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| Table 1A: A CHO-GS production cell line was cultured in eight independently
controlled two liter mini-bioreactors (DasGip, CellFerm-pro). These bioreactors,
labeled R10H016 to R10H023 were set up according to Table 1, 1X being the
standard condition described below and in Materials and Methods. To determine
the impact of media components on productivity, a DOE study (three-quarterfactorial
“screening” design) using 25-2 resolution was implemented. With this
design, the effect of four media components, labeled Mach-1, F8, BRX and G8
was investigated. Each bioreactor received the indicated fraction of a standard
bioreactor condition. Standard bioreactor condition, defined as 1X for this study, is as follows: 806ml of growth media, which is a mixture of a proprietary chemically defined basal media termed Mach-1 supplemented with 8 g/L of a proprietary formulation of growth enhancers, termed F8, was seeded with 94 ml of inoculums (0.4×106 cells/ml), for a total volume of 0.9L. From D3 through D12, a proprietary formulation of vitamins and amino acids, termed BRX, was fed at a daily rate of 4% of the bioreactor volume. Additionally, from D3 through D17, a proprietary formulation of glucose and a select set of rapidly utilized amino acids, termed G8, were fed starting with 0.07% on D3 and ending with 1% on D17. |
