Methods and means for the production of ig-like molecules

A method for producing a heterodimeric antibody from a single cell, wherein said antibody comprises a first antibody polypeptide chain and a second antibody polypeptide chain, each comprising CH1, CH2 and CH3 domains, wherein the CH3 domains are capable of forming an interface, said method comprising:

• a. providing a host cell comprising
• (i) a first nucleic acid molecule encoding said first antibody polypeptide chain comprising at least one substitution of a neutral amino acid residue in the CH3 domain by a positively charged amino acid residue, and
• (ii) a second nucleic acid molecule encoding said second antibody polypeptide chain comprising at least one substitution of a neutral amino acid residue in the CH3 domain by a negatively charged amino acid residue;
• b. culturing said host cell and allowing for expression of said two nucleic acid molecules to produce said first antibody polypeptide chain and said second antibody polypeptide chain,
• 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 in the interface between said first and second antibody heavy 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;
• and
• c. harvesting said heterodimeric antibody from the culture.

A method for producing a heterodimeric antibody from a single cell, wherein said antibody comprises a first antibody polypeptide chain and a second antibody polypeptide chain each comprising CH1, CH2 and CH3 domains, wherein the CH3 domains are capable of forming an interface, said method comprising:

• a. providing a host cell comprising
• (i) a first nucleic acid molecule encoding said first antibody polypeptide chain comprising a substitution of an amino acid residue at position 364 in the CH3 domain according to the EU numbering system by a positively charged amino acid residue, and
• (ii) a second nucleic acid molecule encoding said second antibody polypeptide chain comprising at least one substitution of a neutral amino acid residue in the CH3 domain by a negatively charged amino acid residue;
• b. culturing said host cell and allowing for expression of said two nucleic acid molecules to produce said first antibody polypeptide chain and said second antibody polypeptide chain,
• wherein the positively charged amino acid residue substituted at 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 in the interface between said first and second antibody heavy chains, and
• wherein the interaction between the positively charged amino acid residue substituted at position 364 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
• c. harvesting said heterodimeric antibody from the culture.