Method for purifying antibodies
Inventors
Meh, David • Atolagbe, Timothy • Farquharson, G. Mark • Shaban, Samir • Koleck, Mary • Mitra, George
Assignees
United Therapeutics Corp • US Department of Health and Human Services
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Publication Number
US-10906935-B2
Publication Date
2021-02-02
Expiration Date
2035-07-27
Abstract
Disclosed here includes a method for purifying a biologic composition, comprising diafiltering the biologic composition into a composition comprising phosphate buffered saline (PBS) to obtain a purified composition. The method disclosed here can be particularly useful for removing one or more impurities from the biologic composition, such as bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane (Bis-tris).
Core Innovation
The invention provides a method for purifying a biologic composition by diafiltering the biologic composition into a composition comprising phosphate buffered saline (PBS) to obtain a purified composition. This method specifically addresses the removal of impurities such as bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane (Bis-tris) from biologic compositions including monoclonal antibodies like ch14.18.
The problem addressed is that known purification processes for monoclonal antibodies need effective removal of unwanted impurities, especially for therapeutic uses where safety and effectiveness are critical. Bis-tris, used as a buffer in chromatography steps such as with Capto™ adhere resin, can remain as a contaminant after purification. Previous diafiltration directly into formulation buffers failed to remove Bis-tris effectively.
The solution is the use of diafiltration with PBS, which was shown to efficiently remove Bis-tris from the biologic composition, including ch14.18 antibody preparations. This step can be integrated after chromatographic purification steps, particularly after elution with Bis-tris buffer from an anion exchange column, improving purification by substantially eliminating Bis-tris impurities prior to final formulation.
Claims Coverage
The patent contains two independent claims that cover the main inventive methods disclosed.
Diafiltration of biologic composition with PBS for purification
A method comprising diafiltering a biologic composition containing Bis-tris impurity and ch14.18 monoclonal antibody using phosphate buffered saline (PBS) to obtain a purified composition with reduced Bis-tris.
Comprehensive monoclonal antibody purification process including PBS diafiltration
A method for purifying a monoclonal antibody involving sequential steps: affinity chromatography, pH lowering, solvent-detergent washing, cation exchange chromatography, nano-filtration, anion exchange chromatography yielding a composition with Bis-tris, and diafiltration into PBS to obtain a composition substantially free from Bis-tris.
The patent claims center on the purification of ch14.18 monoclonal antibody compositions using diafiltration with PBS to remove Bis-tris impurities, either as a standalone purification step or integrated into a multistep antibody purification process, highlighting the effective removal of Bis-tris and improvement of composition purity.
Stated Advantages
Diafiltration with PBS effectively removes Bis-tris impurities from biologic compositions containing monoclonal antibodies.
The method improves safety and effectiveness of therapeutic monoclonal antibodies by removing residual buffers such as Bis-tris after chromatographic purification steps.
PBS diafiltration allows for improved removal of impurities that are not efficiently removed by direct diafiltration into other formulation buffers like histidine-buffered saline (HBS).
Documented Applications
Purification of monoclonal antibodies, particularly ch14.18, for therapeutic use.
Removal of Bis-tris impurity from biologic compositions after elution from anion exchange chromatography columns using Bis-tris buffer.
Integration into multistep antibody purification processes involving affinity chromatography, pH inactivation, solvent-detergent viral inactivation, ion exchange chromatography, nano-filtration, and final diafiltration into PBS and optionally histidine buffer formulations.
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