Evaluation of Sodium Acetate in HEK293 cells producing the ECD-HER1. Metabolomics

Description

Our research was focused to the evaluation of Sodium Acetate (NaAc) in the HEK293 cell culture. HEK293 is employed for the stable production of the extracellular domain HER1 at the Center of Molecular Immunology, a glycoprotein that is one of the extracellular domains of the epidermal growth factor receptor. We study the effect of NaAc over the glycoprotein production, but also we search its impact over metabolism. The data presented correspond to the HEK293 cell culture in batch mode, using concentrations of NaAc from 0mM; 4mM; 6mM; 8mM; 10mM and 20 mM. Samples were taken daily from the culture, in order to measure cell density by using Trypan blue dye exclusion, protein concentration by a homemade ELISA sandwich, and metabolites by derivatization method and Liquid Chromatography-Mass Spectrometry. After obtaining the corresponding measurement of each variable, we calculate the external specific rates by using mass balance equations, such as specific growth rate, specific protein production rate and specific metabolites uptake/production rate. Also, it was represented the profiles of the measured variables across the culture time, with the aim of identifying the growing phases. On the other side, the use of Metabolic Flux Analysis (MFA) was employed to predict the internal metabolic flux of this modified HEK293, with and without NaAc. To carried out MFA method, it was created a Matrix containing the concentration of 19 metabolites including aminoacids, Glucose and Lactate. To do that, programming codes were created, and metabolic models were adapted considering reactions and metabolites that are presented not only in the Recon2 network, but also were added reactions and metabolites that are connected with the metabolism of acetate. The data presented, and the obtaining results, show an increment in protein open a new paradigm for the scientific community, in particular for those who are interested in the metabolism of HEK293 cells.

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