Ivcd cell culture
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4Īs for any cell cultivation, also for perfusion processes the success depends highly on the available perfusion media. Despite high promises of this early technology, perfusion processes were rarely implemented in industry because of more complex process control, higher risk of process failure and higher media demands compared to traditional batch or established fed-batch processes. 1 Viable cell densities (VCDs) of up to 130 × 10 6 c/ml and high productivities, 2 as well as space–time yields (STYs) 3 were reported. Perfusion processes gained much attraction in the past for reliable production of labile therapeutic biologics including recombinant interferon-beta, coagulation factor VIII, recombinant follicle stimulating hormone, and different monoclonal antibodies (mAbs). This general workflow is universally applicable to any perfusion platform combining a specific cell line, basal medium, and established feed solutions. Obtained STYs of 0.4–1.2 g/L/d represent a 10-fold increase compared to batch cultures.
#IVCD CELL CULTURE VERIFICATION#
Two best performing novel media blends were finally applied to perfusion bioreactor verification runs to reach 200 × 10 6 c/ml within 2 weeks at minimum cell-specific perfusion rates as low as 10–30 pL/c/d. The final media formulation is supported by statistical response surface modeling of a set of cultivation experiments with blended media formulations. Then, the mixing ratio of selected feed supplements is optimized in small-scale semicontinuous perfusion cultures. First, the best performing feed supplements are selected in batch cultures. To develop clone-specific, high-performing perfusion media we present a straightforward and rapid two-step approach combining commercially available basal media and feed supplements using design-of-experiment. Economic processes rely on cultivation media supporting rapid growth in the exponential phase and high protein production in the stationary phase at minimal media consumption rates. This is due to increased space–time-yields (STYs) and a short residence time of the recombinant protein in the bioreactor. Perfusion cultivation of recombinant CHO cells is of substantial interest to the biopharmaceutical industry.