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WO2011024025A1 - An erythropoietin analogue and a method thereof - Google Patents

An erythropoietin analogue and a method thereof Download PDF

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Publication number
WO2011024025A1
WO2011024025A1 PCT/IB2009/007173 IB2009007173W WO2011024025A1 WO 2011024025 A1 WO2011024025 A1 WO 2011024025A1 IB 2009007173 W IB2009007173 W IB 2009007173W WO 2011024025 A1 WO2011024025 A1 WO 2011024025A1
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WIPO (PCT)
Prior art keywords
recombinant human
huepo
human erythropoietin
new molecular
comprised
Prior art date
Application number
PCT/IB2009/007173
Other languages
French (fr)
Inventor
Villoo Morawala Patell
Sunit Maity
Sudhir Joshi
Ashutosh Vyas
Gopal Dyaga
Srinivas Ireni
Original Assignee
Avesthagen Limited
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Publication date
Application filed by Avesthagen Limited filed Critical Avesthagen Limited
Publication of WO2011024025A1 publication Critical patent/WO2011024025A1/en

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    • 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
    • C07K14/505—Erythropoietin [EPO]
    • 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
    • A61P7/06—Antianaemics
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/90—Fusion polypeptide containing a motif for post-translational modification
    • C07K2319/91—Fusion polypeptide containing a motif for post-translational modification containing a motif for glycosylation

