Oncolytic measles viruses produced at different scales under serum-free conditions

Measles virus (MV) with attenuated pathogenicity has potential as oncolytic agent. However, the clinical translation of this therapy concept has one major hurdle: the production of sufficient amounts of infectious oncolytic MV particles. The current study describes oncolytic MV production in Vero cells grown on microcarrier using serum-free medium. The impact of the number of harvests, cell concentration at infection (CCI), multiplicity of infection (MOI), and temperature on MV production was determined in different production scales/systems (static T-flasks, dynamic spinner, and bioreactor system) and modes (batch, repeated-batch, and perfusion). Cell growth, metabolic, and production kinetics were analyzed. It was found that the number of harvests had the strongest positive impact on MV yield in each production scale, and that high temperatures affected MV yield adversely. Moderate MV titers were produced in T- and spinner flasks at 37 degrees C (approximate to 10(7) TCID50 mL(-1), where TCID50 is tissue culture infective doses 50%), but stirred tank reactor (STR) MV production at 37 degrees C yielded up to 10000-fold lower MV titers. In contrast, at lower temperatures (32 degrees C, 27 degrees C), 1.4 x 10(7) TCID50 mL(-1) were achieved in the STR. Variations in MOI and CCI had almost no influence on MV production yield. The current study improves oncolytic MV production process understanding and identifies process bottlenecks for large-scale production.