Patent ID: 9163249
Filing Date: 2015-10-20
Classification: A61K,A61P,C07K,C12N

Abstract:
1. A method to produce a mammalian final cell line producing at least one therapeutic protein comprising the steps of: (a) obtaining a first polynucleotide sequence having a codon adaptation index less than 0.9and that encodes said at least one therapeutic protein; (b) altering the first polynucleotide sequence to obtain a second polynucleotide sequence, wherein the codon adaptation index of the second polynucleotide sequence is equal to or greater than 0.9 and the first polynucleotide and second polynucleotide encode the same therapeutic protein; (c) transforming at least one first cell with the first polynucleotide sequence of step (a) and a third polynucleotide sequence that encodes a selection marker which is capable of providing amplification of the first polynucleotide sequence within said first cell; and transforming at least one second cell with the second polynucleotide sequence of step (b) and said third polynucleotide sequence that encodes a selection marker which is capable of providing amplification of the second polynucleotide sequence within said second cell; (d) growing said at least one first cell of step (c) to create a first cell line comprising a plurality of cells, in medium that contains a selection and amplification agent; growing said at least one second cell of step (c) to create a second cell line comprising a plurality of cells, in medium that contains a selection and amplification agent at a concentration of 50% or less than is necessary to reach an equivalent plateau of production of said protein produced in said first cell line transformed with the first polynucleotide; and (e) selecting said final cell line from said second cell line once the plateau of production of the protein encoded by the second polynucleotide in said second cell line is reached.