Definitions

  • the present invention relates generally to the use of novel fermentation and chromatographic procedures separately and jointly for the production of novel glycoprotein possessing one or more of the biological properties of naturally occurring erythropoietin, which has longer biological half-life compared to r- HuEPO. in biologically active form from fluids, especially mammalian host cell culture supematants.
  • Anemia is often an associated condition in patients with chronic kidney disease (CKD).
  • CKD chronic kidney disease
  • This anemia is a source of significant morbidity causing symptoms such as lack of energy, breathlessness, dizziness, angina, and poor appetite and decreased exercise tolerance.
  • the main cause of this anemia is a decreased production of erythropoietin, a naturally occurring hormone mainly produced by the kidney.
  • Much of the impaired quality of life and morbidity suffered by patients with CKD may be a consequence of this anemia and it may have a major impact on their sense of well-being as well as impairing their ability to work and affecting their social and sexual lives.
  • Recombinant human erythropoietin is currently available as a treatment for anaemia in end stage renal disease. Administration 2 to 3 times weekly is required in the majority of subjects.
  • the aim of inventing this new molecular analogue of recombinant human erythropoietin (r-HuEPO) is to obtain a therapeutic with a longer biological half-life compared to r-HuEPO, allowing a reduction of the frequency of injections necessary to maintain a desired level of systemic haemoglobin and haematocrit.
  • the chronic nature of CRF (unless a subject receives a kidney transplant) means that treatment may continue for a long part of the subject's life and multiple weekly injections of r-HuEPO can have a major impact on subjects and care givers.
  • the present invention relates generally to the use of novel chromatographic procedures separately and jointly for the production of novel glycoprotein possessing one or more of the biological properties of naturally occurring erythropoietin, in biologically active form from fluids, especially mammalian host cell culture supematants, a new molecular analogue of recombinant human erythropoietin, which has longer biological half-life compared to r-HuEPO.
  • the present invention relates to the use of novel fermentation process for the overexpression of novel glycoprotein possessing one or more of the biological properties of naturally occurring erythropoietin, new molecular analogue of recombinant human erythropoietin, which has longer biological half-life compared to r-HuEPO in CHO cells.
  • Summary of the present invention also includes use of novel chromatographic purification for rapid and efficient recovery of novel glycoprotein possessing one or more of the biological properties of naturally occurring erythropoietin, new molecular analogue of recombinant human erythropoietin, which has longer biological half-life compared to r-HuEPO.
  • the present invention provides an improved process for the cell culture manufacturing of new molecular analogue of recombinant human erythropoietin.
  • the invention provides system that helps in achieving proper glycosylation of new molecular analogue of recombinant human erythropoietin.
  • the invention also helps in maintaining higher cell viability for a longer period of time.
  • the cell culture manufacturing process starts with seeding the bioreactor at a predefined cell density in chemically defined medium.
  • the culture is fed in two stages, primary feeding which is designed to achieve the desired cell growth and, secondary feeding which is designed to maintain the higher cell viability and hyper glycosylation of the new molecular analogue of recombinant human erythropoietin.
  • the invention also relates to bioreactor operation procedure for the manufacturing of new molecular analogue of recombinant human erythropoietin.
  • This present invention relates to the rapid and efficient recovery of new molecular analogue of recombinant human erythropoietin from cell culture supernatant from Cell culture fluid by Ion exchange chromatography.
  • the first step of chromatography is an Anion exchanger in binding and elution mode. The elution is based on selective enrichment of Novel erythropoetin isoforms. Elution is carried out with Tris buffer containing 10OmM to 200 mM salt (pH 7-8). The set of isoforms eluted in this fraction contained the sialic acid residues approximately ranging from 14-18.
  • the differential glycosylation pattern in various isoforms formed determines the effective separation which in turn is affected by the salt concentration in the elution buffer.
  • the present invention specifically describes the separation of isoforms containing 14-18 sialic acid residues.
  • the additional glycosylation will affect the biochemical and biological properties of this novel erythropoietin analogue. Due to additional glycosylation, this novel analogue will have a slower serum clearance and therefore a longer half-life. The longer serum half-life will increase the in vivo biological activity and also will allow the analogue to be administered less frequently.
  • the Bioreactor Before seeding, the Bioreactor was assembled and sterilized by autoclaving at 121 0 C for 45 minutes. After sterilization, Bioreactor was charged with 7200ml of commercially available animal component free, chemically defined media. Afterwards, the bioreactor was kept under positive pressure with air at a flow rate of 0.2 Litre per minute. The bioreactor was aerated over night for 100% air saturation. The dO2 electrode was calibrated after stabilization of dissolved oxygen value. Sterile connection was created between the seed bottle and the seed port on the bioreactor head plate. The seed was then aseptically transferred to the bioreactor using peristaltic pump. The bioreactor was seeded with the density of 0.4 x 10 6 cells/mL After seeding, the bioreactor was allowed to run at following pre-set parameters:
  • the bioreactor was sampled at every 24/48 hours for in process quality control analysis.
  • the bioreactor process was a fed - batch process with feeding of different nutrients at definite culture stages.
  • the bioreactor was daily fed with 30OmL of primary feed that comprise of glucose, galactose, mannose, lipids, amino acids, vitamins, trace elements, cholesterol and growth factors.
  • the bioreactor was daily fed with 15OmL of secondary feed that comprise of Galactose, trace Elements, Manganese Chloride and Mannose.
  • the bioreactor was operated at following pre-set and controlled parameters;
  • the bioreactor was harvested at a cell viability of 70 - 80%.
  • the first step of chromatography is an Anion exchanger in binding and elution mode.
  • the elution is based on selective enrichment of Novel erythropoetin isoforms. Elution is carried out with Tris buffer containing 10OmM to 200 mM salt (pH 7-8).
  • the set of isoforms eluted in this fraction contained the sialic acid residues approximately ranging from 14-18 ( Figure 1)

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Public Health (AREA)
  • Hematology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Diabetes (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Animal Behavior & Ethology (AREA)
  • Engineering & Computer Science (AREA)
  • General Chemical & Material Sciences (AREA)
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Abstract

A process is provided comprising the use of novel fermentation and chromatographic procedures separately and jointly for the production of new molecular analogues of recombinant human erythropoietin (r'-HuEPO), which have a longer half-life compared to r-HuEPO, in biologically active form from fluids, especially mammalian host cell culture supernatants; wherein said analogues have a differential glycosylation pattern and contain sialic acid residues approximately ranging from 14-18. New molecular analogues of recombinant human erythropoietin r-HuEPO are also provided. •

