HRP20100053T1 - Postupak za dobivanje biološki aktivnog ljudskog g-csf iz inkluzijskih tijela - Google Patents
Postupak za dobivanje biološki aktivnog ljudskog g-csf iz inkluzijskih tijela Download PDFInfo
- Publication number
- HRP20100053T1 HRP20100053T1 HR20100053T HRP20100053T HRP20100053T1 HR P20100053 T1 HRP20100053 T1 HR P20100053T1 HR 20100053 T HR20100053 T HR 20100053T HR P20100053 T HRP20100053 T HR P20100053T HR P20100053 T1 HRP20100053 T1 HR P20100053T1
- Authority
- HR
- Croatia
- Prior art keywords
- csf
- concentration
- buffer
- inclusion bodies
- dissolution
- Prior art date
Links
- 238000000034 method Methods 0.000 title claims abstract 20
- 210000003000 inclusion body Anatomy 0.000 title claims abstract 7
- 108010024636 Glutathione Proteins 0.000 claims abstract 6
- 108010017080 Granulocyte Colony-Stimulating Factor Proteins 0.000 claims abstract 6
- 102000004269 Granulocyte Colony-Stimulating Factor Human genes 0.000 claims abstract 6
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 claims abstract 6
- 238000011210 chromatographic step Methods 0.000 claims abstract 4
- 108010053070 Glutathione Disulfide Proteins 0.000 claims abstract 3
- 239000003795 chemical substances by application Substances 0.000 claims abstract 3
- YPZRWBKMTBYPTK-BJDJZHNGSA-N glutathione disulfide Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@H](C(=O)NCC(O)=O)CSSC[C@@H](C(=O)NCC(O)=O)NC(=O)CC[C@H](N)C(O)=O YPZRWBKMTBYPTK-BJDJZHNGSA-N 0.000 claims abstract 3
- 101000746367 Homo sapiens Granulocyte colony-stimulating factor Proteins 0.000 claims abstract 2
- 238000007865 diluting Methods 0.000 claims abstract 2
- 238000004090 dissolution Methods 0.000 claims 6
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims 3
- 239000004202 carbamide Substances 0.000 claims 3
- 229960004198 guanidine Drugs 0.000 claims 3
- PJJJBBJSCAKJQF-UHFFFAOYSA-N guanidinium chloride Chemical group [Cl-].NC(N)=[NH2+] PJJJBBJSCAKJQF-UHFFFAOYSA-N 0.000 claims 3
- 238000005096 rolling process Methods 0.000 claims 3
- 239000000654 additive Substances 0.000 claims 2
- 230000000996 additive effect Effects 0.000 claims 2
- 238000005277 cation exchange chromatography Methods 0.000 claims 2
- 238000004587 chromatography analysis Methods 0.000 claims 2
- 150000002500 ions Chemical class 0.000 claims 2
- YPZRWBKMTBYPTK-UHFFFAOYSA-N oxidized gamma-L-glutamyl-L-cysteinylglycine Natural products OC(=O)C(N)CCC(=O)NC(C(=O)NCC(O)=O)CSSCC(C(=O)NCC(O)=O)NC(=O)CCC(N)C(O)=O YPZRWBKMTBYPTK-UHFFFAOYSA-N 0.000 claims 2
- 239000004475 Arginine Substances 0.000 claims 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 claims 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims 1
- 239000002738 chelating agent Substances 0.000 claims 1
- 238000010828 elution Methods 0.000 claims 1
- 238000001914 filtration Methods 0.000 claims 1
- 102000004169 proteins and genes Human genes 0.000 claims 1
- 108090000623 proteins and genes Proteins 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract 2
- 210000004027 cell Anatomy 0.000 abstract 1
- 239000000824 cytostatic agent Substances 0.000 abstract 1
- 230000001085 cytostatic effect Effects 0.000 abstract 1
- 230000004069 differentiation Effects 0.000 abstract 1
- 230000003394 haemopoietic effect Effects 0.000 abstract 1
- 229960001438 immunostimulant agent Drugs 0.000 abstract 1
- 239000003022 immunostimulating agent Substances 0.000 abstract 1
- 230000003308 immunostimulating effect Effects 0.000 abstract 1
- 230000010534 mechanism of action Effects 0.000 abstract 1
- 239000002243 precursor Substances 0.000 abstract 1
- 230000035755 proliferation Effects 0.000 abstract 1
- 230000003381 solubilizing effect Effects 0.000 abstract 1
- 230000004936 stimulating effect Effects 0.000 abstract 1
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
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/53—Colony-stimulating factor [CSF]
- C07K14/535—Granulocyte CSF; Granulocyte-macrophage CSF
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Toxicology (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Gastroenterology & Hepatology (AREA)
- Immunology (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Enzymes And Modification Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Steroid Compounds (AREA)
- Compounds Of Unknown Constitution (AREA)
Abstract
Postupak za dobivanje biološki aktivnog ljudskog G-CSF iz inkluzijskih tijela, naznačen time, da uključuje korake : a) otapanje G-CSF koji se nalazi u inkluzijskim tijelima pomoću pufera za otapanje koji sadrži denaturirani agens i reducirani glutation. b) ponovno motanje G-CSF razrjeđivanjem otopine sa puferom za ponovno motanje koji sadrži reducirane i oksidirane glutatione u ekvimolarnim količinama; ic) pročišćavanje ponovno zamotanog G-CSF barem jednim kromatografskim korakom. Patent sadrži još 17 patentnih zahtjeva.
