Modulation of complement-dependent cytotoxicity through modifications of the c-terminus of antibody heavy chains
AssigneeGenmab A/S
InventorsPaul Parren, Patrick Van Berkel, Ewald T.J. Van Den Bremer
Life of this patent
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Abstract
Antibody variants having decreased or increased ability to mediate CDC due to modifications at the C-terminus of their heavy chains are described. Methods of generating such antibodies, as well as nucleotide constructs and host cells suitable for the production of said antibodies are also described.
Claims41
A method for producing an antibody in a host cell, wherein said antibody comprises at least a heavy chain, said method comprising the following steps:
- a) providing a nucleotide construct encoding said heavy chain, wherein said construct does not encode a lysine residue at the C-terminus of said heavy chain,
- b) expressing said nucleotide construct in a host cell, with the proviso that said host cell is not a CHO cell or a moss cell, and
- c) recovering said antibody from a cell culture of said host cell.
3-8. (canceled)
(canceled)
A host cell which is not capable of efficiently removing C-terminal lysine residues from antibody heavy chains, said host cell comprising a nucleotide construct encoding a heavy chain lacking a C-terminal lysine.
A method for increasing the ability of an antibody to mediate complement-dependent cytotoxicity, said method comprising the step of removing a C-terminal lysine residue from the heavy-chains of an antibody.
17-18. (canceled)
An isolated antibody variant comprising a heavy chain having at least one charged amino acid residue in the C-terminal region, wherein said charged amino acid residue is not susceptible to proteolytic removal.
24-25. (canceled)
27-31. (canceled)
33-39. (canceled)
A nucleotide construct encoding a heavy chain having a charged amino acid residue in the C-terminal region, wherein said charged amino acid residue is not susceptible to proteolytic removal.
A host cell which has been genetically modified to eliminate the activity of carboxypeptidases capable of removing C-terminal lysines from heavy chains.
An antibody mixture comprising a subpopulation of antibody variant molecules comprising heavy chains having at least one positively-charged amino acid residue in the C-terminal region and a subpopulation of antibody variants molecules comprising heavy chains having at least one negatively-charged amino acid residue C-terminal region, wherein said positively-charged and negatively-charged amino acid residues are not susceptible to proteolytic removal.
49-50. (canceled)
An isolated antibody comprising a heavy chain having a variant C-terminal region, comprising one or more amino acid modifications that favour intermolecular C-terminal interactions between antibody molecules.
(canceled)
56-57. (canceled)
Dependencies are inferred from the claim text.