Description

"AN ERYTHROPOIETIN ANALOGUE AND A METHOD THEREOF"
FIELD OF THE INVENTION
The present invention relates generally to the use of novel fermentation and chromatographic procedures separately and jointly for the production of novel glycoprotein possessing one or more of the biological properties of naturally occurring erythropoietin, which has longer biological half-life compared to r- HuEPO. in biologically active form from fluids, especially mammalian host cell culture supematants.
BACKGROUND AND PRIOR ART OF THE INVENTION
In past, various media and methods were used for the cell culture manufacturing of recombinant glycoprotein or monoclonal antibody. Commonly employed bioreactor process includes; batch, semi fed-batch, fed-batch, perfusion and continuous fermentation. The ever-increasing demand of monoclonal antibody and other recombinant proteins in properly glycosyalted forms have increased the prospects of cell culture process development. In addition the regulatory hurdles imposed on the serum containing process has led to the development of cell culture process in a completely chemically defined environment.
Numerous techniques have in the past been applied in preparative separations of biochemically significant materials. Commonly employed preparative separatory techniques include: ultrafiltration, column electrofocusing, flatbed electrofocusing, gel filtration, electrophoresis, isotachophoresis and various forms of chromatography. Among the commonly employed chromatoghraphic techniques are ion exchange and adsorption chromatography. The extensive application of recombinant methodologies to large-scale purification and production of eukaryotic protein has increased the prospect of obtaining the molecule in required quantity using simplified purification procedures.
Anemia is often an associated condition in patients with chronic kidney disease (CKD). There is a direct relationship between the severity of the anemia and the decline in renal function. This anemia is a source of significant morbidity causing symptoms such as lack of energy, breathlessness, dizziness, angina, and poor appetite and decreased exercise tolerance. The main cause of this anemia is a decreased production of erythropoietin, a naturally occurring hormone mainly produced by the kidney. Much of the impaired quality of life and morbidity suffered by patients with CKD may be a consequence of this anemia and it may have a major impact on their sense of well-being as well as impairing their ability to work and affecting their social and sexual lives. In the past, iron and folate were the main treatments for this condition and blood transfusions, with their associated risks of transmission of infection and induction of cytotoxic antibodies, which could jeopardize a future renal transplant, were used sparingly. In 1983 the cloning of the human gene for erythropoietin was achieved, production of recombinant human erythropoietin (r-HuEPO) followed and by 1986 the efficacy of r-HuEPO treatment in dialysis patients was first demonstrated.
Recombinant human erythropoietin is currently available as a treatment for anaemia in end stage renal disease. Administration 2 to 3 times weekly is required in the majority of subjects. The aim of inventing this new molecular analogue of recombinant human erythropoietin (r-HuEPO) is to obtain a therapeutic with a longer biological half-life compared to r-HuEPO, allowing a reduction of the frequency of injections necessary to maintain a desired level of systemic haemoglobin and haematocrit. The chronic nature of CRF (unless a subject receives a kidney transplant) means that treatment may continue for a long part of the subject's life and multiple weekly injections of r-HuEPO can have a major impact on subjects and care givers.
The development of Darbepoetin alfa arose from the understandings that the higher isoforms (those with a greater number of sialic acid residues) of recombinant human EPO were more potent biologically in vivo as a result of a longer circulating half-life than the lower isomers (those with a lower number of sialic acid residues). Because the majority of sialic acid residues are attached to the three N-linked glycosylation chains of the EPO molecule, attempts were made to synthesize EPO analogues with a greater number of N-linked carbohydrate chains. This was achieved using site- directed mutagenesis mutagenesis to change the amino acid sequence at sites not directly involved in binding to the EPO receptor. Thus, five- amino acid substitutions were implemented (allowing Darbepoetin alfa to carry a maximum of 22 sialic acid residues, compared with recombinant or endogenous EPO, which support a maximum of 14 sialic acid residues). The additional N-linked carbohydrate chains increased the molecular weight of epoetin from 30.4 to 37.1 kD, and the carbohydrate contribution to the molecule correspondingly increased from 40% to approximately 52%.
OBJECTIVES OF THE INVENTION
The present invention relates generally to the use of novel chromatographic procedures separately and jointly for the production of novel glycoprotein possessing one or more of the biological properties of naturally occurring erythropoietin, in biologically active form from fluids, especially mammalian host cell culture supematants, a new molecular analogue of recombinant human erythropoietin, which has longer biological half-life compared to r-HuEPO.
