Methods and means for the production of ig-like molecules
AssigneeMerus N.V.
InventorsCornelis Adriaan DE KRUIF, Linda Johanna Aleida HENDRIKS, Ton LOGTENBERG
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Abstract
The invention provides means and methods for producing one or more Ig-like molecules in a single host cell. Novel CH3 mutations enabling the production of monospecific and/or bispecific Ig-like molecules of interest are also provided.
Claims22
-28. (canceled)
A recombinant host cell comprising nucleic acid sequences encoding at least a first antibody polypeptide chain and a second antibody polypeptide chain, each comprising CH1, CH2 and CH3 domains,
- wherein the CH3 domain of said first antibody polypeptide chain comprises at least one substitution of a neutral amino acid residue by a positively charged amino acid residue, wherein the CH3 domain of said second antibody polypeptide chain comprises at least one substitution of a neutral amino acid residue by a negatively charged amino acid residue, wherein the at least one positively charged amino acid residue substituted in the CH3 domain of said first antibody polypeptide chain interacts with the at least one negatively charged amino acid residue substituted in the CH3 domain of said second antibody polypeptide chain at the interface between said first and second antibody polypeptide chains, and
- wherein the interaction between the at least one positively charged amino acid residue substituted in the CH3 domain of said first antibody polypeptide chain and the at least one negatively charged amino acid residue substituted in the CH3 domain of said second antibody polypeptide chain produces a heterodimeric antibody.
A recombinant host cell comprising nucleic acid sequences encoding at least a first antibody polypeptide chain and a second antibody polypeptide chain, each comprising CH1, CH2 and CH3 domains,
- wherein the CH3 domain of said first antibody polypeptide chain comprises a substitution of the amino acid residue at position 364 according to the EU numbering system by a positively charged amino acid residue,
- wherein the CH3 domain of said second antibody polypeptide chain comprises at least one substitution of a neutral amino acid residue by a negatively charged amino acid residue, wherein the positively charged amino acid residue substituted at the position 364 in the CH3 domain of said first antibody polypeptide chain interacts with the at least one negatively charged amino acid residue substituted in the CH3 domain of said second antibody polypeptide chain at the interface between said first and said second antibody polypeptide chains, and
- wherein the interaction between the at least one positively charged amino acid residue substituted in the CH3 domain of said first antibody polypeptide chain and the at least one negatively charged amino acid residue substituted in the CH3 domain of said second antibody polypeptide chain produces a heterodimeric antibody.
A method for making a host cell comprising introducing into said host cell nucleic acid molecules encoding a first antibody polypeptide chain and a second antibody polypeptide chain, each comprising CH1, CH2 and CH3 domains,
- wherein the CH3 domain of said first antibody polypeptide chain comprises at least one substitution of a neutral amino acid residue by a positively charged amino acid residue, wherein the CH3 domain of said second antibody polypeptide chain comprises at least one substitution of a neutral amino acid residue by a negatively charged amino acid residue, wherein the at least one positively charged amino acid residue substituted in the CH3 domain of said first antibody polypeptide chain interacts with the at least one negatively charged amino acid residue substituted in the CH3 domain of said second antibody polypeptide chain at the interface between said first and said second antibody polypeptide chains,
- wherein the interaction between the at least one positively charged amino acid residue substituted in the CH3 domain of said first antibody polypeptide chain and the at least one negatively charged amino acid residue substituted in the CH3 domain of said second antibody polypeptide chain produces a heterodimeric antibody, and
- wherein said nucleic acid sequences are introduced consecutively or concomitantly.
A method for making a host cell comprising introducing into said host cell nucleic acid molecules encoding a first antibody polypeptide chain and a second antibody polypeptide chain, each comprising CH1, CH2 and CH3 domains,
- wherein the CH3 domain of said first antibody polypeptide chain comprises a substitution of an amino acid residue at position 364 according to the EU numbering system by a positively charged amino acid residue,
- wherein the CH3 domain of said second antibody polypeptide chain comprises at least one substitution of a neutral amino acid residue by a negatively charged amino acid residue, wherein the positively charged amino acid residue substituted at the position 364 in the CH3 domain of said first antibody polypeptide chain interacts with the at least one negatively charged amino acid residue substituted in the CH3 domain of said second antibody polypeptide chain at the interface between said first and said second antibody polypeptide chains, and
- wherein the interaction between the at least one positively charged amino acid residue substituted in the CH3 domain of said first antibody polypeptide chain and the at least one negatively charged amino acid residue substituted in the CH3 domain of said second antibody polypeptide chain produces a heterodimeric antibody.
Dependencies are inferred from the claim text.