Use of a Plackett–Burman statistical design to determine the effect of selected amino acids on monoclonal antibody production in CHO cells

Abstract

Culture media design is central to the optimization of monoclonal antibody (mAb) production. Although general strategies do not currently exist for optimization of culture media, the combined use of statistical design and analysis of experiments and strategies based on simple material balances can facilitate culture media design. In this study, we evaluate the effect of selected amino acids on the growth rate and monoclonal antibody production of a Chinese hamster ovary DG-44 (CHO-DG44) cell line. These amino acids were selected based on their relative mass fraction in the specific mAb produced in this study, their consumption rate during bioreactor experiments, and also through a literature review. A Plackett-Burman statistical design was conducted to minimize the number of experiments needed to obtain statistically relevant information. The effect of this set of amino acids was evaluated during exponential cell culture (considering viable cell concentration and the specific growth rate as main output variables) and during the high cell-density stage (considering mAb final concentration and specific productivity as relevant output variables). For this particular cell line, leucine (Leu) and arginine (Arg) had the highest negative and positive effects on cell viability, respectively; Leu and threonine (Thr) had the highest negative effect on growth rate, and valine (Val) and Arg demonstrated the highest positive impact on mAb final concentration. Results suggest the pertinence of a two-stage strategy for amino acid supplementation, with a mixture optimized for cell growth and a different amino acid mixture for mAb production at high density.

Elucidating the effects of arginine and lysine on a monoclonal antibody C-terminal lysine variation in CHO cell cultures

“…Arg and Lys, two basic amino acids, have been well characterized by their noticeable roles in cell growth and mAb production during the cell culture process (Carrillo-Cocom et al 2014;Gonzalez-Leal et al 2011). They were also found to stabilize mAbs during the purification and formulation process (Falconer et al 2011).…”

5.3 Nutrient Media for Cell Culture Technology

“…This procedure results in a minimisation of costs and analysis time. Its use for the analysis of the effect of various compounds and supplements for growing different cell lines has proven to be very suitable [38,39,16].…”

Fermentanomics: Relating quality attributes of a monoclonal antibody to cell culture process variables and raw materials using multivariate data analysis

“…Researchers have attempted to improve cell lines, media composition, and bioreactor conditions to achieve these objectives . Alvarez and coworkers have reported a two‐stage strategy for supplementation with different aminoacid mixtures optimized for cell growth and mAb production. In another study, Geaugey etal .…”