Optimization of multiplicity of infection and serum concentration to improve the titer of adenovirus by "design of experiments"

Abstract Objective: To improve the adenovirus titer by optimizing the existing adenovirus-dependent HEK293 host cell expansion and replication experiments through a “design of experiments” (DoE) approach. Methods: Various concentrations of host HEK293 cells were infected with a different multiplicity of infection multiplicity of infection(MOI) of adenovirus using optimal experimental conditions, and the titers were detected using an adenovirus titer detection kit to evaluate the optimization effect. Results: Using DoE, we found that a serum concentration of 2.2%-2.4% and a MOI of 2.3-2.5 could maximize the adenovirus titers. Conclusion: Optimization of the experimental system of adenovirus culture using DoE could significantly shorten the production time and improve the titer of adenoviruses, providing an experimental basis for the current study and the subsequent expansion of production experiments.