SUMMARY OF THE INVENTION
The present invention relates to the use of novel fermentation process for the overexpression of novel glycoprotein possessing one or more of the biological properties of naturally occurring erythropoietin, new molecular analogue of recombinant human erythropoietin, which has longer biological half-life compared to r-HuEPO in CHO cells.
Summary of the present invention also includes use of novel chromatographic purification for rapid and efficient recovery of novel glycoprotein possessing one or more of the biological properties of naturally occurring erythropoietin, new molecular analogue of recombinant human erythropoietin, which has longer biological half-life compared to r-HuEPO.
DETAILED DESCRIPTION OF THE INVENTION
The present invention provides an improved process for the cell culture manufacturing of new molecular analogue of recombinant human erythropoietin. In particular, the invention provides system that helps in achieving proper glycosylation of new molecular analogue of recombinant human erythropoietin. In addition, the invention also helps in maintaining higher cell viability for a longer period of time. The cell culture manufacturing process starts with seeding the bioreactor at a predefined cell density in chemically defined medium. The culture is fed in two stages, primary feeding which is designed to achieve the desired cell growth and, secondary feeding which is designed to maintain the higher cell viability and hyper glycosylation of the new molecular analogue of recombinant human erythropoietin. Furthermore the invention also relates to bioreactor operation procedure for the manufacturing of new molecular analogue of recombinant human erythropoietin.
This present invention relates to the rapid and efficient recovery of new molecular analogue of recombinant human erythropoietin from cell culture supernatant from Cell culture fluid by Ion exchange chromatography. The first step of chromatography is an Anion exchanger in binding and elution mode. The elution is based on selective enrichment of Novel erythropoetin isoforms. Elution is carried out with Tris buffer containing 10OmM to 200 mM salt (pH 7-8). The set of isoforms eluted in this fraction contained the sialic acid residues approximately ranging from 14-18. The differential glycosylation pattern in various isoforms formed determines the effective separation which in turn is affected by the salt concentration in the elution buffer. The present invention specifically describes the separation of isoforms containing 14-18 sialic acid residues. The additional glycosylation will affect the biochemical and biological properties of this novel erythropoietin analogue. Due to additional glycosylation, this novel analogue will have a slower serum clearance and therefore a longer half-life. The longer serum half-life will increase the in vivo biological activity and also will allow the analogue to be administered less frequently.
Example 1 :
Before seeding, the Bioreactor was assembled and sterilized by autoclaving at 1210C for 45 minutes. After sterilization, Bioreactor was charged with 7200ml of commercially available animal component free, chemically defined media. Afterwards, the bioreactor was kept under positive pressure with air at a flow rate of 0.2 Litre per minute. The bioreactor was aerated over night for 100% air saturation. The dO2 electrode was calibrated after stabilization of dissolved oxygen value. Sterile connection was created between the seed bottle and the seed port on the bioreactor head plate. The seed was then aseptically transferred to the bioreactor using peristaltic pump. The bioreactor was seeded with the density of 0.4 x 106 cells/mL After seeding, the bioreactor was allowed to run at following pre-set parameters:
pH: 6.9 - 7.2
dO2: 30 - 60% of air saturation
Temperature: 30 - 38°C.
Stir speed: 70-80RPM The bioreactor was sampled at every 24/48 hours for in process quality control analysis. The bioreactor process was a fed - batch process with feeding of different nutrients at definite culture stages. During first 72 hrs of culture time, the bioreactor was daily fed with 30OmL of primary feed that comprise of glucose, galactose, mannose, lipids, amino acids, vitamins, trace elements, cholesterol and growth factors. Starting from 96 hrs of culture age the bioreactor was daily fed with 15OmL of secondary feed that comprise of Galactose, trace Elements, Manganese Chloride and Mannose. During first 120 hrs of culture age the bioreactor was operated at following pre-set and controlled parameters;
pH: 7.1 ±0.1
dO2: 30 - 60% of air saturation
Temperature: 36 - 37°C.
Stir speed: 70-80RPM
From 120 to 288 hrs the bioreactor was operated at following parameters; pH: 6.8 ±0.05
dO2: 30 - 60% of air saturation
Temperature: 32 + 0.5 0C.
Stir speed: 70-80RPM
The bioreactor was harvested at a cell viability of 70 - 80%.
Example 2:
The first step of chromatography is an Anion exchanger in binding and elution mode. The elution is based on selective enrichment of Novel erythropoetin isoforms. Elution is carried out with Tris buffer containing 10OmM to 200 mM salt (pH 7-8). The set of isoforms eluted in this fraction contained the sialic acid residues approximately ranging from 14-18 (Figure 1)

