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 exp...

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Bibliographic Details
Published in:Protein expression and purification 2007-10, Vol.55 (2), p.279-286
Main Authors: Carton, Jill M., Sauerwald, Tina, Hawley-Nelson, Pam, Morse, Barry, Peffer, Nancy, Beck, Heena, Lu, Jin, Cotty, Adam, Amegadzie, Bernard, Sweet, Ray
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Antibodies, Monoclonal - chemistry
Antibodies, Monoclonal - genetics
Antibody
Base Sequence
Cell Line
Coding sequence engineering
Codon
Codon engineering
DNA, Recombinant
Escherichia coli
Genetic Engineering
Humans
Molecular Sequence Data
Plasmids
Protein expression
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Summary: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.
ISSN:1046-5928
1096-0279
DOI:10.1016/j.pep.2007.05.017