Modulation of charge variants in a monoclonal antibody composition
Inventors
YONAN, Chris • CAROSELLI, Christine • DENDAMRONGVIT, Wiphusanee • Gangloff, Scott
Assignees
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Publication Number
US-10696735-B2
Publication Date
2020-06-30
Expiration Date
Abstract
Combinations of different chromatography modalities with particularly refined conditions significantly reduce acid charge variants in a preparation of monoclonal antibodies. The process for reducing acid charge variants utilizes a combination of anion exchange and hydrophobic interaction chromatography, followed by cation exchange chromatography polishing, whereby the levels of acidic or basic charge species of the monoclonal antibodies may be modulated to a desired level.
Core Innovation
The invention provides a process for removing acid charge variants from a monoclonal antibody. A mammalian cell-expressed monoclonal antibody preparation is loaded onto a Protein A support and eluted to produce a first eluate comprising the monoclonal antibody. The first eluate is then directed to an anion exchange and hydrophobic interaction (AEX/HIC) chromatography support such that it flows through to form a flow-through pool comprising the monoclonal antibody.
The flow-through pool is loaded onto a cation exchange (CEX) chromatography support having an antibody binding capacity of from about 25 g/L to about 65 g/L. The process includes a first wash step, a second wash step, and a third wash step with specified pH and conductivity targets, and the second wash step includes determining when UV A280 absorbance units decrease from about 7% to about 14% from the peak absorbance units measured at UV A280.
The monoclonal antibody is eluted from the CEX chromatography support using an elution buffer having a pH from about 6.0 to about 6.4 and a conductivity target of from about 10 mS/cm to about 14 mS/cm. The elution produces a second eluate comprising the monoclonal antibody and from about 10% to about 20% by weight of acid charge variants of the monoclonal antibody.
Claims Coverage
The partial content provides one independent claim directed to removing acid charge variants from a monoclonal antibody using a sequential Protein A, AEX/HIC flow-through, and CEX wash/elution scheme with UV A280-driven wash control and defined target levels for acid charge variants. Three inventive features are identified from the claim excerpts.
Sequential Protein A capture followed by AEX/HIC flow-through pool formation
Loading a mammalian cell-expressed monoclonal antibody preparation onto a Protein A support, eluting to produce a first eluate, and loading the first eluate onto an anion exchange and hydrophobic interaction (AEX/HIC) chromatography support such that the first eluate flows through to produce a flow-through pool comprising the monoclonal antibody.
CEX washing with UV A280-based determination of wash termination
Loading the flow-through pool onto a cation exchange (CEX) chromatography support having an antibody binding capacity of from about 25 g/L to about 65 g/L and performing first, second, and third wash steps with specified pH and conductivity targets, including a second wash step in which UV A280 absorbance units decrease from about 7% to about 14% from peak absorbance units.
CEX elution to yield a second eluate with defined acid charge variant level
Eluting the monoclonal antibody from the CEX chromatography support with an elution buffer having a pH of from about 6.0 to about 6.4 and a conductivity target of from about 10 mS/cm to about 14 mS/cm to produce a second eluate comprising the monoclonal antibody and from about 10% to about 20% by weight of acid charge variants.
Overall, the claim coverage centers on a sequential workflow of Protein A capture, AEX/HIC flow-through processing, and CEX wash/elution with UV A280-based control and defined pH and conductivity conditions to produce a second eluate having about 10% to about 20% by weight acid charge variants.
Stated Advantages
Produces a second eluate comprising the monoclonal antibody and from about 10% to about 20% by weight of acid charge variants.
Documented Applications
Not explicitly described in patent.
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