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.
Claims40
A method for producing at least two different antibodies in a single host cell, said method comprising,
- (a) providing a host cell comprising
- (i) A first nucleic acid molecule encoding a 1st antibody heavy chain with a CH3 domain comprising the amino acid substitution T366K,
- (ii) A second nucleic acid molecule encoding a 2nd antibody heavy chain with a CH3 domain comprising the amino acid substitution L351D,
- (iii) A third nucleic acid molecule encoding a 3rd antibody heavy chain, and
- (iv) A fourth nucleic acid molecule encoding a 4th antibody heavy chain,
- (b) culturing said host cell and allowing for expression of said antibody heavy chains, wherein the CH3 domain of said 1st antibody heavy chain preferentially pairs with the CH3 domain of said 2nd antibody heavy chain, and the CH3 domain of said 3rd antibody heavy chain preferentially pairs with the CH3 domain of said 4th antibody heavy chain to produce antibodies containing said 1st and 2nd antibody heavy chains and said 3rd and 4th antibody heavy chains;
- and
- (c) harvesting said at least two different antibodies from the culture.
A recombinant host cell comprising nucleic acids encoding at least a first, a second, a third and a fourth 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 expression of said first, second, third and fourth antibody heavy chains results in preferential pairing of said first antibody heavy chain with said second antibody heavy chain, and preferential pairing of said third antibody heavy chain with said fourth antibody heavy chain.
A method for making a host cell which produces at least two different antibodies, the method comprising the step of introducing into said host cell nucleic acids encoding at least a first, a second, a third and a fourth antibody heavy chain, wherein the CH3 domain of said first antibody heavy chain comprises the amino acid substitution T366K and CH3 domain of said second antibody heavy chain comprises the amino acid substitution L351D, wherein said nucleic acids are introduced consecutively or concomitantly, and wherein expression of said first, second, third and fourth antibody heavy chains in said host cell results in preferential pairing of said first antibody heavy chain with said second antibody heavy chain, and preferential pairing of said third antibody heavy chain with said fourth antibody heavy chain.
Dependencies are inferred from the claim text.