Codon engineering for improved antibody expression in mammalian cells.
While well established in bacterial hosts, the effect of coding sequence variation on protein expression in mammalian systems is poorly characterized outside of viral proteins or proteins from distant phylogenetic families. The potential impact is substantial given the extensive use of mammalian expression systems in research and manufacturing of protein biotherapeutics. We are studying the effect of codon engineering on expression of recombinant antibodies with an emphasis on developing manufacturing cell lines. CNTO 888, a human mAb specific for human MCP-1, was obtained by antibody phage display in collaboration with MorphoSys AG. The isolated DNA sequence of the antibody was biased towards bacterial codons, reflecting the engineering of the Fab library for phage display expression in Escherichia coli. We compared the expression of CNTO 888 containing the parental V-region sequences with two engineered coding variants. In the native codon exchanged (NCE) variant, the V-region codons were replaced with those used in naturally derived human antibody genes. In the human codon optimized (HCO) variant the V-region codons were those used at the highest frequency based on a human codon usage table. The antibody expression levels from stable transfections in mammalian host cells were measured. The HCO codon variant of CNTO 888 yielded the highest expressing cell lines and the highest average expression for the screened populations. This had a significant positive effect on the process to generate a CNTO 888 production cell line and indicates the potential to improve antibody expression in mammalian expression systems by codon engineering.
Carton
Jill M
JM
Department of Molecular Discovery Technologies, Centocor Inc., 145 King of Prussia Road, Radnor, PA 19087, USA. jcarton2@cntus.jnj.com
Sauerwald
Tina
T
Hawley-Nelson
Pam
P
Morse
Barry
B
Peffer
Nancy
N
Beck
Heena
H
Lu
Jin
J
Cotty
Adam
A
Amegadzie
Bernard
B
Sweet
Ray
R
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D002460:Cell Line; D003062:Codon; D004274:DNA, Recombinant; D005818:Genetic Engineering; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids
D000911:Antibodies, Monoclonal; D003062:Codon; D004274:DNA, Recombinant