WO2011015919A1 - A highly efficient process of purification and production of recombinant infliximab - Google Patents
A highly efficient process of purification and production of recombinant infliximab Download PDFInfo
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- WO2011015919A1 WO2011015919A1 PCT/IB2010/001896 IB2010001896W WO2011015919A1 WO 2011015919 A1 WO2011015919 A1 WO 2011015919A1 IB 2010001896 W IB2010001896 W IB 2010001896W WO 2011015919 A1 WO2011015919 A1 WO 2011015919A1
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- supernatant
- monoclonal antibody
- recovery
- buffer
- infliximab
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- 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
Definitions
- the present invention relates generally to the use of novel fermentation and chromatographic procedures separately and jointly for the production of Recombinant Infliximab, a monoclonal antibody to human TNF-alpha, in biologically active form from fluids, especially mammalian host cell culture supernatants.
- the invention relates primarily to a new way of recovering higher yield of purified Recombinant Infliximab by using bio analytical techniques such as but not limited to chromatographic procedures.
- Infliximab is a recombinant chimeric (partially humanized) murine monoclonal antibody glycoprotein that binds specifically to human tumor necrosis factor alpha (TNF-alpha) with an association constant of 1010 M-I.
- TNF-alpha is a naturally occurring cytokine that is involved in normal inflammatory and immune responses. Elevated concentrations of TNF-alpha are known to exist in the joints of RA (Rheumatoid Arthritis) patients and the stools of Crohn's disease patients.
- RA Ratoid Arthritis
- Infliximab reduces the infiltration of inflammatory cells into inflamed areas of the joints and also reduces expression of molecules mediating cellular adhesion, chemo attraction and tissue degradation e.g.
- Infliximab reduces infiltration of inflammatory cells and decreases TNF-alpha production in inflamed areas of the intestine.
- Infliximab in combination with methotrexate, is indicated for the reduction in signs and symptoms of RA in patients who have had an inadequate response to methotrexate. Infliximab is also indicated for the reduction in signs and symptoms of Crohn's disease in patients with moderately to severely active Crohn's disease who have had an inadequate response to conventional therapy.
- Infliximab can be used to treat multiple health ailments and hence it is required to be produced on a commercial large scale set up.
- Challenges in the purification of proteins viz. monoclonal antibodies (MAbs) include reducing production cost, developing robust processes for both product purity and viral clearance, and integrating upstream and downstream processes.
- Purifying MAbs for therapeutic use requires highly selective and robust technologies to achieve the very high purity required for biopharmaceuticals. Processing such valuable product streams at large-scale while manufacturing to such high standards requires careful technology design and execution. This is because supernatant containing the target MAbs are most often obtained from the culture of live cells and are variable in both product content and composition.
- the lowest production cost is achieved not by increased process complexity but by the opposite: Operating faster and with the fewest and most efficient downstream steps reduces investments in validation and subsequent batch failures (Kelley B. Very large scale monoclonal antibody purification: the case for conventional unit operations. Biotechnology Programme. 2007; 23(5):995-1008).
- the present invention constitutes a pathbreaking improvement in purification and recovery of Recombinant Infliximab.
- the main object of the present invention is to use novel fermentation and chromatographic procedures for rapid and efficient recovery of Recombinant Infliximab, a monoclonal antibody to human TNF-alpha, from cell culture supernatant.
- This invention is directed to a process for facilitating maximum product (recombinant Infliximab) recovery.
- the present invention relates to the use of novel fermentation process for the over expression of Recombinant Infliximab, a monoclonal antibody to human TNF-alpha, protein in CHO cells.
- the present invention also relates to the use of novel chromatographic procedures separately and jointly for the production of Recombinant Infliximab, a monoclonal antibody to human
- TNF-alpha protein, in biologically active form from fluids, especially mammalian host cell culture supernatants.
