Methods of producing anti-C5 antibodies
Inventors
Malanson, Hunter F. • ZINGARO, Kyle A. • GIRI, Anjil • WEAVER, Justin • Friedman, Abraham • Hunter, Jeffrey William • SIVANANDAM, Saranya • ZUGATES, Jeffrey • Godawat, Rahul
Assignees
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Publication Number
US-12312394-B2
Publication Date
2025-05-27
Expiration Date
Abstract
The present application relates to a method of producing an anti-C5 antibody (ravulizumab), wherein the method comprises: —culturing mammalian cells comprising a nucleic acid encoding the anti-C5 antibody in a cell culture production medium—Performing two or more steps selected from the group consisting of: a recovery step; purification by Protein A affinity chromatography, a low pH viral inactivation step; Purification by cation exchange chromatography; Purification by anion exchange chromatography; a virus reduction filtration step; and a concentration and diafiltration step.
Core Innovation
The invention relates to a sequential method of producing an anti-C5 antibody using mammalian cells comprising a nucleic acid encoding the anti-C5 antibody, where the anti-C5 antibody is produced in a cell culture production medium. The method includes a recovery step by filtering the cell culture production medium through a depth filter, followed by purification by Protein A affinity chromatography.
The method further includes a low pH viral inactivation step, followed by purification by cation exchange chromatography and purification by anion exchange chromatography. After chromatography purification, a virus reduction filtration step is performed, followed by a concentration and diafiltration step using a polysorbate 80 buffer.
The anti-C5 antibody is defined by specified CDR1, CDR2 and CDR3 heavy chain sequences set forth in SEQ ID NOs: 19, 18 and 3, respectively, and specified CDR1, CDR2 and CDR3 light chain sequences set forth in SEQ ID NOs: 4, 5 and 6, respectively. Additional described embodiments provide an end-to-end ravulizumab manufacturing sequence and associated formulation compositions and release specifications.
Claims Coverage
The independent claim covers a sequential antibody production process with eight inventive features, and defines the antibody by specific heavy-chain and light-chain CDR sequences using SEQ ID NOs.
Sequential production of an anti-C5 antibody with defined processing order
A method of producing an anti-C5 antibody in which culturing mammalian cells, recovery by depth filtration, Protein A affinity chromatography, low pH viral inactivation, cation exchange chromatography, anion exchange chromatography, virus reduction filtration, and concentration and diafiltration are performed sequentially in order.
Mammalian cell culturing in cell culture production medium
Culturing mammalian cells comprising a nucleic acid encoding the anti-C5 antibody in a cell culture production medium such that the anti-C5 antibody is produced in said cell culture production medium.
Protein A affinity purification after depth filtration
A recovery step comprising filtering the cell culture production medium through a depth filter, followed by purification by Protein A affinity chromatography.
Low pH viral inactivation integrated into the purification train
A low pH viral inactivation step performed after Protein A affinity chromatography and before the cation exchange and anion exchange chromatography steps.
Sequential cation and anion exchange chromatography
Purification by cation exchange chromatography followed by purification by anion exchange chromatography.
Virus reduction filtration after chromatography purification
A virus reduction filtration step performed after the purification by cation exchange chromatography and anion exchange chromatography steps.
Concentration and diafiltration using a polysorbate 80 buffer
A concentration and diafiltration step in which the concentration and diafiltration step involves using a polysorbate 80 buffer.
Anti-C5 antibody defined by specified CDR heavy and light chain sequences
The anti-C5 antibody comprises CDR1, CDR2 and CDR3 heavy chain sequences as set forth in SEQ ID NOs: 19, 18 and 3, respectively, and CDR1, CDR2 and CDR3 light chain sequences as set forth in SEQ ID NOs: 4, 5 and 6, respectively.
Overall, the claim coverage centers on the sequential combination of culturing and recovery, Protein A affinity purification, low pH viral inactivation, cation exchange then anion exchange chromatography, virus reduction filtration, and concentration/diafiltration using a polysorbate 80 buffer, together with an anti-C5 antibody defined by specific CDR sequences using SEQ ID NOs.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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