Comparative analysis of amino acid metabolism and transport in CHO variants with different levels of productivity.
Chinese hamster ovary (CHO) cells are widely used for the production of biopharmaceuticals; however, our understanding of several physiological elements that contribute to productivity is limited. One of these is amino acid transport and how its limitation and/or regulation might affect productivity. To further our understanding, we have examined the expression of 40 mammalian amino acid transporter genes during batch cultures of three CHO cell lines: a non-producer and two antibody-producing cell lines with different levels of productivity. In parallel, extracellular and intracellular levels of amino acids were quantified. The aim was to identify differences in gene regulation between cell lines and within culture. Our results show that three transporters associated with transport of taurine and β-alanine, acidic amino acids and branched chain amino acids, are highly upregulated in both antibody-producing cell lines but not in the non-producer. Additionally, genes associated with the transport of amino acids related to the glutathione pathway (alanine, cysteine, cystine, glycine, glutamate) were found to be highly upregulated during the stationary phase of cell culture, correlating well with literature data on the importance of the pathway. Our analysis highlights potential markers for cell line selection and targets for process optimization.
Centre for Process Systems Engineering, Department of Chemical Engineering, Imperial College London, London SW7 2AZ, UK.
Sarantos
S
Kyriakopoulos
Karen M
KM
Polizzi
Cleo
C
Kontoravdi
D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D000906:Antibodies / Q000235:genetics / Q000378:metabolism*; D001692:Biological Transport / Q000235:genetics; D016466:CHO Cells; D003412:Cricetulus; D005978:Glutathione / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000235:genetics; D013654:Taurine / Q000096:biosynthesis*; D015091:beta-Alanine / Q000096:biosynthesis*
D000596:Amino Acids; D000906:Antibodies; D026901:Membrane Transport Proteins; D015091:beta-Alanine; D013654:Taurine; D005978:Glutathione
Amino acid profiling; Amino acids transport; Biologics; Transcriptomics; qRTPCR