- Figure 1 Process chromatogram of affinity chromatography: Chromatographic profile of the first step of recovery of the target protein from the affinity chromatography. This is the purification chromatogram from a supernatant containing Infliximab as explained in example 1 (Supernatant derived from a harvested media containing Infliximab after clarification). The blue line represents the UV absorbance at 280nm. Majority of the other protein comes out in the flow through as can be observed in the figure.
- FIG. 2 Process chromatogram of anion chromatography: Chromatographic profile of the second step of recovery of the target protein from the anion exchange chromatography.
- This figure shows a chromatogram of purification from the eluent of the affinity chromatography as explained in the example 1 (Start material on to this column was the buffer exchanged neutralized elute containing the infliximab from the affinity column). It is clear from the figure that the majority of the load has come out in the flow through mode. A minor peak in the end may be some amount of the inflixmab has bound, as it is clear from the recovery that the majority of it has come out in follow-through.
- FIG. 3 Process chromatogram of cation chromatography: Chromatographic profile of the final step of recovery of the target protein from the cation exchange chromatography, This is a chromatogram from step 3 of a process according to the invention and it is more specifically a polishing step as explained in the example 1. The extra peak at the end is strongly bound protein that comes off with time.
- Fig 4 Electrophoretic pattern of Drug substance: showing comparable molecular weight with RMP where Lane No. 1 : Molecular weight Marker, Lane No. 2 : RMP and Lane No. 3 : Formulated Drug Substance. Electrophoretic profile reveals the purity and identity of the target protein with the Originator. The electrophoretic diagram depicts the stability of the drug substance and also the purity of the process close to homogeneity and comparability of the drug substance with that of the Originator.
- Fig 5 Protein A HPLC profile of Drug substance showing comparable with RMP: This profile shows similar retention time in the protein A column. This is one of the orthogonal methods for estimation of target protein (recombinant inflximab).
- This figure is an analytical protein A column chromatogrm of the purified drug substance containing inflximab as it was compared with that of the originator.
- the extra peak seen at the beginning of the chromatograms is from the buffer as it can be observed in both the chromatogram (innovators as well as the inflximab purified product).
- This method not only gives the protein estimation but also the purity of the product with respect to the RMP ⁇ Reference Medicinal Product - Remicade®)
- the present invention relates to a process for the recovery of Recombinant Monoclonal antibody to TNF alpha from cell culture supernatant arising out of cell culture fluid by means of an innovative series of chromatographic steps.
- the present invention also relates to the recombinant Monoclonal antibody to TNF alpha purification procedure involving serial application different chromatographic techniques as mentioned previously. AU different steps, conditions and compositions are disclosed in the invention Culture supernatants are clarified before chromatographic treatment.
- First affinity chromatography is used for capture of recombinant Monoclonal antibody to TNF alpha. This process comprises selective binding of the desired compound to specific affinity resin and then elution with elution buffer.
- Resultant monoclonal antibody to TNF alpha containing eluent fractions are still enriched with biologically active material. So they are further subjected to processing by Ion exchange chromatographic step. Anion exchange and cation exchange chromatography are used for removal of process related impurities like host cell protein and host cell DNA.
- the multi-modal chromatography capture step followed by ion exchange chromatography steps yield a product that increases its step recovery from 94.46% in Step 1 to 98.65% in Step 3. [Table I].
- the cell culture harvest is clarified by using a cellulose disposable filter.
- This filter is expressly customized with the effective filtration area being 650-1000 cm 2 and an operating pressure of up to 30 psi.
- the filtrate is checked for turbidity and target protein content.
- Affinity chromatography is used in binding and elution mode with column of 32 mm diameter for capturing; with Tris buffer pH 7.2 - 7.6 as equilibration buffer. After the sample is loaded on to the column, it is washed with equilibration buffer followed by 50 mM Tris-Cl. Then the second wash is performed with 50 mM Tris-Cl containing 100-400 mM NaCl pH 7-8 buffer solution.
- the protein of interest is eluted with citrate buffer pH 3.0-3.8 (Fig 1).
- the eluate is held for 45 - 60 min at acidic pH at room temperature for virus inactivation and later neutralized.
