Orf virus (ORFV) is a promising vaccine vector candidate, capable of inducing a strong, long-lasting immune response without integrating into the host genome. Its broad host range and ability to express diverse antigens make it a valuable tool for recombinant vaccines and oncolytic therapies. However, effective analytical methods for characterizing ORFV materials, particularly for complex upstream samples, containing a mixture of viral particles, host cell components and process-related impurities, are limited. Additionally, traditional analytics often focus on detecting only one parameter at a time, requiring multiple assays to build a complete picture of viral production.
To address this gap, our study focused on integrating the liquid chromatography PATfix® analytical method as a multiparameter analytical tool to monitor and guide upstream process. The method allows fast and reliable profiling of both viral product and impurities, enabling insights into how process parameters, such as cell line, media, cell density at infection (CDAI), multiplicity of infection (MOI) and nuclease treatment influence virus production. Our approach emphasizes upstream optimization to enhance viral yield while reducing the downstream purification burden as well.
