (sl) | 2012-02-29 |
EP1891088A1 (en) | 2008-02-27 |
AU2006256763B2 (en) | 2012-08-02 |
PL1891088T3 (pl) | 2012-03-30 |
RS52273B (en) | 2012-10-31 |
US8217152B2 (en) | 2012-07-10 |
NO340957B1 (no) | 2017-07-31 |
EP1891088B1 (en) | 2011-10-19 |
JP2008542432A (ja) | 2008-11-27 |
AU2006256763A1 (en) | 2006-12-14 |
DK1891088T3 (da) | 2012-01-30 |
IL187935A0 (en) | 2008-03-20 |
HRP20110948T1 (hr) | 2012-01-31 |
NO20080131L (no) | 2008-03-07 |
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