- the Protein A eluate fraction is taken for next purification step.
- Anion exchange chromatography in negative binding mode is then carried out at an operational flow rate of 140 cm/hr.
- the column has been equilibrated with Tris buffer pH 6.8 - 7.2.
- Protein of interest is collected in flow through. This step is used for the removal of process related impurities like leachate protein A, host cell DNA and host cell protein. (Fig 2). Thereafter, the flow through is filtered for virus removal using viral removal filter having an effective filtration area of 0.01 m 2 .
- the filtrate is then buffer exchanged using a 50 kDa TFF membrane.
- the buffer used for the diafiltration process is Tris buffer ph 6.8-7.2.
- Cation exchange chromatography is carried out with the diafiltered protein solution after equilibrating the column with Tris buffer pH 6.8-7.2.
- the protein of interest is eluted with elution buffer using NaCl salt gradient. This step is used for the removal of process related impurities like host cell DNA and host cell protein, which are there in traces (Fig 3).
- the eluate is buffer exchanged and concentrated using a 50 kDa TFF membrane at a Trans Membrane Pressure (TMP) of 5 - 10 psi.
- TMP Trans Membrane Pressure
- the drug substance was characterized as per the specifications.
- the Drug Substance (Active Pharmaceutical Ingredient) is formulated using formulation buffer pH 7.2, containing following constituents contains, 500 mg sucrose, 0.5 mg polysorbate 80, 2.2 mg monobasic sodium phosphate, monohydrate, and 6.1 mg dibasic sodium phosphate, dihydrate yielding a concentration of 10mg/ml of Infliximab. No preservatives are present. The efficiency of the process is depicted in terms of the recovery per say of the target protein. (Table 1).
- Table 1 Depicts the percentage of recovery of the target protein in purification process.
- the formulated material is characterized as per the specifications set by product development specification so as to meet the physico-chemical parameter comparable with that of the originator.
- a 10% SDS PAGE under reducing condition is studied for the sample derived from the PAGE showed a clear corresponding band with RMP (Reference Medicinal Product -Remicade®) reflecting the fact that there is a comparable purity and integrity (Fig 4).
- RMP Reference Medicinal Product -Remicade®
- Protein A HPLC profile carried out during this step showed in test molecule, which was very much similar to the RMP. This technique is used as one of the orthogonal method for estimation of target protein (Fig 5).
Abstract
The present invention relates to the use of novel fermentation and chromatographic procedures separately and jointly for the production of recombinant Monoclonal antibody to TNF alpha, in biologically active form from fluids, especially mammalian host cell culture supernatants. The recovery via the novel procedures can be very useful in downstream processing in commercial scale.
Description
A Highly Efficient Process of Purification and Production of
Recombinant Infliximab
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 Recombinant Infliximab, a monoclonal antibody to human TNF-alpha, in biologically active form from fluids, especially mammalian host cell culture supernatants. The invention relates primarily to a new way of recovering higher yield of purified Recombinant Infliximab by using bio analytical techniques such as but not limited to chromatographic procedures.
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.
Infliximab is a recombinant chimeric (partially humanized) murine monoclonal antibody glycoprotein that binds specifically to human tumor necrosis factor alpha (TNF-alpha) with an association constant of 1010 M-I. TNF-alpha is a naturally occurring cytokine that is involved in normal inflammatory and immune responses.
Elevated concentrations of TNF-alpha are known to exist in the joints of RA (Rheumatoid Arthritis) patients and the stools of Crohn's disease patients. In RA, Infliximab reduces the infiltration of inflammatory cells into inflamed areas of the joints and also reduces expression of molecules mediating cellular adhesion, chemo attraction and tissue degradation e.g. ICAM-I, IL-8, MCP-I, MMP-I & 3 etc. In Crohn's disease also, Infliximab reduces infiltration of inflammatory cells and decreases TNF-alpha production in inflamed areas of the intestine.