Claims

We claim:
1. A process for recovering a new molecular analogue of recombinant human erythropoietin, which has longer biological half-life compared to r-HuEPO. a) subjecting to anion exchange chromatography to obtain the new molecular analogue of recombinant human erythropoietin, which has longer biological half-life compared to r-HuEPO
2. The process as claimed in claim 1 , wherein said supernatant is mammalian host cell culture supernatant.
3. The process as claimed in claim 1 , wherein said supernatant is cell culture derived fluid.
4. The process as claimed in claim 1 , wherein said supernatant is a mammalian cell culture derived fluid.
5. The process as claimed in claim 1 , wherein said culture supematant(s) are clarified before contacting resins.
6. A cell culture manufacturing process for the manufacturing of recombinant a new molecular analogue of recombinant human erythropoietin, which has longer biological half-life compared to r-HuEPO.
7. A process of increasing protein glycosylation using a predefined secondary feed.
8. The process as claimed in claim 11 , where secondary feed can be comprised of Carbohydrates.
9. The process as claimed in claim 11 , where secondary feed can be comprised of Carbohydrates and Trace Elements.
10. The process as claimed in claim 11 , where secondary feed can be comprised of Carbohydrates, Trace Elements and Manganese Chloride.
11. The process as claimed in claim 11 , where secondary feed can be comprised of Galactose and Mannose
12. The process as claimed in claim 11, where secondary feed can be comprised of 1-2 Molar Galactose and Mannose
13. The process as claimed in claim 11 , where secondary feed can be comprised of 25-75% of 1-2 Molar Galactose and Mannose
14. The process as claimed in claim 11, where secondary feed can be comprised of 25-75% of 1-2 Molar Galactose and Mannose along with Trace Elements
15. The process as claimed in claim 11 , where secondary feed can be comprised of 25-75% of 1-2 Molar Galactose and Mannose along with 0.01 - 1.0% Trace Elements and Manganese Chloride.
16. The process as claimed in claim 11 , where secondary feed can be comprised of 25-75% of 1-2 Molar Galactose and Mannose along with 0.01 - 1.0% Trace Elements and 0.02 - 5 micro molar Manganese Chloride.
17. A new molecular analogue of recombinant human erythropoietin.
18. A new molecular analogue of recombinant human erythropoietin containing more glycosylation sites compared to r-HuEPO.
19. As in claim 18, additional glycosylation site is an N-linked glycosylation site having an N-linked carbohydrate chain attached thereto.
20. As in claim 18, additional glycosylation site is an O-linked glycosylation site having an O-linked carbohydrate chain attached thereto.
21. Combination of both as mentioned in claim 19 and 20.
22. A new molecular analogue of recombinant human erythropoietin, which has longer biological half-life compared to r-HuEPO.
23. A new molecular analogue of recombinant human erythropoietin, which has longer biological half-life compared to r-HuEPO containing Sialic acid residues more than 14.
24. A new molecular analogue of recombinant human erythropoietin, which has longer biological half-life compared to r-HuEPO containing Sialic acid residues less than 18.
25. A new molecular analogue of recombinant human erythropoietin, which has longer biological half-life compared to r-HuEPO containing Sialic acid residues between 14-18.
26. A new molecular analogue of recombinant human erythropoietin will have in vivo biological activity of increasing haematocrit.
27. A new molecular analogue of recombinant human erythropoietin will have more in vivo biological activity of increasing haematocrit in comparison to r- HuEPO.
28. A new molecular analogue of recombinant human erythropoietin which has delayed serum clearance than r-HuEPO.
PCT/IB2009/007173 2009-08-28 2009-10-21 An erythropoietin analogue and a method thereof WO2011024025A1 (en)

