Abstract
Oligosaccharides are generally considered to be un-utilized for growth of mammalian cells because their permeability across the cell membrane is low. However, in our previous study, we discovered that CHO and HEK293 cells consume maltose in culture media without serum and glucose. This is interesting because the transporter for maltose in mammalian cells has not been discovered to-date, and the only animal disaccharide transporter that is recently discovered is a sucrose transporter. The application of oligosaccharides in mammalian cell-based biopharmaceutical manufacturing can be beneficial, because it can theoretically increase carbohydrate content of the culture medium and decrease lactate production. Here, we first determined that specific maltose consumption rate in CHO cells was similar to galactose and fructose at 0.257 ng/cell/day. We then demonstrated that CHO cells can be cultivated with reasonable cell growth using higher maltose concentrations. After which, we evaluated the use of maltose supplementation in the production of a recombinant monoclonal antibody in batch and fed-batch cultures, demonstrating improvements in recombinant monoclonal antibody titer of 15% and 23% respectively. Finally, glycosylation profiles of the antibodies were analyzed.
Highlights
Mammalian cell cultures are commonly used for the manufacturing of recombinant glycoprotein products because of its ability to fold complex proteins and to add human-like glycans to glycoproteins
We proposed that the specific maltose consumption rates from SH87 maltose supplemented batch cultures can be fitted in a Monod model to obtain a maximum specific maltose consumption rate of 0.257 ng/cell/day and an affinity constant (Ks) of 7.03 g/l
The model is validated using maltose-adapted CHO-K1 cells cultured without glucose, serum or protein supplement, to a viable cell density (VCD) greater than 106 cells/ml
Summary
Mammalian cell cultures are commonly used for the manufacturing of recombinant glycoprotein products because of its ability to fold complex proteins and to add human-like glycans to glycoproteins. We reported our discovery that mammalian cells consume maltose, a disaccharide, in serum-free cell culture: we demonstrated that CHO and HEK293 cells can be adapted to serum-free culture media containing maltose as the sole carbohydrate source, intracellular maltose was detected in maltose-supplemented cell culture, and that maltose in a protein-free chemically-defined medium is only depleted in the presence of cells[11]. This is somewhat surprising because currently, the only animal disaccharide transporter that has been reported is a sucrose transporter in Drosophila melanogaster[12]
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