Infliximab, in combination with methotrexate, is indicated for the reduction in signs and symptoms of RA in patients who have had an inadequate response to methotrexate. Infliximab is also indicated for the reduction in signs and symptoms of Crohn's disease in patients with moderately to severely active Crohn's disease who have had an inadequate response to conventional therapy.
It is evident that Infliximab can be used to treat multiple health ailments and hence it is required to be produced on a commercial large scale set up. Challenges in the purification of proteins viz. monoclonal antibodies (MAbs) include reducing production cost, developing robust processes for both product purity and viral clearance, and integrating upstream and downstream processes. Purifying MAbs for therapeutic use requires highly selective and robust technologies to achieve the very high purity required for biopharmaceuticals. Processing such valuable product streams at large-scale while manufacturing to such high standards requires careful technology design and execution. This is because supernatant containing the target MAbs are most often obtained from the culture of live cells and are variable in both product content and composition. The lowest production cost is achieved not by increased process complexity but by the opposite: Operating faster and with the fewest and most efficient downstream steps reduces investments in validation and subsequent batch failures (Kelley B. Very large scale monoclonal antibody purification: the case for conventional unit operations. Biotechnology Programme. 2007; 23(5):995-1008). The present invention constitutes a pathbreaking improvement in purification and recovery of Recombinant Infliximab.
OBJECTIVES OF THE INVENTION
The main object of the present invention is to use novel fermentation and chromatographic procedures for rapid and efficient recovery of Recombinant Infliximab, a monoclonal antibody to human TNF-alpha, from cell culture supernatant. This invention is directed to a process for facilitating maximum product (recombinant Infliximab) recovery.
SUMMARY OF THE INVENTION
The present invention relates to the use of novel fermentation process for the over expression of Recombinant Infliximab, a monoclonal antibody to human TNF-alpha, protein in CHO cells.
The present invention also relates to the use of novel chromatographic procedures separately and jointly for the production of Recombinant Infliximab, a monoclonal antibody to human
TNF-alpha, protein, in biologically active form from fluids, especially mammalian host cell culture supernatants.
DETAILED DESCRIPTION OF THE DRAWINGS
Figure 1: Process chromatogram of affinity chromatography: Chromatographic profile of the first step of recovery of the target protein from the affinity chromatography. This is the purification chromatogram from a supernatant containing Infliximab as explained in example 1 (Supernatant derived from a harvested media containing Infliximab after clarification). The blue line represents the UV absorbance at 280nm. Majority of the other protein comes out in the flow through as can be observed in the figure.
Figure 2: Process chromatogram of anion chromatography: Chromatographic profile of the second step of recovery of the target protein from the anion exchange chromatography. This figure shows a chromatogram of purification from the eluent of the affinity chromatography as explained in the example 1 (Start material on to this column was the buffer exchanged neutralized elute containing the infliximab from the affinity column). It is clear from the figure that the majority of the load has come out in the flow through mode. A minor peak in the end may be some amount of the inflixmab has bound, as it is clear from the recovery that the majority of it has come out in follow-through.
Figure 3: Process chromatogram of cation chromatography: Chromatographic profile of the final step of recovery of the target protein from the cation exchange chromatography, This is a chromatogram from step 3 of a process according to the invention and it is more specifically a polishing step as explained in the example 1. The extra peak at the end is strongly bound protein that comes off with time.
Fig 4 : Electrophoretic pattern of Drug substance: showing comparable molecular weight with RMP where Lane No. 1 : Molecular weight Marker, Lane No. 2 : RMP and Lane No. 3 : Formulated Drug Substance. Electrophoretic profile reveals the purity and identity of the target protein with the Originator. The electrophoretic diagram depicts the stability of the drug substance and also the purity of the process close to homogeneity and comparability of the drug substance with that of the Originator.
