title: Using simple models to describe the kinetics of growth, glucose consumption, and monoclonal antibody formation in naive and infliximab producer CHO cells.
abstract: Despite their practical and commercial relevance, there are few reports on the kinetics of growth and production of Chinese hamster ovary (CHO) cells-the most frequently used host for the industrial production of therapeutic proteins. We characterize the kinetics of cell growth, substrate consumption, and product formation in naive and monoclonal antibody (mAb) producing recombinant CHO cells. Culture experiments were performed in 125 mL shake flasks on commercial culture medium (CD Opti CHO™ Invitrogen, Carlsbad, CA, USA) diluted to different glucose concentrations (1.2-4.8 g/L). The time evolution of cell, glucose, lactic acid concentration and monoclonal antibody concentrations was monitored on a daily basis for mAb-producing cultures and their naive counterparts. The time series were differentiated to calculate the corresponding kinetic rates (rx = d[X]/dt; rs = d[S]/dt; rp = d[mAb]/dt). Results showed that these cell lines could be modeled by Monod-like kinetics if a threshold substrate concentration value of [S]t = 0.58 g/L (for recombinant cells) and [S]t = 0.96 g/L (for naïve cells), below which growth is not observed, was considered. A set of values for μmax, and Ks was determined for naive and recombinant cell cultures cultured at 33 and 37 °C. The yield coefficient (Yx/s) was observed to be a function of substrate concentration, with values in the range of 0.27-1.08 × 10(7) cell/mL and 0.72-2.79 × 10(6) cells/mL for naive and recombinant cultures, respectively. The kinetics of mAb production can be described by a Luedeking-Piret model (d[mAb]/dt = αd[X]/dt + β[X]) with values of α = 7.65 × 10(-7) µg/cell and β = 7.68 × 10(-8) µg/cell/h for cultures conducted in batch-agitated flasks and batch and instrumented bioreactors operated in batch and fed-batch mode.
authors.affiliation: Centro de Biotecnología-FEMSA, Tecnológico de Monterrey at Monterrey, Ave. Eugenio Garza Sada 2501 Sur, C.P. 64849, Monterrey, Nuevo León, Mexico.
authors.forename: Julián
authors.initials: J
authors.lastname: López-Meza
authors.affiliation: Centro de Biotecnología-FEMSA, Tecnológico de Monterrey at Monterrey, Ave. Eugenio Garza Sada 2501 Sur, C.P. 64849, Monterrey, Nuevo León, Mexico.
authors.forename: Diana
authors.initials: D
authors.lastname: Araíz-Hernández
authors.affiliation: Facultad de Ingeniería Química, Universidad Autónoma de Yucatán, Periférico Norte kilómetro 33.5, C.P. 97203, Mérida, Yucatán, Mexico.
authors.forename: Leydi Maribel
authors.initials: LM
authors.lastname: Carrillo-Cocom
authors.affiliation: Centro de Biotecnología-FEMSA, Tecnológico de Monterrey at Monterrey, Ave. Eugenio Garza Sada 2501 Sur, C.P. 64849, Monterrey, Nuevo León, Mexico.
authors.forename: Felipe
authors.initials: F
authors.lastname: López-Pacheco
authors.affiliation: Centro de Biotecnología-FEMSA, Tecnológico de Monterrey at Monterrey, Ave. Eugenio Garza Sada 2501 Sur, C.P. 64849, Monterrey, Nuevo León, Mexico.
authors.forename: María Del Refugio
authors.initials: Mdel R
authors.lastname: Rocha-Pizaña
authors.affiliation: Centro de Biotecnología-FEMSA, Tecnológico de Monterrey at Monterrey, Ave. Eugenio Garza Sada 2501 Sur, C.P. 64849, Monterrey, Nuevo León, Mexico. Mario.Alvarez@itesm.mx.|Biomaterials Innovation Research Center, Division of Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, 02139, USA. Mario.Alvarez@itesm.mx.|Harvard-Massachusetts Institute of Technology Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA. Mario.Alvarez@itesm.mx.
authors.forename: Mario Moisés
authors.initials: MM
authors.lastname: Alvarez
keywords: Biopharmaceuticals; CHO; Kinetics; Luedeking–Piret; Monod; mAb