Kappa light chain-binding convection matrix
AssigneeCytiva BioProcess R&D AB
InventorsRonnie Palmgren, Keyhan Esfandiarfard, Tania Ahmad, Tomas Björkman, Lisa Marx
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Abstract
The present invention relates to isolation and separation of kappa light chain-containing proteins. More specifically it relates to a separation matrix comprising kappa light chain-binding ligands covalently coupled to a porous support, wherein said kappa light chain-binding ligands comprise, consists essentially of, or consists of multimers of alkali-stabilized Finegoldia magna (formerly Peptostreptococcus Magnus) Protein L domains; and said porous support is a convection-based chromatography matrix.
Claims20
A separation matrix comprising kappa light chain-binding ligands covalently coupled to a porous support, wherein
- said kappa light chain-binding ligands comprise, consists essentially of, or consists of multimers of alkali-stabilized Finegoldia magna (formerly Peptostreptococcus Magnus) Protein L domains;
- and
- said porous support is a convection-based chromatography matrix.
A method of isolating a kappa light chain-containing protein comprising the steps of:
- a) contacting a liquid sample comprising a kappa light chain-containing protein with a separation matrix;
- b) washing said separation matrix with one or a combination of several washing liquids;
- c) eluting the kappa light chain-containing protein from the separation matrix with an elution liquid;
- and
- d) cleaning the separation matrix with a cleaning liquid;
- wherein the separation matrix has a dynamic binding capacity (DBC) of a kappa light chain-comprising antibody, such as Trastuzumab, of 25 g/mL at 10% breakthrough when running at a flow of 10 mL/min in a 0.4 mL HiTrap™ device.
A method for separation of isolating a kappa light chain-binding protein from lambda light chain-containing proteins, comprising the steps of
- a) contacting a liquid sample comprising a kappa light chain-containing protein with a separation matrix;
- b) washing said separation matrix with one or a combination of several washing liquids;
- c) eluting the kappa light chain-containing protein from the separation matrix with an elution liquid;
- and
- d) cleaning the separation matrix with a cleaning liquid;
- wherein the separation matrix has a dynamic binding capacity (DBC) of a kappa light chain-comprising antibody, such as Trastuzumab, of 25 g/mL at 10% breakthrough when running at a flow of 10 mL/min in a 0.4 mL HiTrap™ device.
A method for separation of bispecific antibodies from mono-specific antibodies, comprising the steps of:
- a) contacting a liquid sample comprising a bispecific antibody with a separation matrix;
- b) washing said separation matrix with one or a combination of several washing liquids;
- c) eluting the bispecific antibody from the separation matrix with an elution liquid;
- and
- d) cleaning the separation matrix with a cleaning liquid;
- wherein the separation matrix has a dynamic binding capacity (DBC) of a kappa light chain-comprising antibody, such as Trastuzumab, of 25 g/mL at 10% breakthrough when running at a flow of 10 mL/min in a 0.4 mL HiTrap™ device.
(canceled)
Dependencies are inferred from the claim text.