Fig 5: Protein A HPLC profile of Drug substance showing comparable with RMP: This profile shows similar retention time in the protein A column. This is one of the orthogonal methods for estimation of target protein (recombinant inflximab). This figure is an analytical protein A column chromatogrm of the purified drug substance containing inflximab as it was compared with that of the originator. The extra peak seen at the beginning of the chromatograms is from the buffer as it can be observed in both the chromatogram (innovators as well as the inflximab purified product). This method not only gives the protein estimation but also the purity of the product with respect to the RMP {Reference Medicinal Product - Remicade®)
DETAILED DESCRIPTION OF THE INVENTION
The present invention relates to a process for the recovery of Recombinant Monoclonal antibody to TNF alpha from cell culture supernatant arising out of cell culture fluid by means of an innovative series of chromatographic steps. The present invention also relates to the recombinant Monoclonal antibody to TNF alpha purification procedure involving serial application different chromatographic techniques as mentioned previously. AU different steps, conditions and compositions are disclosed in the invention Culture supernatants are clarified before chromatographic treatment. First affinity chromatography is used for capture of recombinant Monoclonal antibody to TNF alpha. This process comprises selective binding of the desired compound to specific affinity resin and then elution with elution buffer. Resultant monoclonal antibody to TNF alpha containing eluent fractions are still enriched with biologically active material. So they are further subjected to processing by Ion exchange chromatographic step. Anion exchange and cation exchange chromatography are used for removal of process related impurities like host cell protein and host cell DNA.
In an advantageous embodiment of the present method, it is found that the multi-modal chromatography capture step followed by ion exchange chromatography steps yield a product that increases its step recovery from 94.46% in Step 1 to 98.65% in Step 3. [Table I].
The present invention will be more clearly understood from a consideration of the specific examples that follow.
Example 1:
The cell culture harvest is clarified by using a cellulose disposable filter. This filter is expressly customized with the effective filtration area being 650-1000 cm2 and an operating pressure of up to 30 psi. The filtrate is checked for turbidity and target protein content. Affinity chromatography is used in binding and elution mode with column of 32 mm diameter for capturing; with Tris buffer pH 7.2 - 7.6 as equilibration buffer. After the sample
is loaded on to the column, it is washed with equilibration buffer followed by 50 mM Tris-Cl. Then the second wash is performed with 50 mM Tris-Cl containing 100-400 mM NaCl pH 7-8 buffer solution. The protein of interest is eluted with citrate buffer pH 3.0-3.8 (Fig 1). The eluate is held for 45 - 60 min at acidic pH at room temperature for virus inactivation and later neutralized. The Protein A eluate fraction is taken for next purification step. Anion exchange chromatography in negative binding mode is then carried out at an operational flow rate of 140 cm/hr. The column has been equilibrated with Tris buffer pH 6.8 - 7.2. Protein of interest is collected in flow through. This step is used for the removal of process related impurities like leachate protein A, host cell DNA and host cell protein. (Fig 2). Thereafter, the flow through is filtered for virus removal using viral removal filter having an effective filtration area of 0.01 m2. The filtrate is then buffer exchanged using a 50 kDa TFF membrane. The buffer used for the diafiltration process is Tris buffer ph 6.8-7.2. Cation exchange chromatography is carried out with the diafiltered protein solution after equilibrating the column with Tris buffer pH 6.8-7.2. The protein of interest is eluted with elution buffer using NaCl salt gradient. This step is used for the removal of process related impurities like host cell DNA and host cell protein, which are there in traces (Fig 3). The eluate is buffer exchanged and concentrated using a 50 kDa TFF membrane at a Trans Membrane Pressure (TMP) of 5 - 10 psi. The buffer exchanged protein solution is filtered using 0.2μm filter. The drug substance was characterized as per the specifications. The Drug Substance (Active Pharmaceutical Ingredient) is formulated using formulation buffer pH 7.2, containing following constituents contains, 500 mg sucrose, 0.5 mg polysorbate 80, 2.2 mg monobasic sodium phosphate, monohydrate, and 6.1 mg dibasic sodium phosphate, dihydrate yielding a concentration of 10mg/ml of Infliximab. No preservatives are present. The efficiency of the process is depicted in terms of the recovery per say of the target protein. (Table 1).
