Native elution immunoaffinity chromatography for the efficient purification of vesicular stomatitis virus

Vesicular stomatitis virus (VSV) is a promising platform for oncolytic virotherapy and vaccine development. However, its purification remains challenging due to the sensitivity of enveloped viruses to conventional downstream processing conditions. In this study, we investigated the transferability of our previously developed native elution immunoaffinity chromatography (IAC) concept to efficient VSV purification and systematically evaluated the key factors governing its performance while preserving virus integrity and infectivity. Polyclonal anti-VSV IgG antibodies of guinea pig and human origin were immobilized on hydrazide-activated monolithic columns and evaluated for virus capture and recovery. Although both columns exhibited high binding capacity, successful native elution was achieved only with human anti-VSV antibodies, which, in contrast to the guinea pig anti-VSV antibodies, exhibited substantially lower specific neutralization potency. Elution was performed under physiological conditions using high-molarity amino acid solutions. Among the tested eluents, arginine was found to be the most efficient, enabling high recovery of total and infectious virus particles. Process analysis revealed near-complete removal of host cell DNA following benzonase treatment, whereas a fraction of host cell proteins co-eluted with virus particles. Further experiments suggested that these proteins were primarily associated with particulate material rather than being present as soluble impurities. These results demonstrate that native elution IAC using arginine enables efficient purification of VSV under mild conditions, achieving high recovery of infectious virus and providing a strong foundation for developing scalable purification processes for enveloped viral therapeutics.