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8895709B2 (en) 2008-10-20 2014-11-25 Abbvie Inc. Isolation and purification of antibodies using protein A affinity chromatography
US8906646B2 (en) 2006-09-13 2014-12-09 Abbvie Inc. Fed-batch method of making human anti-TNF-alpha antibody
US8911964B2 (en) 2006-09-13 2014-12-16 Abbvie Inc. Fed-batch method of making human anti-TNF-alpha antibody
US8921526B2 (en) 2013-03-14 2014-12-30 Abbvie, Inc. Mutated anti-TNFα antibodies and methods of their use
US8946395B1 (en) 2013-10-18 2015-02-03 Abbvie Inc. Purification of proteins using hydrophobic interaction chromatography
US9017687B1 (en) 2013-10-18 2015-04-28 Abbvie, Inc. Low acidic species compositions and methods for producing and using the same using displacement chromatography
US9062106B2 (en) 2011-04-27 2015-06-23 Abbvie Inc. Methods for controlling the galactosylation profile of recombinantly-expressed proteins
US9067990B2 (en) 2013-03-14 2015-06-30 Abbvie, Inc. Protein purification using displacement chromatography
US9085618B2 (en) 2013-10-18 2015-07-21 Abbvie, Inc. Low acidic species compositions and methods for producing and using the same
US9109010B2 (en) 2008-10-20 2015-08-18 Abbvie Inc. Viral inactivation during purification of antibodies cross reference to related applications
US9150645B2 (en) 2012-04-20 2015-10-06 Abbvie, Inc. Cell culture methods to reduce acidic species
US9181337B2 (en) 2013-10-18 2015-11-10 Abbvie, Inc. Modulated lysine variant species compositions and methods for producing and using the same
US9181572B2 (en) 2012-04-20 2015-11-10 Abbvie, Inc. Methods to modulate lysine variant distribution
US9193787B2 (en) 2012-04-20 2015-11-24 Abbvie Inc. Human antibodies that bind human TNF-alpha and methods of preparing the same
US9206390B2 (en) 2012-09-02 2015-12-08 Abbvie, Inc. Methods to control protein heterogeneity
US9234033B2 (en) 2012-09-02 2016-01-12 Abbvie, Inc. Methods to control protein heterogeneity
US9249182B2 (en) 2012-05-24 2016-02-02 Abbvie, Inc. Purification of antibodies using hydrophobic interaction chromatography
US9499614B2 (en) 2013-03-14 2016-11-22 Abbvie Inc. Methods for modulating protein glycosylation profiles of recombinant protein therapeutics using monosaccharides and oligosaccharides
US9550826B2 (en) 2013-11-15 2017-01-24 Abbvie Inc. Glycoengineered binding protein compositions
CN106498098A (en) * 2016-12-13 2017-03-15 镇江东方生物工程设备技术有限责任公司 A kind of animal cell culture Satellite tank DO100% calibration steps
US9598667B2 (en) 2013-10-04 2017-03-21 Abbvie Inc. Use of metal ions for modulation of protein glycosylation profiles of recombinant proteins

