CONICET | Buscador de Institutos y Recursos Humanos

Response Surface Methodology (RSM) is a collection of statistical and mathematical techniques useful for developing, improving and optimizing processes. The aim of this work was to analyze the influence of culture medium composition on growth and metabolism of a recombinant CHO cell line by means of this methodology. The response variables selected were: specific rate of growth (ì), specific rates of production of inhibitory metabolites, such as lactate and ammonium (qlac and qamm, respectively), and the time integrals of viable cells (IVC), lactate (TIlac) and ammonium (TIamm). Herein we specifically analyze the effect that different proportions of three hexoses exert on the cell growth and metabolism, as a preliminary step in the optimization of a culture medium which is being developed in our laboratory. To accomplish this goal, a constrained mixture experimental model was chosen and, in particular, a simplex lattice design was used. The analysis of the data was performed using the software Design Expert. In twelve simultaneous experiments, three pure blends (with replicates), three binary blends and four blends of three components were evaluated. The results indicated that, although similar specific rates of consumption and/or production were observed in some of the experimental units, different final maximal concentrations of cells or metabolites were achieved. These effects would depend on the specific metabolic pathways that the hexoses may transit within the cell and also on the relative proportions of each sugar in the culture medium. By assigning different factors of relative importance to every response of the system, it was possible to generate an improved desirability function which was used for selecting the best combination of these carbon sources which must be present in the medium, in order to optimize the growth and the metabolism of this cell line.

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