Table 1: Depicts the percentage of recovery of the target protein in purification process.
Example 2:
The formulated material is characterized as per the specifications set by product development specification so as to meet the physico-chemical parameter comparable with that of the
originator. A 10% SDS PAGE under reducing condition is studied for the sample derived from the PAGE showed a clear corresponding band with RMP (Reference Medicinal Product -Remicade®) reflecting the fact that there is a comparable purity and integrity (Fig 4). Protein A HPLC profile carried out during this step showed in test molecule, which was very much similar to the RMP. This technique is used as one of the orthogonal method for estimation of target protein (Fig 5).
Claims
1. A process for the recovery and purification of recombinant Monoclonal antibody to TNF alpha comprising steps of: contacting culture supernatant(s) with resin(s) for selective adsorption of compound(s); eluting the adsorbed compound and subjecting the enriched product to series of separation techniques.
2. The process as claimed in claim 1, where elution of adsorbed compound is performed by affinity chromatography.
3. The process as claimed in claim 1, where monoclonal antibody to TNF alpha containing eluent fractions are purified using ion exchange chromatography.
4. The process as claimed in claim 1, wherein supernatant is obtained from cell fermentation.
5. The process as claimed in claim 1, wherein said supernatant is mammalian host cell culture supernatant.
6. The process as claimed in claim 1, wherein said supernatant is cell culture-derived fluid.
7. The process as claimed in claim 1, wherein said supernatant is mammalian cell culture derived fluid
8. The process as claimed in claim 1, wherein the process buffer 20-100 mM Tris buffer pH 6.4-7.2 with 100-400 mM NaCl is used for the recovery of target protein from culture supernatant.
9. The process as claimed in claim 1, wherein the process buffer is particularly 40-60 mM Tris buffer pH 6.6-7.0 with 200-30OmM NaCl is used for the recovery of target protein from culture supernatant.
10. The process as claimed in claim 1, wherein said culture supernatant(s) are concentrated and clarified before contacting resins.
11. The process as claimed in claim 1, where eluent is held for 45 - 60 min at acidic pH at room temperature for virus inactivation.
12. The process as claimed in claim 1, where recombinant Monoclonal antibody to TNF alpha recovered and purified is Infliximab.
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2015198320A1 (en) | 2014-06-24 | 2015-12-30 | Insight Biopharmaceuticals Ltd. | Methods of purifying antibodies |
EP2691411B1 (en) | 2011-03-29 | 2020-02-26 | GlaxoSmithKline LLC | Buffer system for protein purification |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5698195A (en) * | 1991-03-18 | 1997-12-16 | New York University Medical Center | Methods of treating rheumatoid arthritis using chimeric anti-TNF antibodies |
WO2004050683A2 (en) * | 2002-12-02 | 2004-06-17 | Abgenix, Inc. | Antibodies directed to tumor necrosis factor and uses thereof |
US20080255027A1 (en) * | 2006-12-21 | 2008-10-16 | Wilson Moya | Purification of proteins |
-
2010
- 2010-08-02 WO PCT/IB2010/001896 patent/WO2011015919A1/en active Application Filing
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5698195A (en) * | 1991-03-18 | 1997-12-16 | New York University Medical Center | Methods of treating rheumatoid arthritis using chimeric anti-TNF antibodies |
WO2004050683A2 (en) * | 2002-12-02 | 2004-06-17 | Abgenix, Inc. | Antibodies directed to tumor necrosis factor and uses thereof |
US20080255027A1 (en) * | 2006-12-21 | 2008-10-16 | Wilson Moya | Purification of proteins |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2691411B1 (en) | 2011-03-29 | 2020-02-26 | GlaxoSmithKline LLC | Buffer system for protein purification |
WO2015198320A1 (en) | 2014-06-24 | 2015-12-30 | Insight Biopharmaceuticals Ltd. | Methods of purifying antibodies |
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