Increasing The Production Of Recombinant Antibodies In Mammalian Cells By Site-Directed Mutagenesis
AssigneeMedImmune, LLC
InventorsWilliam DALL'ACQUA, Herren WU, Melissa DAMSCHRODER
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Abstract
The present invention relates to a reliable, reproducible method for improving the producibility of an antibody. More specifically, this invention provides a method for modifying the heavy chain of an antibody to improve its producibility in eukaryotic cells. Additionally, the method of the invention may improve both antibody producibility and one or more antigen binding characteristics. The invention further provides modified antibodies which are better produced and which have either no change in their antigen binding characteristics or exhibit improved antigen binding characteristics.
Claims20
A method of increasing the production of an antibody from a eukaryotic host cell, wherein said method comprises the steps of:
- (a) introducing one or more mutations into a nucleotide sequence encoding the antibody heavy chain, wherein said one or more mutations result in the substitution of one or more of the amino acid residues selected from the group consisting of:
- positions 40H, 60H, and 61H, utilizing the numbering system set forth in Kabat, with alanine, alanine, and aspartic acid, respectively;
- (b) introducing into the eukaryotic host cell the nucleotide sequence encoding the heavy chain of the antibody and a nucleotide sequence encoding the antibody light chain;
- and
- (c) cultivating the eukaryotic host cell under conditions wherein the antibody comprising the substitution is expressed by said eukaryotic host cell, wherein said antibody is a full length antibody;
- and wherein production of the antibody comprising the substitution is increased by at least 1.3 fold and the binding specificity is unchanged compared to the antibody without the substitution.
An antibody comprising a substitution of one or more amino acid residues selected from the group consisting of:
- positions 40H, 60H, and 61H, utilizing the numbering system set forth in Kabat, with alanine, alanine, and aspartic acid, respectively;
- wherein said antibody is a full length antibody;
- and wherein production of the antibody comprising the substitution in a eukaryotic host cell is increased by at least 1.3 fold and the binding specificity is unchanged compared to the antibody without the substitution.
Dependencies are inferred from the claim text.