Claims (18)
1. Postupak za dobivanje biološki aktivnog ljudskog G-CSF iz inkluzijskih tijela, naznačen time, da uključuje korake :
a) otapanje G-CSF koji se nalazi u inkluzijskim tijelima pomoću pufera za otapanje koji sadrži denaturirani agens i reducirani glutation.
b) ponovno motanje G-CSF razrjeđivanjem otopine sa puferom za ponovno motanje koji sadrži reducirane i oksidirane glutatione u ekvimolarnim količinama; i
c) pročišćavanje ponovno zamotanog G-CSF barem jednim kromatografskim korakom.
2. Postupak prema zahtjevu 1, naznačen time, da denaturirajući agens je gvanidin-HCI.
3. Postupak prema zahtjevu 2, naznačen time, da koncentracija gvanidin-HCI je 4.0 do 8.0 mol/L.
4. Postupak prema bilo kojem od prethodnih zahtjeva, naznačen time, da je koncentracija reduciranog glutationa u puferu za otapanje 10 do 200 mmol/L.
5. Postupak prema bilo kojem od prethodnih zahtjeva, naznačen time, da pufer za otapanje dodatno sadrži kelatni agens.
6. Postupak prema bilo kojem od prethodnih zahtjeva, naznačen time, da se koristi 4 do 20 mL pufera za otapanje po gramu inkluzijskih tijela.
7. Postupak prema bilo kojem od prethodnih zahtjeva, naznačen time, da pufer za otapanje nije odstranjen između otapanja i ponovnog motanja.
8. Postupak prema bilo kojem od prethodnih zahtjeva, naznačen time, da pufer za ponovno motanje sadrži aditiv koji je odabran iz skupine koja se sastoji od arginina, uree i gvanidin-HCI u koncentraciji koja pospješuje ponovno motanje.
9. Postupak prema patentnom zahtjevu 8, naznačen time, da je aditiv urea.
10. Postupak prema zahtjevu 8 ili 9, naznačen time, da pufer za ponovno motanje sadrži ureu u koncentraciji od 1.5 do 4.5 mol/L.
11. Postupak prema bilo kojem od prethodnih zahtjeva, naznačen time, da je koncentracija pojedinog reduciranog i oksidiranog glutationa 0.2 do 10 mmol/L.
12. Postupak prema bilo kojem od prethodnih zahtjeva, naznačena time, da je koncentracija proteina korištena za ponovno motanje ispod 2000 μg/mL.
13. Postupak prema bilo kojem od prethodnih zahtjeva, naznačen time, da se ponovno motanje odvija pri 4° C najmanje 3 sata.
14. Postupak prema bilo kojem od prethodnih zahtjeva, naznačen time, da je najmanje jedan kromatografski korak kromatografija ionskog izmjenjivača.
15. Postupak prema zahtjevu 14, naznačen time, da je kromatografija ionskog izmjenjivača kromatografija kationske izmjene.
16. Postupak prema zahtjevu 15, naznačen time, da se dio koji se ponovno mota zakiseljava prije kromatografije kationske izmjene.
