Model-based characterization of metabolism of the recombinant adeno-associated virus (rAAV) production via Human Embryonic Kidney (HEK293) cells

Abstract

In this study, we present a kinetic-metabolic model describing adeno-associated virus (AAV) production via Hek293 cells, encompassing 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 AAV (capsid and Rep proteins). For the modelling, Michaelis-Menten kinetic is 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 at low cell density culture, including concentration profiles of glutamine, glutamic acid, glucose, lactate, and ammonium, is utilized for fitting and computing the model parameters. The model resulting from the adjusted parameters well defines the experimental data. Subsequently, a Sobol-based global sensitivity analysis procedure is applied to determine the most sensitive parameters in the final model.