Parameters affecting dengue virus replication in Vero cells

Dengue has become a global threat affecting millions of people annually and contributing to a significant mortality burden. Although vaccination is the most effective approach to prevent this disease in dengue-endemic regions, the development of a robust and scalable production process is urgently required to maximize vaccine yield. The current study aimed to optimize the key growth parameters of four recombinant live attenuated dengue serotypes, including harvest schedule, temperature, Multiplicity of infection (MOI), and metabolite concentration. Growth kinetics data, followed by a scale-up study, established the feasibility of multiple harvests for all four serotypes within a defined production window, with the first harvest optimally occurring between Days 4 and 8. All serotypes demonstrated strong responsiveness to low MOIs and temperature shifts, resulting in 2- to 25-fold increases in titer within a reduced time frame. Furthermore, metabolite profiling revealed distinct glutamine and glucose consumption dynamics, highlighting its importance for optimizing infection media. Taken together, these findings provide valuable insights into dengue virus production and define critical parameters that can support future optimization studies and large-scale production.