Cell lines
AssigneeMedImmune, LLC
InventorsKenneth H. Grabstein, Michael Van Brunt, Marcello Marelli, William Brady, Jeffrey C. Johnson
Life of this patent
Estimating the term…
Abstract
There is provided inter alia a process for stabilizing a eukaryotic cell line which expresses PylRS and tRNAPyl and which is suitable for incorporation of a gene encoding a target protein containing one or more non-natural amino acids encoded by a nonsense codon which comprises culturing said cell line under conditions in which the adverse effect of tRNAPyl expression on cell viability and/or cell growth is reduced or eliminated.
Claims6
A process for stabilizing a eukaryotic cell line which expresses PylRS and tRNAPyl, wherein the cell line is capable of incorporating a gene encoding a target protein and wherein said target protein comprises one or more non-natural amino acids encoded by an amber codon, wherein said process comprises culturing said cell line in the presence of a decoy amino acid of:
- (a) formula VIIA:
- wherein K is CO or SO2;
- Q=H, C1-6alkyl, aryl, heteroaryl, -OC1-6alkyl, -OCH2aryl, -OCH2heteroaryl, C2-6alkenyl or -OC2-6alkenyl;
- or
- (b) formula VIIB:
- wherein G=H;
- a=4 or 5;
- and
- R=C1-6alkyl, C2-6alkenyl, CH2aryl, C2-6alkynyl, C1-6haloalkyl or C1-6azidoalkyl,
- wherein the viability of said eukaryotic cell line is enhanced in comparison to the viability of said eukaryotic cell line cultured in the absence of said decoy amino acid.
Dependencies are inferred from the claim text.