Formulation of a culture medium for mammalian cells used in recombinant protein production

DIDIER C, ETCHEVERRYGARAY M, KRATJE R, GOICOECHEA H

Pinamar, Argentina

Congreso; XLI Annual Meeting, SAIB & X Panamerican Association for Biochemistry and Molecular Biology Congreso.; 2005

SAIB

In this work the effect of a mixture of three hexoses (H1, H2 and H3) is evaluated, combined with a mixture of three energy-provider compounds (E1, E2 and E3), to determine their optimal proportion in a medium being developed for culturing recombinant mammalian cells. The effect of hexoses was analyzed using an augmented simplex lattice {3,2} design, while a constrained {3,2} simplex-centroid design was chosen for the mixture of E compounds. These experimental designs are combined in a crossed mixture, giving a total of 63 experiments. The empirical data were used to train an artificial neural network (ANN) for each response. The ANNs were then used to predict the responses for 300 different combinations of the six compounds, and based on this information a partial desirability function (di) was generated for every response. The di were then combined in a global desirability function D, and the blend for which it is predicted to obtain the highest value of D was selected. This combination was a mixture of H3=20.33 mM, H2=2.33 mM, H1=2.33 mM, E1=1.000 mM, E2=0.775 mM and E3=3.325 mM. Remarkably, H3 is not frequently found in commercial media.