17. Postupak prema zahtjevu 15 ili 16, naznačen time, da se eluacija G-CSF odvija pri pH vrijednost od 4.0 – 6.0.
18. Postupak prema bilo kojem od prethodnih zahtjeva, naznačen time, da se nakon ponovnog motanja, ali prije najmanje jednog kromatografskog koraka izvede filtriranje preko dubokog filtra.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE102004041639A DE102004041639A1 (de) | 2004-08-27 | 2004-08-27 | Verfahren zur Gewinnung von biologisch aktivem humanen G-CSF aus Inclusion Bodies |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| HRP20100053T1 true HRP20100053T1 (hr) | 2010-03-31 |
Family
ID=35406424
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| HR20100053T HRP20100053T1 (hr) | 2004-08-27 | 2010-01-29 | Postupak za dobivanje biološki aktivnog ljudskog g-csf iz inkluzijskih tijela |
Country Status (10)
| Country | Link |
|---|---|
| EP (1) | EP1630173B1 (hr) |
| AT (1) | ATE447584T1 (hr) |
| CY (1) | CY1109754T1 (hr) |
| DE (2) | DE102004041639A1 (hr) |
| DK (1) | DK1630173T3 (hr) |
| ES (1) | ES2336007T3 (hr) |
| HR (1) | HRP20100053T1 (hr) |
| PL (1) | PL1630173T3 (hr) |
| PT (1) | PT1630173E (hr) |
| SI (1) | SI1630173T1 (hr) |
Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE102005033250A1 (de) | 2005-07-15 | 2007-01-18 | Bioceuticals Arzneimittel Ag | Verfahren zur Reinigung von G-CSF |
| GB0605684D0 (en) * | 2006-03-21 | 2006-05-03 | Sicor Biotech Uab | Method For Purifying Granulocyte-Colony Stimulating Factor |
| WO2008096370A2 (en) * | 2007-02-05 | 2008-08-14 | Natco Pharma Limited | An efficient and novel purification method of recombinant hg-csf |
| MX2011013417A (es) | 2009-06-25 | 2012-03-29 | Amgen Inc | Procesos de purificacion por captura para proteinas expresadas en un sistema no mamifero. |
| CN102892780B (zh) | 2010-03-17 | 2015-07-29 | 通益制药有限公司 | 获得生物活性的重组人g-csf的方法 |
| CN101830977B (zh) * | 2010-05-07 | 2012-01-25 | 厦门特宝生物工程股份有限公司 | 一种重组人粒细胞刺激因子的纯化方法 |
| HUP1200172A2 (en) | 2012-03-19 | 2013-10-28 | Richter Gedeon Nyrt | Methods for refolding g-csf from inclusion bodies |
| HUP1200171A1 (hu) | 2012-03-19 | 2013-09-30 | Richter Gedeon Nyrt | Módszerek polipeptidek elõállítására |
| EP2978770B1 (en) | 2013-03-29 | 2019-11-06 | Dr. Reddy's Laboratories Limited | Refolding of proteins |
| WO2019175633A1 (en) * | 2018-03-15 | 2019-09-19 | Petiva Private Limited | Methods for refolding sucrose isomerase |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| IL86090A (en) * | 1987-04-16 | 1993-03-15 | Cetus Oncology Corp | Production of purified, biologically active, bacterially produced recombinant human csf-1 |
| DE4014750A1 (de) | 1990-05-08 | 1991-11-14 | Boehringer Mannheim Gmbh | Muteine des granulozyten-stimulierenden faktors (g-csf) |
| AU3350893A (en) * | 1991-07-29 | 1993-03-02 | Immunex Corporation | Process for isolating and purifying recombinant interleukin-7 |
| IL152804A0 (en) * | 2000-05-16 | 2003-06-24 | Bolder Biotechnology Inc | Methods for refolding proteins containing free cysteine residues |
| GB0019375D0 (en) * | 2000-08-07 | 2000-09-27 | Int Centre Genetic Eng & Bio | Method of polypeptide renaturation |
| WO2002057296A1 (en) * | 2001-01-16 | 2002-07-25 | Københavns Universitet | A method for refolding of proteins |
| JPWO2005033307A1 (ja) * | 2003-09-30 | 2006-12-14 | 技術研究組合 生物分子工学研究所 | 新規のリフォールディング方法およびその方法によって得られたタンパク質 |
-
2004
- 2004-08-27 DE DE102004041639A patent/DE102004041639A1/de not_active Ceased
-
2005
- 2005-08-29 SI SI200431323T patent/SI1630173T1/sl unknown
- 2005-08-29 DK DK05107881.4T patent/DK1630173T3/da active
- 2005-08-29 PT PT05107881T patent/PT1630173E/pt unknown
- 2005-08-29 PL PL05107881T patent/PL1630173T3/pl unknown
- 2005-08-29 AT AT05107881T patent/ATE447584T1/de not_active IP Right Cessation
- 2005-08-29 DE DE502005008432T patent/DE502005008432D1/de active Active
- 2005-08-29 EP EP05107881A patent/EP1630173B1/de not_active Revoked
- 2005-08-29 ES ES05107881T patent/ES2336007T3/es active Active
-
2010
- 2010-01-27 CY CY20101100082T patent/CY1109754T1/el unknown
- 2010-01-29 HR HR20100053T patent/HRP20100053T1/hr unknown
Also Published As
| Publication number | Publication date |
|---|---|
| EP1630173A3 (de) | 2006-03-08 |
| ATE447584T1 (de) | 2009-11-15 |
| EP1630173A2 (de) | 2006-03-01 |
| PL1630173T3 (pl) | 2010-04-30 |
| ES2336007T3 (es) | 2010-04-07 |
| PT1630173E (pt) | 2010-02-12 |
| CY1109754T1 (el) | 2014-09-10 |
| EP1630173B1 (de) | 2009-11-04 |
| SI1630173T1 (sl) | 2010-03-31 |
| DE102004041639A1 (de) | 2006-03-02 |
| DE502005008432D1 (de) | 2009-12-17 |
| DK1630173T3 (da) | 2010-03-15 |
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