Methods and means for the production of Ig-like molecules
AssigneeMerus N.V.
InventorsCornelis A. 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.
Claims37
A method for producing a heterodimeric antibody from a single cell, wherein said antibody comprises two heavy chains with CH3 domains that are capable of forming an interface, said method comprising:
- a. providing a host cell comprising (i) a first nucleic acid molecule encoding a 1st antibody heavy chain comprising the amino acid substitution T366K in the CH3 domain, and (ii) a second nucleic acid molecule encoding a 2nd antibody heavy chain comprising the amino acid substitution L351D in the CH3 domain
- b. culturing said host cell and allowing for expression of said two nucleic acid molecules to produce said 1st antibody heavy chain and said 2nd antibody heavy chain, wherein the T366K substituted amino acid in the CH3 domain of said 1st antibody heavy chain interacts with the L351D substituted amino acid in the CH3 domain of said 2nd antibody heavy chain in the interface between the CH3 domains of said 1st and 2nd antibody heavy chain to produce a heterodimeric antibody;
- and
- c. harvesting said heterodimeric antibody from the culture.
A heterodimeric antibody comprising two CH3 domains, wherein one of said two CH3 domains comprises the amino acid substitutions L351D and L368E and wherein the other of said two CH3 domains comprises the amino acid substitutions T366K and L351K.
A recombinant host cell comprising nucleic acid sequences encoding a 1st and 2nd antibody heavy chain, wherein the CH3 domain of said 1st antibody heavy chain comprises the amino acid substitution T366K, and wherein the CH3 domain of said 2nd antibody heavy chain comprises the amino acid substitution L351D.
A method for making a host cell for production of a heterodimeric antibody, the method comprising the step of introducing into said host cell nucleic acid molecules encoding at least a first and a second antibody heavy chain, wherein the CH3 domain of said first antibody heavy chain comprises the amino acid substitution T366K, and the CH3 domain of said second antibody heavy chain comprises the amino acid substitution L351D, and wherein said nucleic acid molecules are introduced consecutively or concomitantly.
A heterodimeric antibody comprising two CH3 domains, wherein one of said two CH3 domains comprises the amino acid substitution L351D and wherein the other of said two CH3 domains comprises the amino acid substitution T366K.
Dependencies are inferred from the claim text.