Model-Based Characterization of the Metabolism of Recombinant Adeno-Associated Virus (rAAV) Production via Human Embryonic Kidney (HEK293) Cells

In this paper, we present a kinetic-metabolic model describing adeno-associated virus (AAV) production via HEK293 cells that encompasses the main metabolic pathways, namely, glycolysis, tricarboxylic acid cycle (TCA), pyruvate fates, the pentose phosphate pathway, anaplerotic reaction, amino acid metabolism, nucleotides synthesis, biomass synthesis, and the metabolic pathways of protein synthesis of the AAV (capsid and Rep proteins). For the modeling, Michaelis-Menten kinetics was assumed to define the metabolic model. A dataset from bioreactor cultures containing metabolite profiles of adeno-associated virus 6 (AAV6) production via triple transient transfection in a low-cell-density culture, including the concentration profiles of glutamine, glutamic acid, glucose, lactate, and ammonium, was utilized for fitting and computing the model parameters. The model that resulted from the adjusted parameters defined the experimental data well. Subsequently, a Sobol-based global sensitivity analysis procedure was applied to determine the most sensitive parameters in the final model.