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0640619B2 (en) * 1993-08-17 2005-03-23 Kirin-Amgen, Inc. Erythropoietin analogs with additional glycosylation sites
US20070161084A1 (en) * 2005-12-08 2007-07-12 Amgen Inc. Production of glycoproteins using manganese
EP1064951B1 (en) * 1999-07-02 2007-08-22 F. Hoffmann-La Roche Ag Erythropoietin derivatives
WO2007136752A2 (en) * 2006-05-19 2007-11-29 Glycofi, Inc. Erythropoietin compositions

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0640619B2 (en) * 1993-08-17 2005-03-23 Kirin-Amgen, Inc. Erythropoietin analogs with additional glycosylation sites
EP1064951B1 (en) * 1999-07-02 2007-08-22 F. Hoffmann-La Roche Ag Erythropoietin derivatives
US20070161084A1 (en) * 2005-12-08 2007-07-12 Amgen Inc. Production of glycoproteins using manganese
WO2007136752A2 (en) * 2006-05-19 2007-11-29 Glycofi, Inc. Erythropoietin compositions

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US9090867B2 (en) 2006-09-13 2015-07-28 Abbvie Inc. Fed-batch method of making anti-TNF-alpha antibody
US8906646B2 (en) 2006-09-13 2014-12-09 Abbvie Inc. Fed-batch method of making human anti-TNF-alpha antibody
US8911964B2 (en) 2006-09-13 2014-12-16 Abbvie Inc. Fed-batch method of making human anti-TNF-alpha antibody
US10119118B2 (en) 2006-09-13 2018-11-06 Abbvie Inc. Modified serum-free cell culture medium
US9284371B2 (en) 2006-09-13 2016-03-15 Abbvie Inc. Methods of producing adalimumab
US9234032B2 (en) 2006-09-13 2016-01-12 Abbvie Inc. Fed-batch methods for producing adalimumab
US9073988B2 (en) 2006-09-13 2015-07-07 Abbvie Inc. Fed batch method of making anti-TNF-alpha antibodies
US9018361B2 (en) 2008-10-20 2015-04-28 Abbvie Inc. Isolation and purification of antibodies using protein a affinity chromatography
US8895709B2 (en) 2008-10-20 2014-11-25 Abbvie Inc. Isolation and purification of antibodies using protein A affinity chromatography
US9109010B2 (en) 2008-10-20 2015-08-18 Abbvie Inc. Viral inactivation during purification of antibodies cross reference to related applications
US9255143B2 (en) 2011-04-27 2016-02-09 Abbvie Inc. Methods for controlling the galactosylation profile of recombinantly-expressed proteins
US9505834B2 (en) 2011-04-27 2016-11-29 Abbvie Inc. Methods for controlling the galactosylation profile of recombinantly-expressed proteins
US9365645B1 (en) 2011-04-27 2016-06-14 Abbvie, Inc. Methods for controlling the galactosylation profile of recombinantly-expressed proteins
US9062106B2 (en) 2011-04-27 2015-06-23 Abbvie Inc. Methods for controlling the galactosylation profile of recombinantly-expressed proteins
US9090688B2 (en) 2011-04-27 2015-07-28 Abbvie Inc. Methods for controlling the galactosylation profile of recombinantly-expressed proteins
US9150645B2 (en) 2012-04-20 2015-10-06 Abbvie, Inc. Cell culture methods to reduce acidic species
US9181572B2 (en) 2012-04-20 2015-11-10 Abbvie, Inc. Methods to modulate lysine variant distribution
US9193787B2 (en) 2012-04-20 2015-11-24 Abbvie Inc. Human antibodies that bind human TNF-alpha and methods of preparing the same
US9683033B2 (en) 2012-04-20 2017-06-20 Abbvie, Inc. Cell culture methods to reduce acidic species
US9505833B2 (en) 2012-04-20 2016-11-29 Abbvie Inc. Human antibodies that bind human TNF-alpha and methods of preparing the same
US9334319B2 (en) 2012-04-20 2016-05-10 Abbvie Inc. Low acidic species compositions
US9708400B2 (en) 2012-04-20 2017-07-18 Abbvie, Inc. Methods to modulate lysine variant distribution
US9957318B2 (en) 2012-04-20 2018-05-01 Abbvie Inc. Protein purification methods to reduce acidic species
US9359434B2 (en) 2012-04-20 2016-06-07 Abbvie, Inc. Cell culture methods to reduce acidic species
US9346879B2 (en) 2012-04-20 2016-05-24 Abbvie Inc. Protein purification methods to reduce acidic species
US9249182B2 (en) 2012-05-24 2016-02-02 Abbvie, Inc. Purification of antibodies using hydrophobic interaction chromatography
US9206390B2 (en) 2012-09-02 2015-12-08 Abbvie, Inc. Methods to control protein heterogeneity
US9512214B2 (en) 2012-09-02 2016-12-06 Abbvie, Inc. Methods to control protein heterogeneity
US9290568B2 (en) 2012-09-02 2016-03-22 Abbvie, Inc. Methods to control protein heterogeneity
US9234033B2 (en) 2012-09-02 2016-01-12 Abbvie, Inc. Methods to control protein heterogeneity
US8921526B2 (en) 2013-03-14 2014-12-30 Abbvie, Inc. Mutated anti-TNFα antibodies and methods of their use
US9708399B2 (en) 2013-03-14 2017-07-18 Abbvie, Inc. Protein purification using displacement chromatography
US9499614B2 (en) 2013-03-14 2016-11-22 Abbvie Inc. Methods for modulating protein glycosylation profiles of recombinant protein therapeutics using monosaccharides and oligosaccharides
US9067990B2 (en) 2013-03-14 2015-06-30 Abbvie, Inc. Protein purification using displacement chromatography
US9598667B2 (en) 2013-10-04 2017-03-21 Abbvie Inc. Use of metal ions for modulation of protein glycosylation profiles of recombinant proteins
US9522953B2 (en) 2013-10-18 2016-12-20 Abbvie, Inc. Low acidic species compositions and methods for producing and using the same
US9200070B2 (en) 2013-10-18 2015-12-01 Abbvie, Inc. Low acidic species compositions and methods for producing and using the same
US9315574B2 (en) 2013-10-18 2016-04-19 Abbvie, Inc. Low acidic species compositions and methods for producing and using the same
US9266949B2 (en) 2013-10-18 2016-02-23 Abbvie, Inc. Low acidic species compositions and methods for producing and using the same
US9200069B2 (en) 2013-10-18 2015-12-01 Abbvie, Inc. Low acidic species compositions and methods for producing and using the same
US9181337B2 (en) 2013-10-18 2015-11-10 Abbvie, Inc. Modulated lysine variant species compositions and methods for producing and using the same
US9688752B2 (en) 2013-10-18 2017-06-27 Abbvie Inc. Low acidic species compositions and methods for producing and using the same using displacement chromatography
US9499616B2 (en) 2013-10-18 2016-11-22 Abbvie Inc. Modulated lysine variant species compositions and methods for producing and using the same
US8946395B1 (en) 2013-10-18 2015-02-03 Abbvie Inc. Purification of proteins using hydrophobic interaction chromatography
US9085618B2 (en) 2013-10-18 2015-07-21 Abbvie, Inc. Low acidic species compositions and methods for producing and using the same
US9017687B1 (en) 2013-10-18 2015-04-28 Abbvie, Inc. Low acidic species compositions and methods for producing and using the same using displacement chromatography
US9550826B2 (en) 2013-11-15 2017-01-24 Abbvie Inc. Glycoengineered binding protein compositions
CN106498098A (en) * 2016-12-13 2017-03-15 镇江东方生物工程设备技术有限责任公司 A kind of animal cell culture Satellite tank DO100% calibration steps

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